AU2016216524A1 - Antibodies that bind human CD27 and uses thereof - Google Patents

Abstract

Isolated monoclonal antibodies which bind to human CD27 and related antibody-based compositions and molecules are disclosed. Also disclosed are therapeutic and diagnostic methods for using the antibodies.

Description

ANTIBODIES THAT BIND HUMAN CD27 AND USES THEREOF

The present application is a divisional application of Australian Application No. 2013203270, which is incorporated in its entirety herein by reference.

Background of the Invention

Interactions between T cells and antigen-presenting cells involve a variety of accessory molecules that facilitate in the generation of an immune response. One such molecule is CD27, which binds CD70 and belongs to the tumor necrosis factor receptor (TNF-R) superfamily (RanheimEA, et al., Blood. 1995 June 15;85(12):3556-65). CD27 typically exists as a glycosylated, type I transmembrane protein, frequently in the form of homodimers with a disulfide bridge linking the two monomers. The disulfide bridge is in the extracellular domain close to the membrane (Camerini et al., J Immunol. 147:3165-69 (1991). CD27 may also be expressed in a soluble form (see, e.g., van Oers MH, et al., Blood. 1993 Dec 1 ;82(11):3430-6 and Loenen WA, et al., Eur. J. Immunol. 22:447, 1992). Cross-linking the CD27 antigen on T cells provides a costimulatory signal that, in concert with T-cell receptor crosslinking, can induce T-cell proliferation and cellular immune activation. CD27 is expressed on mature thymocytes, on most CD4+ and CD8+ peripheral blood T cells, natural killer cells and B cells (Kobata T, et al., Proc. Natl. Acad.

Sci. USA. 1995 Nov 21 ;92(24): 11249-53). CD27 is also highly expressed on B cell non-Hodgkin’s lymphomas and B cell chronic lymphocytic leukemias (Ranheim EA, et al.,

Blood. 1995 Jun 15;85(12):3556-65). Additionally, increased levels of soluble CD27 protein have been identified in sera or sites of disease activity in parasitic infection, cytomegalovirus (CMV) infection, sarcoidosis, multiple sclerosis, and B-cell chronic lymphocytic leukemia (Loenen WA, et al., Eur. J. Immunol. 22:447, 1992).

Agonistic monoclonal antibodies against CD27 have recently been shown to promote T cell responses and show promise as anti-cancer therapeutics (see e.g., Sakanishi T, et al., Biochem Biophys. Res. Commun. 2010 Feb 18 and WO 2008/051424). However, while the results obtained to date establish CD27 as a useful target for immunotherapy, it is unknown which particular features of anti-CD27 monoclonal antibodies are especially advantageous for therapeutic purposes. As such, there is a need in the art for further insight into the specific functional properties that make anti-CD27 antibodies therapeutically effective, as well as improved therapeutic antibodies against CD27 which are more effective for treating and/or preventing diseases. SMiaEBmiiMMmiim

The present invention provides ouer alia isolated anti-CD?.7 antibodies basing purdeuiar functional properties which cun be holed with advantageous and desirable therapeutic effects. SpeeiileaHy: ami-ClXC monoclonal antibodies capable of up-vegtdaiiag Tcdt mediated immune responds use., as evidenced by ind aeon east or enhancemem of anriaeo-specitic T eels responses*, which are particularly· well-suited tor combination wuh vaccine therapies, base been generated and character! red by way of the present invention. hi one embodiment. agonist an si'CD.'.’7 andbudies can ess ion see the immune wspouse against cancels os' infectious diseases by combination with acme s aeetnabon, or by enhancing endogenous immune iv-ponsc1- Such antibodies sn.ay also di techy or indneetiy mdace eytokkn· expression Additionally, anii-Cl>2? antibodies that down-regnhte T ceil mediated immune responses, whteh ;uv particularly veil-suited tor treating immune disordetsysueh as graftfejeetidn, allergy and gufoi'snpreoo diseases^ have been, generated and charaeteseerh Still Sunher, ant· CD2? sniiiovde-. mat inhibit growth -.a"Cl >77 expressing ceils by direct cell hilling mechanism*·· u g., antibody dependent cell-mediated csunoetciis (ADCO) and/or complement dependent celltdarcytotoxicity iCDOO). which are particularly effective in uvaony a wide variety ot diseases involving cell proliferation <cancersd have been generated and characterized. krone embodiment, the antt-CD27 antibodies oi the present invention exhibit one or more ot the following properties: (a): blocks hidiom of sCD70 to. CDS? by at least about 711% at an andbadg cdacentmtion of 10 pg/mi; ibt binds to human CD27 svitb an equilibrium dissociation constant Kd ot' jO'" M m less, or alternatively an equilibrium association constant. Ka of l(f ’ M‘l or greater; (ci induces specific complement mediated cytotoxicity (CDO of CP27 expressing cells of at least iOD· at an antibody eoncentnaion of 3 pg/ml and approximately $%. rabbit serunt complement:: (d; induces antibody dependent celi -meditued cytotoxicity {ADCO .specific lysis of OD27 expressing cells of at least. 1071 at an antibody eoncenuation of a pg/mt and ratio ot effector, target ceils of 7ό; 1 ic> improves median survival by in least 20'Τ trt severe combined xmmitnodeftdeney -(SC'IB) mice peat tbmor cell mbaulalitm ik viv&amp; ($ x 16'’ -Raji. celts or f: n. 10*' Datidi cells? when admin?srered at 0.3sng ft.p. ! at least twice a week tor3 weeks compared to mice to whtch antibody is not adnnnkiesvib <f? induces or enhance amigen-spccblc immune responses ot combination with a vaccine or endogenous antigen: ies induces or enhances antigen-specific ΊΊ11 stmim:tieres|Mtises::tB cspiltittatlon with a vaccine or endogenous antigen; (h) induces or enhances antigen-specific T~edl proliteralton or activation in combi nation with a vaccine or endogenous antigen: (t) reduces or inhibits T-ceil proliferation or activation: ijt induces or enhances T-ceil activity : separate or sequential TGB activation; tk} blocks binding of sCD?0 to CD2? by at teas? about ?0vf at an antibody couamruration of 10 μα/mi and ; educes or inhibits T-ceil activity when not capable of binding to. or having reduces! binding to Ft: receptors: {) i results ·η loss than bO'T depletion oi CDJ-t-T-celis (miter than NK cells? in macaques when administered at 3 mg/k g : i v.) over the peiiod of 2d days immediately fwllowing administration: or (mi icsuHs in less than 30'2 depletion t>f memory B-cells in macaqu&amp;s when administered at ? mg/kg i i.v ? over the perk'd of 2d day-, immediately following administration. . in a particular embodiment the «tlborfies of the mvemi om fnneti ot tal properti e s.

Accoidinaly tn one aspect, the insentient provides ontbCD27 antibodies that indtice and/or enhance an immune tespouse ti .y.. a '1 cell mediated immune respon-'e!. In a further embodiment. the antibodies inhibit the binding of CD70 to OD7.7 on cells. Particular antibodies has ing toe-m combinations of properties include mAh ! t o comprising heavy and/or light chain saiiabie regions sequences comprising SEQ ID NOs: 37 and/or -13. respectively; Alternatively. the antibodies do not inhibit the binding of (/1370 to C027 on cells Particular antibodies having these combinations of properties include mAh 3H8 comp-rising heavy und/or light chain variable regions sequences comprising S1.I.Q ID NOs: 7 imd/or 13 owpecnvdy , o; 7and/or 19. respectively i. Such antiCDa/ antiboebe-.: also cun be jinked to a eevosul molecule- (c.g. as a h;specific molecule) having a binding spec· deity w hich is ddferetu from the anybody. such as n 1' edi receptor tc.g,, C:D3. CD25. CD 13 n •CD154J. or an Fc receptor u FcfKI tCD64). FcyROA (CD32L FcvRUBi (CD32;. FcyRMB2 (CD32). RyRUtA (CDI6a>. FcyRHIB <CDl6bj, Fcr.RI, FcuRff (CD23l FcaRI {CDi>5’\ Fcu/pR. and FeRu). or an NK teecpior (>\g. CD56), or a B cell receptor (e.g. CD 19, cd;; ;

AmiD-dies intended to he used for induction or enhancement oi immune ;espouses according to the: present invention may have a functional Fa don tain permitting bmdme to Fe receptors. and oner include a -nutated re domain having laciva^ed levels of binding to Fc receptors. in anoihcs aspen. the invention provides α·πο(.Τ>27 antibodies that, dovvn-regolaie Teell mediated torn tune re peon see by inhibiting she binding of' CD.?? toCD/O on cells which express these proteins fn a particular embodiment. the an tamales inhibit Use binding of' soluble CD70 -:-:00701 to (“027 expressing cells by at least about 7()¾. Particular antibodies Etlhng within this class include. c.y.. mAh comprising heavy and/or light chain variable regions sequences comprising SCO F? NOx537 atsd/ordo tmAb IF’5). SEQ (D NOs: C) and/or 55 (mAh I HHh or SEQ ID NOs; 103 and/or J 09 (mAh 3H1

In yet another aspect, the invention provides' $mi*CD27 antibodies that induce Or etfhance eifeetor eel! function (e, g,, cell hilling v.i A either ADDC and/or CDCr In .one embodiment, the antibody induces ;.tt. least about 30((- specific lysis of CD27 eNpressimt ceIN via A DOC at an antibody concent! ation ot’ iOpg/ml and/or induces at least about 3o< rODC ot OD2? expressing cells at a concentration of 10 pg/ml. Particular anhbodie-. falling within ibis class exhibiting ADCC ei'i’eeior 1 unction include, e.·'.. ;c..e.. mAh eompnvma heavy and/or light chain ·- arluble region sequence1- comprising SEQ ID NO;;: 61 and/or 6? tmAb I CD;. SPQ ID NOs: 85 and/or 91,85 and/or 97 (rnAb 3A10t. SEQ ID NOs:37 and/or 43 fmAb iF5f, SF.Q ID NOs: 7 and/or 13.7 and/m is) «mAh 3118;.. SF.Q ID NOs; 49 and/or 55mtAb IH8). or SEQ ID NOs: 103 and/or 109 (mAh 11 1 12). In a lurlhe- embodiment the antibody also inhibits binding of (7070 to CD27 on coils. Particular antibodies having these eonminadons of tuned mis include, e,g„ iV,y,. ntAb comprising heavy and/or light chain variable regions sequences comprising SF.Q ID NOs; 37 atul/or 43(mAb IP3;-, SEQ If) NO·.·: -Iv and/or 55 {mAh 1118;. SEQ IE> NOs. 105 and/or 109 mAh 3H 1.2). AhemahveN. the antibody .induces .ADCC and/or CTX? ns described above, hut does not inhibit binding ofCD70 to (’4027 on cell·-· Particular antibodies having these features include, e y . ns Ah comprising heavy and/or light chain variable regions sequences euro prising SF.Q ΙΠ NOs: (>) and/or 67 (mAh iGh), SEQ 0.) NOs;85 and/or 9L 8S and/or 9? tm.Ab 3ΛΙ0), SEQ ID NOs;7 and/or 1,7. 7 and/or 19 (mAh 21 IS).

Ami-Cf>2.7 ant ib; iiiii.'s capable of inducing or enhancing c dec ίο; eel! function y.. ADCCV and/or COC; can also he constructed to include an Fc region oh-eh suitably contribute·' a binding specificity lorn spechie Fc recepstir urg’., FcyRl ;CT>6d K EcyRILA (CD32).

FcyRIlBl (ΓΠ32), FcyRlJB2 KTGT. FcyRfHA (CD!6a). PcyRIIIB (CDibb), FecRF FceRM ¢(71.)23). FcuRI (CDST. Fcn/pR. and FcRni,

In a iWther embodiment there -> proxided a method lor enhancing an immune ; espouse agamst an suticcn in a subject in need ths-neof Fy, ailmlniMcnug to the s urn ecu ρ an ami-CHi!? am Tody and Hi an antigen, o Pet too the anu (TO" antibody «'s adm · n i mo; m i separately iron; and before the antigen o adtriinraered.

Typically in Midi a method the atm 0)27 amd body αίαν he administered between at least 2 and 96 hours before th·,.· antigen. For evtnmie. tn such a sneihosi the ami-CDm antibody rnay be administered at learn 2 horns hetoto the as;;seen lor example at leas t i 2 hoars before the antigen. Mutably at least IM hours ludV-se the antigen, at least 48 hours before the. hhhgen or atlMsI 72 hours feefore the antigen, wherein the IT FI agonist is a TLRo agonist.

In a further embodiment there Is provided a method im enhancing an immune response against an antigen in a subject in need thereoFbs shnuitanonsly. separately or §ί.»ψαγ;η daily administering to the subject: 1) an ann-CD27 antibody: ii t a IT FI agonist' and iii) optionally,, the. an 4 gen..

In a preferred embodiment of such a method the TLR agonist is a TL;R3 •agonist, for example hut not limited to Poly SO: EC. (7027 expressing ceils include arts and all cells that express; CP27. including, hut not limited to B ceils, NK cells and Ί ceils. In a particular embodiment, the C7D7.7 expressing cell·; include cancer cell lines such as Jerkai ceils. Rati cells. Ramos cells ami Daudi cells, hi another embodiment, the CD27 expressing cells are tumor cells or entice· cells, in another embodiment, CD37 espresGng cells include B cell1;, NK cells, and T cells intending T cell·: that are found infiluating mrnots, also calk'd tumor infiltrating lymphocytes.

Parttcuiar antibodies of the invention comprise beam and light chain variable regions' that util «re particular human gemttiues. os,, arc encoded by the gems I me genes, but include genetic maoangemonl·: and nunations, mm. somatic mutation;, v- Inch occur during antibody maturation. in otic embodiment, the heavy chain variable region of the antibodies >.>! the present mvmnlon is derived horn a human ©cnnl'me 3 7 or 3-33 sene, in another embodiment, the hem chain variable region of the antibody D derived irons a human gormbne 3-20, 3- 11.2-1 I D-lb, or 1-13 gene. In a particular embodiment, the heavy chain variable region of She antibody m der ived from a human germfme V5! 3-7 or Vi{ 3-33 sene and the hah? chant triable reason ot the antibody l·- derived frotn a human germ line 303-20. VK Λ-1 1. VK ID-lb. o? \ -. I 13 gene. A..3¾3-33 Accession NcAaP44382) as tollows:: ! vqNesgggs vqpgmhls causal-Dt ygmhvvvrqap gkglam van oidesmyya M User:; sins rdssri-.TyI mm.-homl? avmcnk tSEO ID NO; 3; A \% 3-7 gerrnllae.se{|tienee N provided fOortbaak Accession No ΑΑΡ443Β9|'^·$^|%ί<»ν^ί i vqivesgggl vqpggslrD eaasgftfsn symtxvv rqap gkglew', am Updgxdknyl 0i nsvrgrnia rdmmlmsyl qnthsfraedt aivsevt ?SHQ ID NO; 4;

In another mnbodinsc'iu. -he heavy Chain sartahlc region 03R3 sequence 1-, • elected bvrn die croup cerum-mne m SLQ II > V, >m Ml 78 Ur. 5?. 04. ~N kg IUP. and conservative sequence modhienboir- thereof m\y.. eon«er\.atj\e ammo acid suhmmnmnsU j he nmiuOle- may fn nor: include a light chain amiable region (OR 3 sequence selected irons tile stoop enrooting of\LQ ID SO1' k\ 77. 3 t. by. gx Ό. 87. i!-k 100, 11 2. and conservative -equeace mojnlmumns u?mm.N in another viotxxhvnem the heavy vrasi; CDR2 and i '1 H< t sequences are selected from 3h0 ID N'Os. v 0.13'?. 51.6 <, "5, ST. 105. and SEQ ID \<)v S do. 38. pit a;:. Ό kb. Ibid, respoc?:'. cU. and mmsew arise sequence modifications thereof. The light chain CDE2 and C13RI sequences ore selected from SEQ ID NOs; 15, 71. 33. 45 h 7, 69. B1.03. 90. ill. and SPQ ID Νί )s: 14, .20, 32,44, 56, f.N, BO, 02. 03, 1! 0. nsp»ctive]Y. :utd cause· votive sequence modifications thereof, in ..still another embodiment, the invention provide;? an isolated antibody That binds CD:27 and includes heavy and light chain variable region CDRI»CDR2 and 0DR3 seqe&amp;nces: «.elected from the group consisting of; ft) a heavy chain 'variable region CDR 1 comprising SEQ ID NO; 38; a heavy chain variable region CDR2 comprising SEQ ID NO; 39; a heavy chain variable res son CDR3 mtmiprmnm SF.Q 01.) NO: -Hr, a light chaimva.riahle region CDRI comprising SEQ ID NO: 44; a baht chuui Variable region CDR2 comprising SEQ ID NO: 45. a tight chain variable region CDR8 compriama SEQ ID NO; 40: or con\enative sequence modifications Thereof; f;ii> a heavy chain variable region CDRI con-prising .SEQ ID NO: 50; a heavy chain variable region CDR2 comprising SEQ ID NO; M; a heavy chan- variable region CDR3 comprising SEQ I'D NO: 52: a Rah; chain variable region CDRi comprising, SEQ ID NO: 56; a light chain variable region CDR2 comprising SEQ ID NO: >7; a light chain variable region CDR2 comprising SEQ ID NO: 58: at converv alive sequence modificanonv thereof; (ill) a heavy chain variable region ODRI comprising SEQ ID NO: 104; a heavy chain variable reston CDR2 t ompnsiug SEQ ID NO: K;5, a heavy chain variable region ('DR.5 comprising SEQ 10 NO; 10b' a light chain variable region CDRI eompn-eng SEQ ID NO: I lb; a right chain variable region C DR a comprising SEQ ID NO: III. a lighi chain variable region ODR8 comprising SRQ ID NO: i 12: or conservative sequence mudificafions thereof: (¾) a heavy chain variable -egienCDRl comprising SEQ ID NO: 86' a heavy chain variable region ODR2 comprising SEQ 111) NO: 87: heavy chain van able region ODR5 comprising SEQ ID NO; bib v light chain variable region CDRI comprising SEQ ID NO: 92 or 98; a light chain variable region ('DR2 comprising SEQ ID NO: 98 or 99: a light chain variable mgion CDR3 comprising SEQ ID NO: 94 or 100; or :. misers alive sequence modhleahous thereof: (v) arheavy chain variable region ODRI comprising SEQ ID NO; 26; a heavy chain variable region CDR2 comprising SEQ ID NO: 27; a heavy ·. ham s-triable regime E'DRu- comprising SEQ ID NO: 28; a light chain \unabE- teglo-a CDRI cornprisiug SEQ ID NO; 32; a ugh: chain variable region QDR2 ecurq'iriring SEQ NO: 33; a light chain variable region OPR) c> ηοηποην M .Q (0 NO: 34. or conservative .sequence m edifications thereof: icl) a hoifvf eligfa x arlahle i'egiou CDR i compriMna SEQ ID NO: 74; a heavy chain variable region QDPkl comprising SEQ ID NO; 70 a. heavy chain van a hie region CDR3 comprising' SEQ ID NO: 7b;

a light chain variable region 0DR1 comprising SEQ ID NO: SO: a light chain variable region CPR'2. coin prising SEQ ID NO: hi: a hglu chain variable region CUR3 comprising SEQ ID NO; 83: or conservative sequence modiikaUons itereoQ (x us a bear <, chain vartaNe uqm.'rt (Ί )R i enrupnnng M Q ID NO; S' a he.sxy chain xaiiahlv tegaon CDR2 composing Si.<3 IP NO. 3. a hens y chain vrnaabie reaion C PR3 *. ompnsing SEQ if) NO' kb ;i light chain \ unable region ODRI composing SEQ ID NO. i4 or 70' a light chasn van abb' ieg;on i Dr.3 compming SEQ U) NO; ; ^ or 2 I: a be;·; cb.no mu able region fT)R3 C'-nnuieUig Si Q ID NO: 16 or 33; or λ onsers.thxe sequence modification-. these* if- and

Qrifi) a heavy chain variable region C'DR I conspriying SEQ ID NO: 62; a heavy chain variable if»ionCDR3 comprising SEQ ID NO; 63: a heax y chain variable region CDR3 comprising SEQ ID NO: 6-1: a light chain xariable region CDE i comprising SEQ ID NO: OS: .a light chain 'variable region CDR2 comprising SEQ ID NO: 60: a light drain variable region CPR3 comprising SEQ ID NO: 70; or .confervuttvc sequence modifications thereof h; another embodiment. the be a vs chant \ enable region QDR3 sequence comprises an ammo auci sequence ^elected to>:a tlte con-va-ms -a quern, e' R :(3..73.): tS.L.O, ) :(,:.I .T.VR -»QE-ΝΓ 11 - ) ;V.r..-· (Nl.P.m ;tl.V.- * <R. -: lO.M. i (Oil L. 1 AN') ik.G.NkN riJ.F.V.Y: if .Id iD.l'O ilLLLD ) tSEQ ID NO : i D, \λ herein '·“ drumes the opt;eat ol no ammo acid re mine being present at that con -earns pm-mem The arm bodies nm> Umber include a light chat n variable region CDR3 sequence comprising an ammo acid sequence selected from the consensus sequence: O iF.R.Y1 tN.Si qX.I.S; t V \V t P R,R: T tSIli) ID MU: 1 14). wherein denotes the option of n<> amino acid tv-idae being ptessmt at that consensus poeitiC·!:. in a;50the; embodiment. the boas y >..ham variable region {'DR 2 sequence comprises ati anti no acid sequence -.elec-red from the consensus sequence: i (K.W; t Y.N.Qt D G S «Ε.Ν'» i'K\Qi iSEQ ID NO: I 10), wherein "-''denotes the option of no amino add residue being present at that consensus position, and the light chair; variable region CDR2 sequence comprises'an andnrraeid sequence selected from the consensus sequence: tA.D! A S iSFQ ID NO; Ϊ Uo. ht another embodiment. the heavy chum variable region CDRi sequence comprises an amino acid sequence sdenied from the consensus sequence; 0 F O’,St (FJ..4 ;S.N; {j.S.H) i¥,H) (SFQ ID NO: i 17); and the light chant variable region COR! sequence comprises an amino acid sequence selected from die consensus sequence; Q i D.G.St (ΙΛ’) DO) iRO) »A.W:Y> (SFQ ID NO: I IS). in another embodiment, isolated antibodies of the invention bind to human (5D27 and include a heavy chain \enable region including an amino arid sequence selected front the group consisting of MHO ID \’Qv. <\ ", .)-1, 25, Go 57, 4H. v-g qq, ol, "2, AG G-o hr 102. Huh and co«s<· native sequence modification- thcteoi ('be arm buds mas hue her include a light chatn sarutble region including -in armno .tv. id sequence selected I rum the group eon-t-bng of SFQ ID SOv 12. id A. 19. 50. <1,42. -I h 54. 55. w·. GF 78, Olq OF % 07,. I OS. tO'h .aid conservative sequence m'-dirieanons theteuf. in a stid lurtbor emb-.'Jinicnr tv.dated annhodte.s - 4' die mx-cubon bind to human OD27 and mrlude a heavy chain anubk region and a right eh.no v.triable region including the. amino. acid sequences selected from the gr<mp cons!sung of: tat SFQ ΤΠ NO: 57 aud/ur 45. mspeetr-ely. and censers stive sequence mod i iloariotis thereof:,

Cb) SEQ ID- WQa; 49 and/or 15, respectively, and conservative sequence :n:50diiieatk>ns thereof: ίο; SEQ ID N()c Mj.) and/or HR respectively, and conservative sequence modiflcari on.s I hereof: (d;f SEQ ID NOs: 85 and/or 91 and/or 97,5sj$peeq:cely, and: conservative: sequence 0¾>ttitlcatloos thereof: tc s SEQ ID NOs: 25 and/or 31, respectively, and conservative sequetipe modifications thereof: i f> S P.Q 1D HOs; 73 and/or 79, respectively^ and: combrvaiive secpence (g)SHQ ID NOs. 7 and/m I 3 and/or IQ respectively, and conservati ve aeqnsnKv modifications theR'ot; and ins SEQ ID NOs: 6! and/or 07. re^|^cΓίvelyϊ.:¾»d.C·^ιlsφ3ί½'a!.ίtγ·e'.$e¾taeήcd:: modfSeaiions thereof:

Iso sued antibodies v, hied Include heavy and light chxm variable regions having at have 807, or as least 85N\ or at least 0(77, or at least 95q. or <*< least %'v. or at feasr 07(7. or at least 68 A, or at least 9yfA, or more sequence identity to any of I he above sequences are also itsdtided lit the p.esem invention. Ranges intermediate to the above-recited values, eos. heavy and light chain venable regions bas ing ot least 80-8571. 85-0()0, 90--95N or 95 -10(.:7 sequence identity ίο any- of the above sequences are also Intended to be ene run passed by the p-e.'Cnt invention.

Also encompassed by the present inveniion are Isolated antibodies Much -compete for binding to CD27 with the amUxxhes oi the invention. In a panic-dor embodiment, the amthody competes for binding So CD27 with an antibody comprising heavy snd/ur light chain variable regions comprising the ammo acid sequences sc* forth sn SEC} ID NOv. 57 and 43, SEQ ID NO*: 46 and 88, SEQ if) MV 103 and I ID. 5EQ ID NOs: 85 and 61. SEQ ID NOs; 85 and »>?, SO} ID NOs: Ε> and 51. SEQ ID MV. 73 and 79, SEQ ID MV 7 and 15. SEC} ID NOs: 7 and 19, 81:5} ID NOs' (·»! an 67. respectively. or amino acid sequences at least 80A identical thereto in another emRxIimetn, the antibody competes for binding to CD.?. 7 with an antibody comprising heavy and/or light eluin variable regions comprising the ammo acid sequences set forth in SEQ ID MV 57 and 43 (1F5), SEQ ID NOs: -Id and 55 11HS) or SEC} ID NO-: 103 and 106 (5H12). In another embodiment, the antibody competes for binding to Γ027 v, nit an antibody composing heavy and/or light chain variable regions comprising the arnino acid sequences set forth in SEQ ID NOs; 25 and 31 OC5). SEQ ID NOs; 7 and 15 -5HSE SEQ ID NOs: 7 and E? OH8; SEQ ID MV 01 an 67 t NO) or SEQ ID NOs: 7> and 79 (206). In ye· anothe· embodiment, the antibody composes for binding to CD37 with an antibody comprising bear y and/or lights ham variable regions comprising the mm no aend sequences set forth In SEQ ID N'Os: S3 :md 41 ; 8A Ml) or .81:.(} ID Nf)s: 85 xml 67 t 3A ! On

Orbcr antibodies of the Invention bind to an eplropc on CD27 recognised by The antibodies described herein. In another particular embodiment. the antibody binds to an epitope on ΟΠ.?7 rec-gni/ed by an antibody comprising hears and/or ban; chain ;-triable regions composing the ammo add 'Vqnem os set forth m Si Π ID '\t>o s~ and 45. SEQ ID NOs: -19 a?d 55, SfQ ID '<0< 1 05 and 199. Sj-Q ID NOs; 55 and {M, SEQ ID NOs: 55 and 97, Si:':0 ID MX: 25 and 31 SEQ ID NO: -5 and 7'K SEQ ID NO: 7 and 13, SF.Q 0) NOs; 7 and i9, SEQ ID NO\; (d an O'". re-pccfoely or ammo acid sequences at East 80D· identical the; cm. in a; to the; emlvd tenon L the antim-dy hinds to an op nope on CiX:": recognized by an antibody comprising howm and/or ligiu chum sanaHe regains cmrtptismg the amino at. id sequences set lot tit in SEQ ID NO4': 3‘7 ,md 45 i 1 ID), SEQ ID NOs; 49 and 57 ΐ IHH) or SEQ ID NOs: 103 and H,;-b p/Hi./s in .mmhor embwlhnrnt. the anti bod) binds to an cpiiope on (7D3 7 recognized by an antibody comprising, beat ) and/or light i.ham variable regions comprising the amino acid sequences set form in SHQ ID NOs: 25 and 5:1 (5(55), SEQ ID NOs; 7 and 1 5 (3H3p SEQ ID NOs: 7 and 19 <3HSk SEQ ID NOs: 61 an 67 i .1G5) or SEQ ID NOs; 7.) and 79 t5G9), In yet another embodiment, the antibody binds to an epitope on ("037 recognized by an antibody comprising heavy and/or light chain sartahie regions comprising the amino acid sequencer set forth in SEQ ID NOs; 85 and 91 :3A10; or SEQ ID NOs: $5 and 9? OAiO),

The .antibodies of the ftiveatiotr dan either be itnl lidepgihx 'lbr example, any of the following tsoiypes; tgGl. \g02. \gCXX IgG-l IgM IgA;. IgA.?.. IgAsec, OD. and IgE. Alternatively, the antibodies can be fragments such as an amlgen-bindtns portion or a single chain antibody re.#,, a Fab, Rabh?. Fv. a single chain Fv fragment, an Isolated complements*dy determining region iCDR) or a combination of two or mere isolated CDRAh 1'he antibodies can be any i.md ot antibody, iutiitding, hut not!united to, human, human-zed, and chimeric anti bodies,

Tumor antigen*·, employed by the present uncut Ion b\e , m a vuc-eme, used in combination with an anti-CTED antibody of the mve-ruton i include ,uty antigen o; anngeuie determinant which n prc'cnt on tor associated %\tth) a tumor ceil and not typically on normal ceils, s>r an antigen or anhgen-c dote:mutant which ts present o« or as-'oeiated with inn re* celts h: greater ant*'unis than on normal t nmt muuort tells, or an antigen ot ammonia dcsenthnatu which is present cm tumot cells ut adUYeicnt iorrn than that lonnd on normal !not;-tutriot'i cchs. bitch antigens include tumor-specific .rumens including tumor-specific membrane antigens, tumor-n-soemted antigens, including tunics - associated membrane atuigens, etni.'t yotuc unbgorss on tumor·:, growth factor receptors, growth I act or ligands and any ether type of antigea that is associated: with cancer. A tumor antigen may be, for: <.'\amp!e. ,U5 epithelial raneei snuge-i. !C.g.. breast. mofTOmtesunai, fnngs.. a pm.-state specific wincer anttgen i.PS.A) or prosaic «per 1 Vc membrane antigen fPS-MAr a bladde- ounces antigen, a lung \r.<!„ umht cell lung- canees antigen, a colon cancer antigen. an οι cruet canee- am I guru a brain cam er arm gen a panic cancer antigen a none cell earcmmtia anttaen. a pancreabc cancer antigen. a U\er cancer antigen. an esophageal cancer KtUigom a head and neck ounces antigen, or a eokeecub cancer antigen. For osatnple. the antigen may. include ,· tumor unt-gcti such as pisCii. go 100 <>f piste! 17, CF.A. eg 100, f RP 2. NY HR-1. NY -CO-58, MN tgpdbO's idlotype. Tyrosinase, Tekrmmse, SSX2. M1JC-!. MAGE-A3, and high molecular weight-melammua associated antigen iHMW-MAA) MaRTI, me)an-A, IsGERviih NY-ESO-L MAGE··!, MAGE-3, WT!., Herb,or -nesothenn. Other antigens employed hy the present invention {»>,£.. in a vaccine. used in combination xvith an anfi-CD27 antibody of the insentient! include unfigstns from infection-- disease pathogens, -arch as viruses. bacteria, pamsifes and fungi, examples of which are disclosed herein.

The invention also provides a hi specific molecule comprising an uittib-.tdy <>r the invention linked to a second in notion;;.! moiety having, a different binding spec; licit v than said antibody, For exampie, in one embodiment, the second molecule may bind a T sell receptor imp. CD3. CD41.n.

Composition;· inch.tdittg an anribodyor a hi specific molecule described herein, lorn at I mod with a pharmaceutically acceptable carrier, u.tv also provided. Thu compositions mav iunher include an adjuvant, irnmuno.srimuiaioj'y agent (e.g., CP40 ligand, |::LT * ligand, cytokines, colony-stimulating factors, an ami-CTLA-4. antibody. anti-PD! antibody, amt-41BB antibody, ami OX-TQ antibody. I,PS mndotoxin·. ssRNA, dsRNA. Bacilic Calmette-Guerin tlTCG). Lev a ml sole hydrochkn ide. intravenous irnntnne globulins and a Totl-like Receptor (TLE) agonist TLR> agonist such Poly 1C. a T1.R4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist. and a TLR *·> agonist)), smmunosuppressn.e agent, another antibody, or an antigen. Exemplary antigens include, but are not limited to, a component ;)f a pathogen, a tumor antigen fry., lihCG, gp 100 or Pmc; I T", BElCVtieu. WTL mesotheiin.CEA gptOO, MARTI. TRP-2. melan-A. NY-ESC-1, NY-BR- L NY-CO-oR, .MN tapirm·). idlotype. MAGE-1, MAGE-3, MAGE-A3. Tyrosinase. Teiomemse SSX2 antigens, MGC·· I antigens, and germ ceil derived tumor antigens !, an infectious disease antigen (e.g. viral soil gens, bacterial and parasitic antigens t an allergen, or an a.utoamigen, .Assy of the antigens -disclosed herein can be included in a composition of -he invention.

Nucleic acid molecules encoding the. amlfeodfetiof the towRCiors are also encompassed m the hwendmt, -w well us expression vectors comprising such nucleic acids, and. ho'i colls c-nmwNmg syeh cxpsesxiors -vectors. For example, m one embodiment the nr·, enttot; presides rut isolated monoclonal antibody that binds human (1.)27. wherein the antibody comprise··· a heavy chain \ ariaNe region and a light chain van aide re mem encode:.; by nucleic acid wwinencex selected iroro the group comma my of; pit SF.Q 10 NCK: 5 and 1 1, respectively; (b t Sb.Q ID NO·: 5 and 17. wspec to eh: ml S12Q ΙΠ Mi is; 27 and 21, respective!y. td) SDQ ID NOs; 77 and 41, respectively; te) Shi) ID N'Om 47 and 77. respect reeh; if) SHQ ID NOs; SI and Pi. respectively; tg; Sf,Q ID NO: 7 l and 77, mspcetivdy; ilt; SHQ ID NO.; 1¾.¾ and hi, ( = > SDQ ID NOs; 87 and 15; φ SBQ ID NO*' 104 and 117.. respectively or nucleic m ;d sequences having at least HO identity to the nucleic aei-J -equence·- of taotbt.

In another embodiment, the pie-sent invention provides methods tVm inducing Of enhancing an immune response ;<..ip. a T ode mediated nmmme response. and/or cm NK-mediated espouse anci/or a 8 edl ntediateU immune response} aealnm j.r; .uni.yen in u subject Pv admin istenny to the subject an rrYecme amount of an mm body u,.e,. a Util length antibody.!, composition or bispreific ntolecnle Jose; ibed herein. Such methods are particularly well-suited for use in vaccine therapies.

The antibodies and other corn posit tons of the present invention cars also be used to inhibit growth ol'OD27 expressing ceils by contacting the cells \s hit an ant-body or composition in att anion.nt effective to inhibit growth of CD27 expressing cells 'I·,,;’., in the treatment ot cancers?. Anybodies useful in inhibiting the growth of CD27 expressing cells include full length antibodies and fragments thereof, as well as antibodies that contain a second binding specificity for an be receptor. 1st cate embodiment toe CD.2" expressing cells are contacted with an antibody in the presence or'effec tor cells under conditions sufficient to nidnee anhbody-depsntdetit cellular cytoxicity ι ADCO of target cells (Y.y.. the ami I tody induces at least about 40N specific lysis of CD27 expressing cells at a concern! niton of lOuy/ml and comprises Si/.Q ID NOs;6l, 67, 85. 91,17, 37. and/or 4 N. Is; another embodiment, die cells are contacted with an antibody under conditions sufficient to induce, complement mediated cytotoxicity (CDC) of the ceils ury., the antibody induces at least, about 40¾1 complement mediated cytotoxicity (CDC? ofCD7.7 ex pre-sing ceils at a concern ration of )0 ug/ml and comprises SEQ ID NOs; 7. 13, 19, -|1„ 55, 103, and/or 109), embodiment, the antibody employed, lo inhibit growth at CDS? expressing ceil·.-; mas also possess (or lack ! additional Itwuional I'eauues. For example, the antihods may also inhibit -he binding of C l'5?!] to CD?7 on cells which ex press these proteins i<,g,. a mAh compriswg hoax y and/or light eh, an \ ariahle regions sequence^ cwo posing SEQ ID N’Ov. 37 and/or 4/ < m Ao IΓ5 8Γ.Ο ID AOs: -id and/or ;o (mAb IBBi.orSBQ ID A(V; 103 nnd/x't 100 unAh 5H 10·. Alternatively, the nnobudy may not white the bind mg oKΉ70 to CD.?? on such ceil-, u-.e . a mAh cmuprislng hew) and/or light cham variable regions sequences compos mg SEQ ID ΑΠν: H and/or 67 tmAb lG5x SEQ if) AOs: 85 and/or vl. 85 and/or A? tm.Yo 3Λ103, or SEQ ID AOs;? and/or I 5. 7 and/or 16 tmAb 58181. CD?? expressing cell·' include any and all cells the express CD..· 7. including, hut not limited to NR cells. B cells and T cells, in a particular embodiment, the CD27 expressing cells Indude cell lines such ns iiui.ai ceils, Raji cells, Ramos cells ;utd Duudi cells In atst-ther embodiment. the CD27 expressing cells are tumor eebs or cancer edb.. irs another embodiment. CD?? expressing usIK indude B cells, NK cells, Γ cells that are found 'wiilrrattng tumor or cancer ceils, also Called tumor mlilfratmg lymph··.a.vies.

The methods oh Inhibiting the gross th of (TD~ expwsxmg cells that are described herein can he' used to treat and present a Ά ;de variety of diseases and disorder?.

Dor example. In one embodiment the methods can bo used to new or present a canoe- u op. a Cancer selected hiom the pomp c-mststing of leukemia, acute' lymphocytic leukemia, acute myelocytic leukemia. mvdohiaxA promyelocyte myelomonoeytii monocytic erythr'.deulemfa, chronic leukemia chronic myelocytic tgramdoeytic! leukemia,'ctenid hmphoevhe leukemia, mantle celi lymphoma, primary central -teivous system lymphoma, Burkin's lymphoma. marginal zone 8 ceil lymphoma. Polycythemia vera Lymphoma, liodgl in's JiS'-asr. non-Hodgkin’s disease, multiple myeloma. Waldenstrom's mncioglohulmemia. heasy chain disease, so!id tumors, sarcomas, and carcinomas fibrosarcoma. myxosarcoma, liposa.reoma, citsondroseucoma, osteogenic sarcoma, osieosaicmna. chordoma, angiosarcoma, endotheliosarcoma, lymph,ingiosarcoma. IxTUphangioendotheliosareomu. synovioma. mesothelioma, LwlngX tumor, ieiuroyosamomu, rhabdomyosarcoma. colon smeoma, colorectal carcinoma. pancrealtc cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, xxxent gland carcinoma, sebaceous gland cavernoma, papillary c,t? vinosfta, papillary adenocarcinomas, evstadenoearunocna. medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duet carcinoma, choriocarcinoma. seminoma, embryonal carcinoma. Wllrn's tumor, ct-rvical cancer, uterine cancer, teshedtar tumor, lung carcinoma, .small cell lung cam!noma, non small coll lung eatchvotna. bladder cna'inuma. epithelial carcinoma, glioma, astrocytoma, mediilloolasmma, craniopharyngioma, ependymoma. piticaioma, hemangioblastoma. acoustic neuroma, ol 1 godendroglioma,. pienangioma. melanoma. ommoblroomo. retinoblastoma. nasopharyngeal carcinoma, esophageal carcinoma, basal cell carcinoma, biliary tract cancer, bladder cancer, hone cancer, brain raid central nervous system. ;ONS t cancer, cervical cancer, chinnocaresnoma, colorectal cancel s. connective tissue cancer, cancer of the digestne system, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer. Intraepithelial neoplasm, kidney cancer, hrsm cancer, liver cancer, lung cancer (small coil, large colli, melanoma,: neuroblastoma: oral canny cancertlor example kp, longue. mouth and pharynx!, ovarian < ancer. pancmnic o oncer. miinobkntotoa, rhabdomyosa-eomn, tecta! c.utC'.-r; cancer of the : expo more -g,\rem, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine emu or. and emu or ot die urinary system) Preierred cancers iuclude 0 CO 7 expressing. ttnnors selected frotn the group consisting of chrome lymphocytic leukemia, mantle cell lymphoma, primary centra! trr.vus system H muhoma. BurktU's lymphoma and marginal /one B celt lymphoma. In another embodiment, the method'- can ho umd to treat or present a hauerial. limgal, xiral or parasitic ίαίοοτΚ-η.

Tlu- present invention further pi os ides method··· for inhibiting the binding of CD7U to f Pd? on ceils its a subject having a disorder by administering m the subject u ott bodies or coni positions a*· do- ended herein, ns oed us methods tin drmn-tvgukttiug a T cell response in an individual having a disorder by administering to a subtect antibodies or compositions described Herein. These methods are ideally suited tor use in the treat mom of immune disorders, such as graft ιejection, autoimmune diseases, and allergy. Antibodies useful in these methods include Fab fragments. as vvcll as a mutated fc region so pen me antibody does not bind, or has significantly reduced binding to. be iccepiors . In a particular embodiment, the antibody comprises heavy and/or light chain variable region-; sequences comprising SEQ ID NOs; 37 and/or 43 (mAh 115r SEQ ID NOs: 49 and/or 53 tmAb DIBi, or SEQ Π ) NOs; ; ()3 aud/m 109 tm/Xb ?H 12b

The methods described herein tor inhibiting die binding of CD70 to CLO? on cells and far dosru-roguiaime a T cell ;espouse can be used to treat a '-vice variety of diseases and disorders, including, but not limited to graft rejection, allergy and sunnmnume diseases. In a particular embodiment, the disease is an autoimmune disease* m.,e., multiple scleiosis, rheumatoid arthritis, type ! diabetes, psonaHia, Crohn's disease and other mtlawtmatery bowel diseases <-ueh as alec-rath e colitis, systemic lupus eyihem&amp;tOKus iSUia autoimmune-encephuUinm'hti··, mv-jusihenia gravis (MO), HashimctoS. thyroiditis, Goodpasture's syndrome, pemphigus, Ciraves disease, autoimmune hemolytic anemia, autoimmune-thrombocytopenk purpura, sdeo.-denna with ami - collagen ami bodies, mixed connective tissue disease, polyp} ositis, pernicious anemia. idiopathic Addkon’v disease. autoimmune associated infertility. domerulonephniK crescentic glo:ro:rukmepie;U\, pioliferattve gkauerulonephritis. bullous pemphigoid. Sjogienk syndrome, psoriatic urthnttx insulin tfststemee, autoimmune diabetes rnelhuo, muohninune hepatitis autoimmune hemophilia, anti-itututme lymikiofmdlfesaase .-yndrotne {ALPS», ainomununo hepatitis. jutuimmune hemophilia, auioimniunc iympbuproiitvrative syndrome autoimmune uveoretinhis. Gu'dklbt-Bure syndrome, aneriosderosi-- and Allien tier's diseased.

The present invention further provides tor particular uses lor the antibodies, compositions and hi specific molecule- described herein, bor example, m one embodiment, ihe i;r- enikat pros ides for the use of mi anti hud} composition or bispeed'ie molecule In the rtianuiactweidf'a medicament for mduclng or mth&amp;iwfng an immune response -agiaiitsf an antigen in asphjfiph ln::iiigh^r-.e{ln|K>4l.}ipppMi the Invention provides for the use of an antibody nr composition m the manufacture of a medicament for inhuming growth of ΠΧ'? expressing cells, the use of an antibody or composition in the rmmnUeiure of a medicament for inhibititnj the Lending of €'D?0 to CD27 on celk nt n subject having a disorder. and the use of an ailtibodv or composition in the otattulacturv ol a medicament t’oi down-rogtda?mg a Ϊ cell response in an individual basing α disorder. The presec? invention further is tel ticks an antibody, composition w bispecific molecule for use in inducing m-enhancing an immune response against an antigen in a subject, an antibody or composition for use m inhibiting growth of CD27 u\pressing cells, an antibody or competition for use in inhibiting the binding of CD7t.) to CDc / on cells In a subject having a disomer, and an antibody or composition for use in down-regulating a 1 cell iespouse in an individual having a. do-oclcr.

The present Invention also p/ovidcs methods for detecting the presence or absence of CD27 in a biological sample by 11; contacting a biological sample with an antibody described herein in herent die antibody i-· labeled with a detectable substance) and \2i detecting the antibody bound to (11)27.

Also within die scope ot the invention are kits comprising the compositions ury., antibodioN and/or hi .specific molecules; of the invention end. optionally, instructions fur «se, Πis: kit can further contain a least one additional reagent, such as a cytolrtnedr complement. or one or .more additional antibodies ot the invention.

Other features and advantages o(' the instant invention will be apparent from the follow mg detailed de,st notion and claims»

Figure 1. depict-, the afftmiy andLmotk parameters for mAh- KV\ 11½ 3Π12, 3H8. 20b', sFA 3AAh ?C2 ni\ I \~k ms 9F1 and ms ΜΊΑ7i as deremvtned by

BtaedfeyM BiaEvhltsadsn software (Biaeore 'Q>27 ImmoMilixed: ms the chip

Figure 2 is a graph Asms mg the binding of human anti-CD.?.? antibodies (2C2, 3'iiB, I F,\ IG5. i HA 20 A 3A I p and 51! I ? > to fecmmmxam puntied human CP.27 using. MUSA.

Mgoro3 is a graph shoeing husdmg by BLISA of IF5 to pitrifiedm-ombitunn human or monhex· i macaquei CD?7

Figure 4 is a graph depicting the effect of human anti-CD?? antibodies (2C2, 3H8, IF5, 105. .IMS. 2G9. 3AM) and 3H I?) and MslgO's; l A 4. OR- and M-T2? 11 on the binding of soluble CD70 tsCD70) to CD27 protein (shown as A block mg} by ELISA.

Figure S is a floss cytometric analysis of lt;5 binding to human iyiuphobhisn.'id cell lines» und blocking of sCD?0 binding.

Figures 6,4-4) are graphs showing the binding of human am 1-0027 antibodies <2C2. 5HB. and IF5) to CD57 on jurkat evils {Figure 4.4), Rail cells (Figure 4Bs. Ramos cells (Figure 4C), and Daudi rolls if?cure 4D) ns assessed by flow cytometry.

Figure ? ;h a graph showing the funding of human anti-CD?? antibodies <2(52, 3HA .1FS. JG5L 1 MB, 2G9, 3 A10 and 3H i 2) to (51327 on Daudi cell· as assessed by Hove cytometry

Figure 8 is a bur graph showing the results of an aml-CD27 cross-blocking ELISA experiment.. demonstrating that antibodk-s. iFS. 1HS arid 3HI2 are capable of crossblocking each othes and thus bind κ· the same epitope.

Figure 9 is a bar graph showing the results of an ami-CD77 cross-blocking ELISA experiment, demon-a rati up that antibodies 202. IH8; 1G5 ami 2G9 are capable of cross-blocking each othe- sod thus' bind to the same epitope.

Figure 10 is a has graph showing the results of an anti-002? cross-blocking-ELISA experimem, demonstrating that ihe binding of antibody 3A i 0 to CD27 is not fully cross-blocked by any of the other an?i-CD27 antibodies tested, thus Indicating that 3Λ Hj binds a unique epitope, but 3AI0 binding is partially cross-blocked by antibodies i u5. 1HB cmd oHi 2. indicating that ihe epitope for 3AI0 may be clo;-e to ihe epitope bound byanti bodies IF5, IMS and 31:112.

Figure .1.1 is a graph depleting the results of a complement dei.tera.ktm cellular cytotoxicity iCDCO assay using roAhs IK\ 202. 3FI8. lC1f>. 1 H&amp;. 2Cr>. 3A10 and 31 i 12.

Figure 12 ts a graph depleting the results o6-glMit|fer^0^l'emkJ|:##:hdep:t. cellbkr.b^btoxicuy (CDCC) assay using mAh IFf

Figure !3 is a graph depictiny the results of an antibody dependent cell-mediated cytotoxicity tADCC) a«say using mAbs 2C2. IP5. olid. iGm I.H8. 209, 3A10. 3HI2. Ritusan and HuigG.

Figure 14 is a graph depicting the results of a further antibody dependent cell-mediated cytotoxic ay lADCC? using mAh IF$

Ft ««re 15 n an alignment of the \Ή sequences of hitman ami-0027 antibodies {IFF KG, ms. 2?'2. 201 3ΛΙ0. 5H12 and 3H8

Figure 16 iv an alignment of the YL sequences of human aotFCI02;7 antibodies i i F5. IGA HIS 2122, 2G9, 3A 10s 3HI2and 3.H8s.

Figures 17 him! 18 show results l rein an in vivo non-human primate study using. mAh HF8 In purUcular, Figure 17 shows 1 F5 on circulating lymphocytes after a Single dose. Figure IS shows 1 ft does not significantly deplete circulating lymphocytes. 'Figured9 depicts the sesutts of a pemamer st.iimnc assay on mouse periphe; rd blood cells and splenoeytev

Figure 20 depict'' she re''i.Ots:'dffab;EMSFOT:^ssdy: and enhanced IFNy production using ami-CD.;? antibodies.

Figures 21 shows enhancement by aiui-C027 m.AN oi I cell responses »o a yacehie. sofigep in a traiisgenlc mouscmodel by peiviamer staining: atidlFM ELJSPtfr.

Figures 22A-C a-c die protocol for and results or an experiment showing dun anh-CD27 enhances T red responses to an A PC-targeted vaccine p.ol)!2f72b5 -OVA ?, Figure 3.7.A shows the protocol for the experiment. f igure 2.2B shows toe results of a tetrasese? staimoc experiment ?u measure antigen sped He T cells. Figure 22C shows the results of an 1FN-gamma OJSPOT assay to measure ramgen-'-pec-iic T cells.

Figures 23A-D are site results of an experiment showing that anti-CP27 it? combination with the TLR3 agonist PolvlC* (at 22 μη. 50 ng or i00 μβ> enhances T ceil responses to an APC-ntreeled vaccine i(Y.-DF.C20>OYA';, 11cure 23 Λ is a ptapit showing the 92 of 1FN gamma positive ceils among (2.0be- T cobs tor cither wild-·>pc otice treated with poly TC and the ami-CT>37 mAh 1F5. buCD27 -transgenic mice treated with poly 10 and a control human IgGI antibody or huCD27-transgenic since resed with poh 10 and die .oui CD2 7 mAh iff

Figures 24 and 25 show resell·-· from a cuds of administration ai am GD27 rnAb prior to vaccine in the presence or absence of T'.LR aanosi and show the significance ot timing of the. aminis traiio n of the an d body relad v e. to the vaccine.

Figures 26 and 27 «how re subs iron! administration of ami-CD27 m.\h in combination wish TOR activation ion. T-cdis from burr;ns; 0027 transgenic mice, ;o shown by hoth prolifemtion curd mm ok me production

Figures 2H\~0 oc mo piv?· moi for end result'' or on experiment showmg doe anii-CD2" enhances th> --001 a-DF.C20?-OV,\ vuomne in u VI04 ι Η ίο OVA1 meiunmiia d ud tenge rnobel I moo 2 1A -howw -he pfotoeol m-r ihe oriT-'iin-rm I nmn 248 ό a etaph pkiif.no' the mutot w/e Os; mm'1 nan mm rmmbei'ot'davs pom mmoi hwcmaikn >n untreated mice, name 240 is a graph ploifme dm tumor sire 'in umu‘ i against nomom m d;.o,\ pom. turner tnoeubtion in mice υrated w oh the 1.00mo alone. I An re dm-0 m a graph pt-onng the urnet >me pm omd's omen-I number -..-f daw p"si mine» utoenlattoo in mice irs-awd 0 oh me vaccine In eombiuvion 0 U'n an ami-CkO / andhods

Figures 26Λ ami 8 m&amp; gra|iha showing prolonged survival os human €027« franagenk -mice -(tumor models) Mlowtbg challenge with a ayugeoele lyikpltesiia sad administration or van mu, doses oiaml-CDA/ nt-\b i F5. figures 30 to 32 show Ihe results of an experiment resting the effect of amh€D27 treatment in a Rap xenograft model in SOD mice, figure 30-3 plois tin- tumor m/u dn mm ί again·-? number ol -Jays pos· forme; mocuimmu in rtuce either unhealed. heated with a eoturol human iaGl anttbody or treated, with the iutfi-Cl C' U'pand 3H8 antibodies. Arrows indicate day-, when antibody twmmem was given by ί.ρ. injection.

Figure308 shows survival in a Kaplan-Mese· plot. figure 3 i A plots ihe turnur si/e (in mm '1 against number of days post tumor ihoeulation in mice either anueaied, treated with a control human IgCl a nobody or freakd with the ami OD27 If 5 antibody. Arrows mdieat* days when antibody by p injection Figure 31B shews survival in u Kaplan Muter plot.

Figure 32 shows the «suits of a fui ther experiment testing the eftect of ami-CDS? ueatmem m a Rajs xenograft sumteTm SCTD mice hi a Kaplau-Mete; plot.

Figisre 33 .shows the results of an experiment testing ihe effect of ami-CD?7 treatment in a Da ad; xenograft mode! in SOD mice. Figure 33A plots the rumor sure nn ntnt ; against nunthes of days post tumor inoculation in mice either treated with a control Ptao an tgfj | antibody or treated with the anti-CI.)-? ] FS anti bod} (0,1 rug or 0.3 mg).

Arrows indicate davs vt [ten antibody treatment w as given by i.p. injection. Figure 33B shows survival in a Kapian-Meter plot.

Figure 34 --how s the results of an FLISPOT assay and that enhanced antigen·· .^peeMc'll'Ns production usuis anti-CD?7 antibody is abrogated when the Fe portion of the-IgG is enable to engage Fc receptors.

The ptesent invent tot; provides ami-CDS? antibodies that exhibit particular functional properties correlating with significant therapeutic benefits, including upregtdation of immune function te.g. T' ceil mediated tmmane responses as in vaccine therapies. NK activation in cancel therapies), inhibition of cell growth Dye., tn cancer therapy) and down" reanhuion of I cell mediated imntune response;; uyy. in autoimmune therapyg These functional features include. for example; {I s inhibition of ie.g . completely or partially blocks) binding of soluble CD7Q to CD.?.? expressing cells bs at least about 70¾. further for example by at least SOD oral least TOD, {?; hi tiding to human CD? 7 wuh a KD of I x I (Τ' ?vf or less, (3) induction of at least about 4i/-f complement mediated cytotoxicity tODO of CD77 erspressittg cells at a concentration of SO pg/rnL (4) indnetion of at least about difo specific lysis of CD77 expressing ceib; by A DOC at a concern ration; of )0 pg/ml (further for example at least about 50¾. at least about 61)¾ r.u at least about 70¾ specific lysis-(5)

Ittdnotion or enhancement o! trurmme responses, especially ΤΗ I responses and/or (6( induction or enhancement ofT-cel! activity, especial)} specific CObf T -celt numbers and/or activity. In othet embodiments, the aobbodtes include particular heasy and light chain variable regions and/or CDR sequences.

In otdbr thai the f resept invenifon may be more readily -understood* eettafo Mnm are first:defined-. Additional defiiititbns am set forth througlhout the detatled description.

The tents "(ITT.'?'" taiso serened u:> as "Ci.)27 molecule", "CD2”L receptor'. "SI521". "Tcell activation asmgers {.'[>77" "TNFkSF?," "MGC20 FFF" "tumor siecrosls tuctor receptor NupertasrsstY. member 7". "T cell activation andgen Si52" 'Tp55'', "Tumor necrosis !acis.>r receptor mped'amity member 7'. "CD 27 anti sen \ a set 'T-cril activation antigen CD27") mfors to n receptor shat Κ a snentl'Of of the TNT'-receptor supcrian'tiiy, winch hinds ns ligand ΓΠ?0. (!(>27 ρ, reomt'ed fen' gersvrusion and king-seem mans set ns nee -of T ceil hnsviontty are] plays a Lev role 1st regulating B-cell ne'nm.nn.ut end Imsmmoglormlm ay mhos; o The tern;: 't'D27'" oh: lades mss variants os' koforrus of CD27 ehe:it are nn-oraky eapremed by cells se.gw fauna a CD27 iienosbed with GtiNBAAk:P having accession no. ΛΛΗ12 IbO. I >. Aecmd'usgly cmtiFodiea or the invention may cross-reus,t w n.h CD2? i'rosn species miter than htmtan Alternatively, the antibodies ntay be «pecilie lor loasum CD7.7 and may me exhibit any t rotooracsk its it ls.h tuber species, Cl)27 or any variants and isoforms thereof mas elthet be kolated from cells re ;names which naturally cypress deem or he iveombmantly produced using well-known techniques m the ast .anchor those described heresn. Preierably the antibodies are targeted to kCD2? which has a norms! giyeuaylnt-on pattern.

GenbaakG - Accession No. AAHI2tbO.it reports the ammo acid sequence of hnmao:CD27 as follows {SEQ ID NO:! >: II stsiatpb pwwle klgtlvg! s a. tpapfeseper hywaqgMce q.mce pgtflv Ikdcdqlht'k: aa hi qcdpetpgvs fspdhhtrph cess.sin.nag Sk media naeeacmgw qordkeetee 12.1 dplpnp.-lta m-agdsphp qpthlpyvse nslea· taghm cntadf-qlp ardsthwpp '181 qr-.lscsdl's oh'dagm? ivkkigaii Ihiprkyrsii bgespvepae peryaepree 241: egan p mod yrkpepsttesp

The ienn “CD?!)" iako re i erred lo as "01)70 sncdcni.de", "CD27L”, "CP27LG'2 'TNPSF7,'' "tasnos raterocm tactos' (ligand; super fa roup member ?." '721.72'? ligand,"' '"CD70 antigen," "surface antigen {.'[>70." ''tumor aests'oms Sac lor ligand stmer family, member 7," v'Ki-24 antigen.'' and "CD27-I7'i refers to the haand for CD?.? {sec, tor example. Bowman MR rds/..,/, hniMtioi. H/M Feb I re I xitG: 1750-61). CD70 is a type (1 imapnembrane protein that belongs w the tumor necrosis factor t'TNH ligand family. It is a surface a ms gen on activated T and B lymphocytes that induces proliferation of co-stimulated I ceils, enhances the generation of cytolytic 7 cells and contributes to T cdt activation. It has also been suggested that CD70 plays a role in regulating B-ceii activation. cytotoxic function of natural klliei ceils ansi immunoglobulin synthesis fHIntrers RQ «·/ o:L, ./. fnrnmmL 1004 Feb :1^2(4):1762-7^.

GenbankCiV t Accession No. NPJ}01243 ; reposts the amino acid sequence of human CD?0 or follows iSHQ ID NO: 2;: I mpeegsgesv rn pygevtr aaivpivagi yiclvveiqr faqaqqqlpl eslgwdvaei 61 qlnhtgpqqd priyvrqggpa IgmOhgpe kikgqlrihr dmyoavhiqv tlaicsstta 121 srhhptikts giespascri siirlsibqg etia'-.p-ltp lirgdtictivitgtiprit tfU tdeiiTgvqw vrp

Tim term 'antibody'' as referred to herein arc I tides whole antibodies and any antigens binding fragment (>.<,, ’ antie.cn binding portion's or single chain the mi T. An “antibody5' refers. In one preferred embodiment, to a glycopiotcm comprising at least two heavy tH; chains and two light (L) chains inter connected by diva Hide bonds, or an antigen binding portion thereof. Each heavy chain is comprised of a hear v chant variable region (abbreviated herein αχ V; = ? and a heavy chain constant region. 1 he heavy chair; constant region Is comprised of three domains, CH 1, CHand CH v Each light chain ;s comprised of a light chain variable region (abbreviated herein as V(p and a light chairs constant region. The lighr chain constant region is comprised of one domain, CL. The V;; and V;. regions cun be farmer subdivided into .regions of hypervariafcility, named complementarity determining reason.- sCDRInterspersed with regions that are ’more conserved, termed frameveurk region;; PR), Each Va and V; is composed of I Dec CDRs and tons· FRs, arranged frosn amino-ietTsttmss to carbusy-termsmss In the following order; l·RI, CDR I, f'R2, CDR2, FK.L CDR E ER4, The v an aide regions of the heavy and light chains contain a binding domain shut internets with an antigen. The constant regions of the antibodies may median; the binding of the immunoglobulin to host tissues or factors. Including sari tins cells of the immune system (eap. effector cells;· and die Cimr component (Ciq) of she classical complement system.

The term "antigen-binding portion" of an antibody tor simply ''antibody portioif')- as used herein, mi cm to mm or mom fragments of an antibody that retain the ability to specifically bind to an antigen m-.ρ., human CD27p Such "fragments" arc, for example between .shorn 8 and about 1500 amino acids m length, suitably between about 8 and about 7-b' mrhm.· acid.:- ru length. snnabh about S to about 300. for msample abm.n S ίο about 2«>0 amine acids, uv about K) to about *0 ,-.r )np a-tuiio acids ;n length. It has been - boo n that the ant teen bind eta function of an antibody eon be perfoemeS hy ho a; cents of a mil-length antibody. Exam pies nt binding bog. memo '·· mean passed vs ithht the term "antigen -binding portion" of an antibody include <0 a ban bagmens, a monovalent i ragmen; consisting of the V;. \ CL and C'H I doom me Si; a Ft ah1)' Segment. a bivalem tragmetu comprising two

Fab fra anient- I inked hy a Sisal ode End ye at bte btngc region: uiS a Ed fragment consisting of die V;: and Ο Ι l domains; tv s a F\ b'.agmertt consisting of she Vb and V;5 domain-; of a single arm uf an anShod), (v; a si.Nb fraginent iWard ua/b (lOb'S; Summ .mj;S44->-iS} which consists of a Vo, domain; ami; vi i an isolated complementarity determining region fCDK s or (vn'i a combination of re. o or mom h-otated ( T)Rs w hich may optionally bo jomed by n synthetic linker Furthermore, although the nto domains ; ?’ the FA tmgmeui. Vb and \γ·. are coded tor In separate genes, thus can be joined, using recombinant methods, by a synthetic linker that enable-.· ibem to he made as a single protein chain in with h the V{ and Y;( mg ions pair to boon monos a!cm molecules {known as single chain 1A isch s; -<ce c.y,.

End et oh > S fig 8 i AV.mm v 2-47:47 V 42m and Huston c? ai. { KCC) /V.··· <. 'Ve/e . bob. Sit. tf 5. I 85.58'γό ή Abb;. Such single chain and bodies am .also intended to be encompassed within the term "antigen binding portion" ot mn anil bods These antibody Fragments a re ohtai nod asiug conseniitni.il ieohniooes known to those with dob in the ana and the 1 ragmen ts ere screened for utility m the same manner as are intact antibodies. Antigen-binding portions eat* be produced by feee;ubio;nn DN-\ techmqaeo or by entymatk, or chcnoo.d cleavage of intact no munoelobulins. A "btspecific'' or 'ImmitOionai antibody" i-. an arSbewi hybrid antibody having two different hear y/hgln chatn pan's and two 'different binding sites FT spec; fie antibodies can be produced by a variety of methods including Fa-amt of hvbndomas or linking of balf fragments. See,.-- e.. Soogsh ilai &amp; Lnchmann, f tin. da/?. lnnnunt>L "We j g -32 | 11SOS g Koslciny e< nJ.. 7, Immunol. 1-48, 1547-1553 10*3.7.5 1 tie. term ''njonoclo-tal ont-nodyf' as uses! he mm, refers to an antibody which displays a single binding specificity end a!Snipe lb; a nanicnia.r epitope. Accordingly, the term "fas mao monodonai antibody" re iocs to an antibody which do. plays a single binding specificity and which has me lathe and optional constant legions derived So an human aermboe immunoglobulin sequences. In one embodiment, human monoclonal autuxxhos aw produced by a hydride;:· a wines: includes a (¾ edi obtaaned bom a frmu, genic non-hunxma aahnub cm.. a traosgen.fi.' mouse. having a yen ο me comuri.dnga human bwn y chain imnsgrne and a light eham uau.sgene fused to an n'omorrahaed cell, I he tenn "recombinant busman meihodyF an u-ed hereon includes all human antibodies that are prepared, expressed creased or donned by recombinant means, such-aila). and bodies isehmed from an a nun a I i ¢-.,4,, a mouse* dm; is Iran'.genic or bomsehronsosouud for human i norm ax sm j shru b a genes or a hybriuoma prepa;eu therefrom. do) antibodies ssnlated f-vu: a host cell u aas formed U) ex pa ess the antibody. e, p.. tVfirn a truoNieetoinu. to antibodies isolated born a mcorrmmam. combinatorial human antibody library, and Ul) antibodies prepared. expressed, creased o- isolated by any other means that involve splicing in human imnmnogicbutm perse sequences to- other DNA sequences. Such recombinant human antibodies comprise variable and constant regions that utilise particular human germ I me immimoakdihm sequences ate encoded by the germ hue genes, Ku include subsequent rearvungesTiems and mutations which occur, for example, during antibody maturation. As km ex a lu the art (see. ep, Lonberg (2005) Numrc Biotech. imp} Μ7Ί 1.7.0-. the variable reason contains the uotsgcn binding domain, which ts encoded hy various genes that rearrange to lorrn an antibody specific fora foreign antigen. In addition to· ivarmngenu'nt, the variable reason can be further modified by multiple single am ism acid changes ; redos ml to as so rant so mutation or hy permutation* to increase she nffnmy of the antibody to the foreign antigen The cm;stem region will change in further response to an antigen , isotype switch).

The tv to re the rearranged and somutienhy mutated nucleic acid molecules that encode the bam cham and heavy chain immunoglobulin polypeptides In response to an antigen may not have sequence identity w oh the original nucleic acid molecules. dm instead will be «ubstantblls iriemiea; or similar rim., have at least SOT- identity's.

The term "human antibody'' includes antibodies having vanabh* and constant regknis (if prevent) o! hnntan nennline immunoglobulin sequences. Human antibodies of the Invention can include amino add .resMues·:not encoded by human sequence' u- g , mntubmix ini;·*deem] by ms-dom or sine -pecuic mutagenesis >:n \ iicu or by somatic nmtaimn in r?wd <.*<·. 1 .onberg, N, a nl t tvs>4s .Voswre .((>S(b474); hhb-dbb0 Leeb'x'sg, "s', i Idd-4 * HmhUx·.-.-( oi Fxpi'nmavoJ Akxn'fmnx'h.-gv 11 3:4V-: 01; Lonberg, N, and Hnsear Ο. :Ibbw) i>u; n< k< r, hni>inii'>L Vol. LL 05 Am and Itmsimg. F ;uul Loaders:, N. * dog s, luu. As}'. Λ; nri. d i ?:'.4;5ef)-e4pi. Bosxm er. the tern- "human anti body” docs not include antibodies In \s Iticn (.'DR sequence·.' tif nv cvl front the get mime of another mammalian .species, such as a πκ>ό§<5, iseetii J^t^ont^-hum^ {!.;:.. hinnsnriicd antibodies'.

As used herein, a "heie-ologous antibody" is defined in relation to the transgenic non-human organism producing such an antibody. This term refers to an antibody having an amino acid sequence or an encoding nucleic acid sequence cone spend my to that tound lit ms organism not eotisisling of the transgenic nondvarnan animal and generally f-om a specie-· other than that of she transgenic non-human animal. .-•Yu "isolated and bods·,'' as used herein, is intended to refer to an antibody v.'hieh to substantially free ot other antibodies basing different antigenic specificities t an isolated antibody that specifically binds to human CD21 is substantially free oi antibodies that specifically hind antigens other ihtsn human CDs?5. Au isolated antibody tied specifically binds to an epitope of may, howevei. base cross-react ο ;ty to other CD27 proteins from different species. However the ant ibody prefer.miy alway s binds to human CDd7. in addition, an isolated antibody Is typically substantially free of other cclliriur material and/or chemicals, in one embodiment oi the invention, a combi nation of "isolate#f antibodies having different CD.77 -.peei fid ties is combined in a well defined composition.

The term "epitope'' or "antigenic determinant'' refers to a. site on an antigen ίο which an immunoglobulin or antibody specti'kaliy binds. Epitopes can be to-rued both from contiguous amino acids or noneomigamw amino acids juxtaposed by tertiary inkling of a protein. Epitopes formed from contiguous aroinu acids are typically retained on evposuie denaturing solvents, whereas enhopes termed by tertiary folding are typically lost on treatment with di-maturing sol-rents. An epitope typically includes at least 3, 4 w p, 7, S.

Hh 11. 13. to, 14 or 15 amino acids In a unique spatial conformation. Methods for determining what epitopes ate bound by a given antibody the,, epitope mapping' are well known hi the art and include, for example, imnuinoblohing and immunopracipi tat I on assays, whereisi overlapping or contiguous peptides trom CD27 are tested for inactivity with the given &amp;nti-CD27 antibody, Methods of determining spatial conformation of epitopes include, techniques in the art and those described herein. I'm example, x-ray crystallography and A· dimensional nuclear magnetic resonance wee. e,g.. Fpiiopc Mapping Ftouktth in Λ/cmw/v m M&amp;tesMMr Vo!, 6 b, CEB. Morris, Ed, flfribl).

Also, encompassed by the present invention are antibodies that bind to an epitope oti CD2? which comprises all or a portion of an epitope recognised by the particular same or an ovedappHig: mgloa cc a region bntwnen or spanning tin' res ton h

Also eneontpassed by the present Invention are antibodies that bind the same epitope and/or aiuibodtce that compete tor binding m human CD.?7 with the antibodies described herein Antibodies find recognize the same epitope or compete for binding can be identified using routine techniques. Such teehnioues include, tor example., on immunoassay, which shovm the ability of one antibody to block the binding oS’ another antibody to u target antigen, Lc,. a compete ice binding assay, Competitive binding is determined in an assay in winch the nmvaiiKy Lmobn under test inhibit s specific binding of a reference com body to a eomsTton anttgen. such as nv". Nmneious types oi e*«mptftf«u binding a‘says am In **u„ ibr example; send phase direct or indnvet rubtoimmunoass y, iRi Ai, soda phase otwa or indirect enzyme unmuuojs'my iΠ \i, sandwich eompehtton assay (see Stahh < / j; , d/eo'c-, A in r - i;A2 i j Ά? i;; --<pd phase dti-ect i'mmnm- nfm I.1A : sc·» Kirkland / m*. e, f.nnHHnui i e" ;af -14 (i Ά·ρ·ν \,ΌΡ otiose dLvef [ weied u,-say, solid phase dtfCrt iabck'ti tumdwicti a-.say o-e-e i iari-.m and ( auc. , Cm A mb ο \ Au/v 0.70,-0 uV.mw/ Cold Sprnn:

Ra-hor Kress ; 1 PS sip -.opd on oedhect i sin > R( \ umig !- i75 Libel t-.ee M--id m <A., e/o/. hnnnuif)1 ? A 1 s ~ ; i! Af i e -mhd pKoe dmwt l>:--ttn astdm i;iA (Cheung w -ji . Vm A, 176:5Tb t Ι^ίΟιι; and Jucm labeled RLA ; Moldenhuud'.-; d A owi j, Ammow/ VI:""' 1 jOSHii! lypiealiy, such an assay or·.ob-e-- the use .'fpnriued am wen Pwtnd t-· a 'olid suti'acc Os >„ His bean my edltet e>i tltesC. an tmlabeled test imo-unosMiuhn aid a labeled reference nnntunociobthtn C empeh-we inhibit;- -o is me mu wo by Jetetmhin;/ the amount of label iituind to the -wiij or face w ceils ns U$c ptcionee of she tv* l unmunoglobiutn l stcdly the tew iiwmwegwfwiw ts present tn esrew. I'suuijy. when a cempebng attttl.'ocis is pm sent n rvem-s, u nil; inhibit specific binding of a reference amibods to a coromoti antigen by at least 50-55--(-, gym-V;-, 60-65A:-, 65-/OR: 70 75A or more.

Other techniques include, (or evmtgbe. epitope mapping methods, such as, \-ray analyses cd'crystals 0; amigemantibod) completes which provides mm the re-wkthon oi the cnitopc Other methods me-mtor Hu* binding of the antibody to anttgen Augmcrsm or mutated \ anutions uf the anttgen where loss oi binding due to a modification 0;’ an utmou a cist iCsidne within the antigen - ewwwe i-' often considered an indication o- an ephe-pe component in addition, computational combinatorial methods fw ep-topc mappms can also be used. Those mcih-.-Js H'iy on the ability of die antibody e-f i mures? to affinity is-date ,'pocific abort peptides itvns c-;n:bin.iionai phage display peptide 1 Amine,-, The pop Odes are bum re^,5rdO'i as leads s'·,·; ristr definition of the epitope Csa70spe>rsdwg to the antibody used to voreeo thv peptide nbrac\. rot epitope mapping, eompuraiionai algo;hhtns base also been box eloped wince have he-";: -.hosx a to map conformational discontinuous. epitope:·.. \s used heooio the terror "specific binding," "selective binding." "sekctix-eU-heads,'' and "specifically binds " mfer ox antibody binding to an epitope on a pmdcrermmed anriyt'ii. T> picady. -he antiboux binds wnh an equilibrium dissociation constant i Κρρ of apptoAiniPieK less than Id M. such as ορρ·χοχ!ο;αη.ον less than 10 Λ Μ, I0’" M or ΙΟ'1"' M or oven lower when demon mud b\ surface piasmon resonance (is PR) technology in a BlACClRR 2000 ms'tmniem usma iceorphinon? ho man CD27 .as ?be analyte and the amibody a.·' the bgand and bind1 to thr predetermined ataigeo v. tth an affinity that: is at least two-fold greater than its at Tin >t> tot binding so a non specs be antigen u- y.. BSA. casein) other than the ptvdctenrunod antigen or a do-els-related antigen. The phrases "an antibody recogm?.;ns an antigen" and "an antibody memhc ior an antigen' axe need interchangeably herein with the term "an ant-body winch binds -.pet.ideally to an antigen."

The terns; ‘IK]»" as: nsed hereitg is inietided to .refer to- the dissociation et;ui hbnem D-iOtunt oi a particular unfibody-nnhaen interaction. Typicalh, the Pun:ass anil Pod ies of the invention Pi-id to ( TX:7 with a dissociation eyxuhbnum constant : hhe of appro'\i;pa:elv 10M sat levs, such as lew than Id M or 10 '" M or even lower when determined by surface pla«nton retsonance iSPR) technology in a RIACORE 2000 instrument using recombinant human CD2? as the analyte and the antibody as the ligand.

The term "kef as used herein, is intended to refer to the off rate constant Tor the dissociation of an antibody bom the anti body Pom gen complex.

The terns "ka" a1-: used hereon is irtfended to refer to the on rate constant for the association of an antibody with the antigen.

The term "HOO," as used hereto, refers to the- concentration of an mshhuds or an antigen-bending portion thereof, which induces a response, eithes In an in rf/o, or an in v;ro a-say which !s 50sf of-he nuomnai response, be., halfway between the max-out! expense and the baseline. A', used herein, "Retype" refers so the antibody class Aye.. IgM or IgOi? that; is encoded by heavy cltatn constant region genes. In one embodiment, a human monoclonal «inibody ol the invention is of the TgGl isutype. In another embodiment, a human monoclonal antibody of the invention is of the igG.2 isotype

Tlte term “binds te ImaioMtldod: COS?," refers la the Ability of a human: antibody of the invention to bmd to Cl V tor exaotpky expmssed on the surface of a cell or which is attached to a solid support.

The lean ~ews· reacted' ms used herein, rotors to the ability of an antibody of the invention to bind to OD.i? front a different species, For example. an antibody of the present invent ton vtbich binds human CD27 may also bind another spades of CD27. .As used herein, oross-reactiylfy is mew-aired by detecting a specific react iv its with purs fed antigen io binding assays iis.tf., SPR. EUSAt or bmdmg to. or otherwise functionally interacting with, ceils physiological I v expressing CD27. Methods for determinism cross-react tv tty include standard binding assays as described herein, for example, by Bmojre surface plastron tvsorumce (SPR) analysis using a B taco reJ ·" '2000 SPR instrument (Biaeore ΛΒ. Uppsala, Sweden t. or trow cy tot nets ic techniques.

As used hosem, "isotype s\s itching" refers to the phenomenon by which the class, or isotype, of an antibody changes from one hi class to one of the other Is classes.

Ah used herein, '‘nonswuched isotype5' refers to die isotypic class of heavy chain tbar is produced a lien no tsotype switching has taken place: the OH gene encoding this nonva itched isotype is typically the fust CH gene immediately downstream front the functional he rearranged YDj sene isotype switching has been classified as classical or nonclassical tsotype switching Cla-stcal isotype switching occurs by recombination events which involve a! least one switch sequence region tn the transacne. Non-classical tsotype switch!tty may occur by. for example, homologous recombination between human 0., mid human Τ,μ (d-associated dele cow. Alternative non-classical switching ntechamams,, such as intcnrsnsgenc and'or inierchromoso-nal recornbittatiott. among others, may occur and effectuate isotype switching.

As used herein, the term "switch sequence"io:.thoSe-l>|^iAst^ueftees· responslble ior switch recombination. Λ "switch donor" wvwcnce, typically a μ switch region, will he 5’ (/.e., upstreams of the construe! tcgmtt to be deleted dtinna flu- svvhch recombination. The "switch acceptor1' region will hr between the rows true: region io be deleted and the replacement constant region μ .y . γ. th etc. I, As there is no specific site where recombination always occurs, the final sene sequence will typically not be predictable from the e on struct...

As used herein, '"glycovyhtiost pattern" ts defined as -he pattern of carbohydrate units that arc covsIeniU attached to a protein, mmw specifically to an imminmgiobulm protein. A glycosylaiion pattern oi a heterologous anybody can be charactermed as being subswmiady «imisur to giycosylabon pattern- which occur natmally on ami bodies produced by dm species of the nonhunum trtntsgemc an tomb when one uf ordinary skill in the an would -ver-gm/e dm aheswykuion pattern of dm hetofologou- antibody us being more similar ?o said panmrn . a giycosykntoo in dm pmmos of she oonhutrsun niutscenle animal than to she specie- llvm which dm (11 amw'' of Urn warn mom won' deroed. 1' he semi “n.nm.dly--:.-cemTiog'' as used km mm- ;m applied to an : fame? re tors ro ihe f:.mi that an object can he found w nature, dor example. a pTypephdc m polynucleotide sequence that m present in act nr-mmsm uncluding slmsest that can be iso bred bout a soured in nature and which has not items nttem tonally rnochdcd h> man m the laboratory j<? patbr&amp;tly-· occurrmg.

The term “rearranged’' as tmed herein refers t<.> a configuration of a heavy chain or light chant hnmunogiobuiin locus wherein a V sea mem ts positioned immediately sdiaeent to a DJ or j segment in a ec-mdruinfion encoding essentially a complete V't! >_n V· domain, respectively. A tvarranged immunoglobulin grate tocus can be identified by comparison to yemilinc DMA: a rearranged locus svill have at least one recombined tiepinrnes fnonan mr homolopy elcrneηi.

Ihe. term 'Tnreas'nmped ' or "gems line configuration' ‘ a« used her rein in refetesme to a V seamen* refers to the configuration wherein rhe V segment ts not recombined so as to be immediately adjacent to a D or .1 segment.

The term "nucleic t« ;d molecule."' as used herein. i\ mtendbd . molecules and R IRA molecules. Λ nucleic acid molecule may be single-stranded or doublestranded, hut preicntbly ts double-stranded DMA.

Ihe term, “isolated nucleic acid molecule." as used he mm in reterenee to nucleic acids encoding antibodies or antibody portions wye . VT. Vt. CDR.Vt that bund to ΠΧ'Τ is intended to refer m a nucleic acid molecule in which tbc nucleotide sequences ettcodtng the mm body or antibody portiort arc free ot other nucleotide sequences encoding antibodies -w antibody pontons that bind antigens other than ( 4X17. which othe; sequences, may naturally fkml the nucleic acid in human genomic DNΛ. for wtmple SEQ ID NOs; 35 and .i[. 4" and Mb id'1! and l(w\ So and bb. S3 and 05, y5 and ,?.d. 71 ansi 77 correspond, respectively, to the nucleotide sequencer comprising ihe Iteavy chant t VT and light chain Vj} variable mginrts of am--CD27 antibody monoclonal antibodies I Ed, \ MS, 5H1 7, 3Λ !(k 202. 70b 3H8. and 105.

Thi? pirsent urn-mum abe encompassur, "eo men a? wo soooener' n mo fir,mens" of She sequences set u>rih htShQ Π) \(K: 5-1 12. ? >\. mwk\mde and ammo add sequrui.e nmdhli.athms v- Such dp um ado-cate the bmdmc oi the umb-xH ..-nr oded by {he nucleoside sequence or containing the enure> acid si-queuee so she midgcm. Such conserv anw sequence muds fees ;em-· include cem-es s am·.e aucieuiu.Se and sn'un· · ac-d s-ihsiifieiuiv·. as well an. nucleotide and ammo ;e.ni addition- and octet mm,. fm- es ample, modifications can he mnobueed I mo ff.Q H’> \O-a 9.1 id b\ ssassdas'd technique^ known in • be art - uch a-. ~.;Se-di seeled nadaccoe-m: and IkTR-niediased mutagenesis. ('on sew at we run mo icid sui-.0111010114. Sac hide one-' an which dm amine m.ai residue k replaced with an •amine· &amp;&amp;|d residue ftaving a similar side chain, Families of imtino acid t^sidues: bssnng •dm war -ads chains rune been defined in die no These funnies iutdude amino acid-' with h;e'Si. side debus -fee·- fywma arginine, histidmei, acidic side chains (my aspanic acid, gltihunk aeidf uncharged polar side chains use,, glvcine. nspar.tgine. ghmsmme, serine, threonine, wiOs-ne. cysteine, tryptophan nonpolar side chains (c.w- aia-drse, val-ne. leu>. me, Ssu Seise me. psoihie, phewylal an me, melfnomsies. beta branched side chains -.e.y . shim-nme, valine. isoiendrse') and arm runic side chains f- y.. tyrosine. phenylalanine, tryptophan, hisddini.n. Thus, a predicted nouesseuhal amino acid wsdee m a human ;mtoCP27 antibody is preferably replaced with another amino acid residue from the same side chain fund}. Methods of identifying. rmcieodde and amino acid conservalive substitutions winch do nor eliminate antigen binding are well-known in the an {see. e.y- Bmmmeh n a!» fthu hem. 52: \ ISO-1 IB7 (.1995f Kobttyasbi 0/ 0/. Protein Prut. I.?.{10Ί:8?9-8&amp;4· i !!Wi: and Burk', m ah Pw<\ N<ul. Λ cod. Sn. VSA Ο-ίι-ΐ 1 2--117 ·: 1997)1

Alternatively. in anotbej'embodiment, mmations can he unreduced randomly along all or pan of an nntl-CD2~ andbods coding sequence, such as by saturation mutagenesis, and the resulting modified ami CD./ / ami bodies can be screened for binding •activity..

For nucleic acids, the term 'Tebsmofd homology'' indicates that hvo nucleic acids, or designated requestors {hereof when oprirnady aligned and compared, arc ideniieah with appropriate nucleotide Insertions or deletions, in at least about BOB· of the nucleotides, usually at least about 909)- to vyff-. and more preferably at least about OBO- 10 99.59;. rtf the neelem'idos. Abe-mati\els. substantial homology exists when the segments will hybridize lit idea selective iw brkheadon conditions, to the complement of the strand.

The percent identity between two sequences 55 a function of the ouniber ot identical position1; shared by the sequences if;··.. % homology ~ # of identical posttimis/Knal ·# ot position'; \ 1 GOT taking into account rise member of gape, and the length of each gap. which need to be introduced tor optimal alignment of the two sequences. The comparison of sequences and determines bon of percent identhy between two sequences can be accomplished using a mathematical algorithm, as described in the none mm ting examples nelow:.

The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at http://www.gcg.com), using a NWSaapdna.CMP matrix and a gap weight of 40, 50. 60, 70, or SO and a length weight of I. 2, 3 1. 5, or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of II Meyers mid W. Miller (OABIOS, 4;j 1-17 {1 vSO'jx which has been Incorporated into the .ALIGN program (version 2 0;. using a PAM i 20 weight residue table, a, gap length penalty ot 12 and a gap penalty of 4 in add!Hon. the percent identity between two amino acid sequences can be deteimincd using the Neediernnn and Wunsch t./. A/m. Idol. {48):444-453 ; 1970)) algorithm which bus been incorporated into the GAP program In the GCG software package (available at hup;//ww w.gen.comb using either a Blossom 62 matrix or a ΡΛΜ250 matrix and a gap weight of 16, 14, 12, 10, 8. 6, or 4 and a length weight of I. 2. -1, 5. or 6.

The nucleic add and protein sequences of the present intention cun further be used as a "query sequence" to perform a search against public databases ίο, for example, identify related sequences. Such searches can be performed using rhe N BLAST and XBLAST programs (version 2.0) of Ahsdn.il, c? us. {i960) ,/, TAG Awe. 215:406 1(1. BLAST nucleotide searches can be performed with the XBLA.ST program score - 10(), wordlenglh ~ I 2 to obtain nucleotide sequences homologous to the nucleic acid molecules of the invention. BLAST protein smirches can be performed with the XBLAST program, scow* - 50, wordlengih - 3 to obtain ammo add sequences homologous to the proton:·mdlsculsi·..of-dlls. invention. To obtain gapped alignments lor comp.uison purpoew. (Tapped BLAST mm be nliliaed us described in .AHsehui ,-/ ?,</.. ι lv\>?i NucU ic Acid* (A s. 25{ 171:5569 5402. When ntilijriua BLAST and Gapped BLAST pro ami ns, the default parameters of the respective programs t'e.g.. XBLAST and N'BLAST* can be used See btifW/wwwnscbi.nlni.mh am.,

The nueide acids may be present m whole veils, ui a ceH lysate, or m a. partially purified or substantially pure form, A nucleic add is '‘isolated" or "rendered ,w baton really pun;" when pun-led away from oilier cellular components or other centa-mihaiils, qiihsr -oetjulsr. :&amp;£ίά$ difrsMas,: fey:.standdrd ^ebi$q:u%' iricltrdhig alkallne/SDS treatment.00 binding, column eh roman ygmphy . agarow: get eseeirophoicsb and others well know*; in the an, .%>*·. F\ AusuheL e/nd,, eo. Correm Protocols in Moieeular Biology. Greene Publishing and Wiley Inters:, lenee, New York ; 1987).

The nucleic acid compositions of die present invention, whale often re a nance wqnerwe useept for modified restriction site", and die liken from either eDV.-V genomic os’ mixtures thereof may be mutated, in accordance with standard techniques to po)<. tde gene sequence·-·, For ending sequences, these imitations. may affect amino acid sequence as dobed ht particular,. DNA sequences substantially homologon-, to ot tierssed fsom uatbo V, l), j. eon want. switches and other such ;-cqueuces described hemm are t orhemplated where "derived" indicates that a sequence w idemics) or modified from a: rot he: sequence), A nucleic acid is "operably linked'' when it b placed into a functional re rations hip v. ith another uuelcec acid sequence. For instance, a promoter or enhancer is operably hnked to a coding sequence if it affects the transcription of the sequence. With respect to t; si; scrip dor; regulates y sequencer, operably linked mean - that the DNA sequences beuis linked am contiguous and. where nrcessuss so loin two protein coding regions, contiguous and in reading frame. Fos' switch sequences, operably linked indicates that the sequences are capable of effecting swatch recombination.

The term "vector.” as used herein, is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of rector I-i a ''plasmid/' which refers to a circular double stranded DNA loop into which additional DNA segment·' may he ligated. Another type of sectm e' a viral vector, wherein additional DNA sesmeuts may be iig.ued into the \ ma! venomr. Certain vectors am capable of auionomous replication in a host cell into which they arc mfivuncrd ti-.y., bacterial vectors having a bacterial origin of replication and epboma! mammalian vectors). Order vectors (rip. noivepisomal mammalian vectors; can be hueg-med into the genome of a host cell spon· introduction into the host cell, and thereby arc replicated along with ihe host genome. Moreover....certain vectors are capable of directing the expression of genes to w hich they are operatively linked Such vectors are referred to herein an "recombinant expression vccioi's 'icr simply, "expression vectors '} In eeuend, expression vectors ot utility m iccomhmani DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" may be used mte.wharoseahiy ,w the plasmid Is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors. such as v ira) veeto-s (e.g., replication defects ve. retroviruses, adtmovs ruses and adenu-assoenited viruses . which serve equivalent fut unions.

The term 'Yeco.rU.huaM host cell1' for simply ''host eelG). as used herein, is intended to refes to a cdI into which a recombinant expression vector has been inueduced. It should he understood that such tenn.s are inrended to refer nor only to the particular subject cell tint to the progeny of such a celt. Because certain mooifk aborts stay occur in succeeding cenemhuns due to eithes mu-aiimt or environmental influences, .vach progeny may not. tu fact, be identical to the parent celi, but are sol! included o chin the scope of the tern; ''host ceir as used heK'in, A', used heteiit. the let m '’antigen" tefers its asty natural or synthetic immunogenic substance, such ;ts a protein, peptide, or hapten Suitable antigens lor use. ut the presem invention (e y- hi a vaccine tn costshmatton v>Uh an antcCP27 antibody of the invention) include, for example, Infect ious disease antigens and tumor antigens, against vehscb protect; se or therapeutic ream one responses are desired, e.p., antigens expressed by a tumor cell or a pathogenic os gun Ism or infectious disease antigen:; b'or example, suitable antigens include snrnor-asxocuited ami gens tor the prevention or it cat met tf of cancers. Examples of nm;o;-associated antigens include, but are not llnuted to, sequences comprising ail or pan of the eminences of jahC’G. gp 100 or Erne) 17, HER.emeu. Ά“I t, stvsoihelhi, CBA. gp»!00. MARTI. TRP-2. mount A. NY ESO t, NV-8R-1. NY -CO-58. MY tgp250r idiots pc. MAGE 1, MAGE-3, ΥΙΑΟΠ-Αλ Tyn>sma*e. Iciotnetase. SSXd and MUG 1 arthnetts. rmd gertn cell dertved tin ten antigens. I it tnor associated antigen·* aim· mclndbfhe blood group antigens, lor example. I e”. Lem LeX. LeY. H-2, B-1, B d antigens.

Alternatively. snore that: one antigen cast be included nothin the anrmen-antibocy· constructs of She Invention. for example, a MAGE anti sen can be combined with other antigens such as rrtdlanip A, rymsmase, gad: gpifMl alongwith adjuvants such as GM-CSE or iEkl2vandJinked to an anti-A PC antibody.

Other suitable antigens include viral antigens for the prevetttion or tsestmetit of viral diseases. Examples oi viral antigens Include, but are not limited so, HIV-1 gag. HiV-I env, HIV-1 net, HBV .'surface or cose antigens}, HPV. PAS. HSV 1. HSV-2, p!7, ORfm and OR P.3 arsisgenv. Examples of bacterial antigens include, but -are not 1 nutted to. Toxopiasnui gondii or Ί rvprtm'ma pallhhat). I he antibody-bacterial antigen conjugates of the invention ear; be in the treatment us prevention of various bacterial diseases such as .Anthrax Botulism. Tetanus. Chlamydia. Cholera, Diphtheria, Lyme Disease. Syphilis and ?h beret t loses. Other suitable antigens from its feettons-.disease jpathopaiS, soplh.$sevirase:&amp; bacteria, parasites and fur-gs are disclosed helms .

Sequences of the foregoing antigens are wed known in the an. For example, an example of a VIAGL-3 eDNA sequence x; poo bind in US6.2V5.52b (Ludwig institute (or Cancer Research); ex amp Vs of NY-F.SG-l nucleic acid and protein sequences are provided in US 5.804.581 and V.% 6,000,255 t Lochs tg institute for Cancer .Research): examples of Rician- A nucieic acid and protein sequences xve provided in US 5..620,,886 and (bS 5.854 205 U.ndvUg Institute for Cancer Research); examples of N Y-BR i nucleic acid and preach; sequences are provided in US 0 774.226 and US 6ΛΗ 5,52.0 ; Ludwig Institute for Canoes Research > and example:·; of N Y-CO-58 nucleic acid and protein sequences are provided in WO 020009&amp;fc (Ludwig Institute tor Cancer Research); an example of an amino acid sequence for the IH5R-.?./ ten protein is available at GLNBANK® Accession No, ΛΑΛ58657' and a tiucleotidc sequence (mRNAi fen human esuvsnoembryemk .tvuigendike i i{.IA-l) is a variable at 6ΕΝΒΑΝΚΦ Accession No. NM„J)202lv.

An HPV antigen that may he used In the· compositions and the methods of the invention may .include, for example an HPV' id antigen, an HPV- 18 asm yen, an HPV-8i antigen, an HPV-33 antigen and/or an HPV-35 antigen; and is suitably an HPV'-lb antigen and/or HPV-18 antigen. A genome of HPV- lb is described in Virology, Ι45Π&amp;1- 185 (1985) and UNA sequences encoding HPV-18 are described in US Patent No, 5,840,806. the disclosures of which are irscorpotated by reference heu-ηη in their entirety. HPV- ip antigens te.g., seroreaetive regions of the El und/or r,2 proteins of UPV-lbi am described in US Patent No. 6,531.127, and HPV-IS antigens le,g.. aetoreae-tse regions of the U and/or 1.2 proteins of HPV i8i are -described m US Patent No. 5,84().306. the disclosures of * inch are incorporated by reference heroin. Similarly, « complete genome for HBV x. available at GF.NBANR® Accession No, NC..005977. the disclosure of which is incorporated herein.

The genome of HCY is described in European Patent Application No. 318 216, the disclosure of which is mcorpomted herein. PCT/US°0/0I3'48, weorposu-ed by reference itendn. discloses sequence information of denes of the HCV genome, amino acid sequences of HCV viral proteins and methods of making, and using such compositions tor HOY vaccines compnsittg HOV protem s and. peptides dedved them fmsii.

Antigenic peptides of proteins those containing T cell epitopes) can be identified ut a vassety of manners well known in me art. Rr example. T cell epitopes cart be predicted by analysing the sequence of the protein using web-based pmuieiive algorithms {BIMAS &amp; 8ΥΓΤΤ:ΤΓΠΓ· to generate potential MHC class ϊ and 11 binding peptides that match an internal database of 10.000 well characterized Mi-1C binding peptides previously defined hy CTLs. High scoring peptides can be tanked and selected "interesting0 on hie bards of high affinity to a given MHC molecule.

Another method for idem d ying antigenic peptides containing T cell epitopes m by dividing the protein into stun-overlapping peptides of desired length or overlapping peptides of desired lengths which can be produced wcombmamly. synthetically, or in certain ihnlted situations, by chemical cleavage of the protean .aid tested for inanumogenie properties, i.-.y,. eliciting sT cell response ip.e.. proliferation or lymphobme secretionh hi order to determine precise T cell epitopes of the protein by, for example, line mapping techniques, a peptide ha vans 7 cell stimulating activity arid thus comprisma at feast one T cell epitope, as determined by T cell biology techniques, can be modified by adds;ion or deletion of amino acid residues at either the amino orrarbosy terminus of the peptide and tested to determine a change in T eeb reactivity to the modified peptide, 11 two or more peptides which share an area ot overlap in the nance protein sequence are found to have human T cell stimulating activity as determined by T cell biology techniques, additional peptides can be produced comprising ail or a portion of such peptides and these additional peptides can be tested by a similar procedure. Following this technique., peptides am selected and produced recornbinataly or synthetically· Peptides are selected based on various factors, including the strength of the T cell response to the peptide «*·.?;. srinndatiop: index h The physical and chemical properties of these selected peptides hop., solubility, stability! can then be examined to deter! nine vvhethes the peptides are suitable for use in the? a pen tie compositions or whether the peptides require modification.

The ierm '\uninen presenting cell" or "APC" ts a ceil that displays foreign antigen complexed with MHC on its surface. Tamils recognise this complex using T-cel! receptor f T OR > Examples of A PCs include, hut are nor limned to. dendritic ceils (PCs·, peripheral blood mononuclear cell;: < PB-MO, monocytes (such as THP-1). B lytnpboi'lustoi.d cells (such as C1R.A2 I'MS Bd Cl .t und monocyte-derived dendritic.cells (DC.su .Some A he's iuiernali/e antigens either by phagocytoso or bs nrcepiof mediated em.iocytos-v Haamples of A PC receptors include, hut are not limited to O-type lectins, such as. the human Dendririe and Epithelial Cell 70b leveptor tCOi?!. and tbe. human macrophage mannose, receptor.

The term 'vamtge:t j»es<fM»tioft'' refers to the process by which APCs capture aatinem; and enables then recognition by T-cefH. as ;.t component of a π MHC-1 and/or MHCVli conjugate. "'MHC molecules’’ include tun types of molecules. MHC class ( ami MHC class 11. MHC' class l molecules present ants yen to specific CD is t T cells and M HC clast; Π. moimmies present antigen to specific CD4+-1’ cells Antigens delivered exogenously so APCs are processed primarily for association with MHC class II. In contrast. antigens delivered endogenously to AK's arc processed primarily tor arsociation with Ml \C class I.

As used herein, site terra Arnmurmstmn.ilniory agent" includes hut Is not 1 ironed to compounds capable of stimulating A PCs, such as DCs and macrophages. For example, suitable irmmnmsihnulatory agents lor use In the present hr-onbon are capable ot stimulating APCs so thus the mates as ion process of the API's is aeeelenued, the proliferation of Apt's is increased, a-td/or the mcrultment or release of co-stimulatory molecules ο·'.η·< CD80 CFCC ICAM-I, MiTC molecules and CCR7} and pro-Inflammatory cytokines (me.. Π. A is. II. A. 1(.-12. 11.-15, and IFN rs is upregulated. Stumble immui-oxtimuiatory agents arc also capable oi nx rests; a a 1 cell proliferation. Such im-nnaostimolatory agents include, but are not be limited m, CD40 ligand: FLT 3 ligand; cytokines, such as IFN-m Π'Ν-jC IFN-y and if.-3; cuhats-stunulnting Psctoi's. such as O-C'SF (granulocyte colony-stimulating Pteturs and CAI-C.SlHgrainilocxte-ntactVjphawc colony-stimulating factorK art anti'CTl.A-4 antibody, anli PD I antibody, and-4 IBB antibody, or antl-OX-40 antibody. LPS I endotoxin); ssRC A: dsRN’A. Bacilte Calmette-Guam (BCD p (..evamisole hydrochloride: and intravenous immune glob-dsns. Id one etnlmdimem an srnsrmnosiimuteiory span? may be a Toll -like Receptor PTI R; agonist. For example the inunimostlmulatory agent maybe a TLR3 agonist such as d< mole rbanded tnosinetcx-toxtoc polynucleotide {Poly 1:C, fos' example awn Abie as AmpItgenTM Faun llemispherx Biphmma. PA, US or Pojy ICtLC froot Oncovir) or Poly ATI; a T1..R4 agontst such as monopho«phoiyl lipid A (MRLt or RC-539 ifo; example as asaiiablo front CSC, His,)', a II R5 ago ana such as fbgelbu; a T1.R~ or TLRS agents’! such as an hmdazoqumoiine TLR? ut CT..R S agonist, for example tmmuimod teg AidamTMi or ieskpuiTtod and related irmdaaoquinohne agents ttor example ;m available front aM Corporation}; or a Ί LR b agonist such as a dermynoekxuide \x uh ««methylated CpG nsatrfe {so-called “C'pGs”, for example as av an able itmm Coles PfamuareouvaP. Λ prefened immunosthutdatory agent is a '1 LR3 agonist, pecferubly Poly l;C. Such irntnunostinvulatory agents ruav be administered simubaneonsiy separately or sequentluiU v-·HIt the antibodies cttBiStfacls of iW- preseat iivveivao» asd may akdte physically imfeed M the antibodies and constructs.

As used herein. the term "linked” refers to the association of two or more molecules'. The hrtkage can be covalent or non-covalent. The linkage also can be genetic {<#., recmnbmamly fused). Such linkages can be achieved using a wide variety oi ait teeognkcd techniques, such as chemical conjugation and recombinant protein production.

As used berbdh the ΜΑη :an.tigen;'^os§i:|>rescn{atipit''' refers to presentation of exogenous protein antigens to T cells via MHC class I and class Π molecules on A PCs.

As used herein, the terra "T cell-mediated response" refers to any 5espouse mediated by T cells, including efi ector Γ cells CO# cells; and helper T cells :< y., ••034* cell# T eel! mediated: mspopsek i behthe, fer ekample¥ T eel! eytoioatedy end; proliferation.

As used hetem, the terra "eytotos'ic T lymphocyte (CTi,s msponsr" refers to ku immune rmpons:.· induced by cyioiosic T cells, i' ll., responses arc mediated primarily by CD# f cells. \s used herein, the lean'. "iulht-hs" or' block.'” Osm, itivrrmg to inhibidon/bf x'kiug of binding of «'Ί.)70 t<;0')2? on cells i arc used interchangeably and encompass both partial and eompk.se mhihalun/l'kfchins. The Inhlbnioa/Olucking ol CO70 preferably reduces orairers the normal level or type of act# its that occurs 'ahen 01)70 binding occms without initibuton or blocking. Inhibition and blocking are also I mended to include any measurable decrease m Uv binding :Afina} m CD7o a licit in contact with ait antrCDo? cud body as eompmed 10 CD70 not in contact with an stur-GDa? antibody e.y.. inhibits bind I tig of CD70 by at least about I 0#, 1 5#. '’Off. 25#. 50#. .##, 40#. -15 A, 50¾. 5:51¾. 60¾. 65¾ 70#, 75%:, 80#, 85¾. 00¾. 95¾. 96¾ 97¾ 98¾ 99#. or 190#. hi a pief'erred embodiment, the anti ( ’1977 antibody inhibits binding of CD7Q by at least about 7(1¾. in another embodiment, the aou!-CD2? amuxxh Inhibits binding of CD70 by at kastJO®

As used herein, the term '‘inhibits growth" (e.g.. referring to cells; is .intended ίο include any measurable decrease in the growth of a cell, cm., the inhibition of growth of a cell bv at [east about 10#. 70¾, 30#, 40¾ PL#· 60#. 70#. 8it#, 900-, 99(3., or 100#.

The terms '"inducing an immune response” and 'enhancing an immune .reapeuse" are used interchangeably and refer the stimulation of an immune response Onm ei titer passive or adaptive: to a particular antigen. The terms ‘induce* as used 'with respect' to inducing CDC or ADCt' refer to the stinuiiaiton of parucuiar dime· coll killing mecbamssus, For example. in e<ne eniboohrnent. km amibi#lv lndm.es at least about 20, 2a, 3lk 35- -10,45, 50, 55, <n o0 5?· lysis s in i 'DC of CD2? expressing colls at a coneenlraiioti of dip g/rnk in a pa etc r rad embodunenh the antibody seduces at leak about 40 A lysis >, ja 0X5 of CD2 7 expressing ceils at a csutcesmation of lOpg/ml. its anothe; embodiment. the antibaidv induces at least about 20, 25, 50, 35, 40, 45, 50, 55. 00. PS., 70, 75, 30. or 35'X- lysis via ADCC {?,/.·,. «per he Axim of (75)27 e\pmx-<mg cell-, at a concentration of It fug/mi. In a pmierred embodiment, the antibody induce;» at least about 40 Ά lysF; vsa ADCC of CD27 exp-easing celts at a uoneemrtuion of I0pg/mi.

Tiie terms 'kseak” "tmaiing/' and "treatment,' as used herein, refer to therapeutic of preventative measuses described herein The methods of "treatment” employ administration so a subject, in need of such treatment. a human antibody oi the present invention, for example, a subject m steed of an enhanced immune response against a particular antigen or a subject u In> intimately may acquire such a disorder, in order to present, cure, del as, mduee the severity of. os' asneuorate one or more symptoms oi the disorder of recmTirtg #scTdor, at in order Tp: prolong the survivaf Of a subject beymtd that expected its the absence of such treatment.

The term "effect ive dose” or ‘'effective dosage'' is defined as an amount suit idem to achieve or at least partially achieve the deseed effect, rise term "therapeutically efleetive dose"1 is del sued us an amount sufficient to cure or at least partially arrest, the d-mase and Its cum plications m a patient already suffering irons the disease. Amounts effective for this use will depend upon the sexes sty of the disorder being treated and the general state ot the patient’s own mi mo no system.

The testsi ''pattern'' includes human and other mammalian subjects that receive either prophylactic or therapeutic tretit.me.nt.

As tr\ed herein, the term "subject*' includes assy human or non-human animal l:or example, the methods and compositions of the present invention cats he used to treat a subject with an immune disorder. The term "rson-htsmtus animrti" includes all senebratex, e g., mammals ami non-mammals. xs.ich as non human primates, sheep, dog, cow. chickens, amphibians. septiles, eic.

Various as pee ls of:the ihvehdort: are described: in farther detail In the folio waqg. subsections. 1, Prodncm\fi -?f Ai:d 1 a · IC;-jcyCj>27

Til- present invention encompasses antibodies. e.g., lulls hunem antibodies, thus bind CD;?.?, we. human (’027. iTemplars snmioclorsal anisbodies that bind C.D2? include IP*!. 1HT 3HI2. C\10. 2C.2. cOim ΛΗ8, and IGN VhinockmrU run;babies c.Cme inveiifion can be produced using a varies y oi known technique:», such as the standard somatic ceil hybrid waiesn techsslqtse described by Kohler and Mi l stem. Nature 25b: 40:5 1075 c Λ|Coup!; somatic tell hybridmaUmi procedure^ are preferred, in princspk. oldertechnique-; for producing monoclonal atntbodies also can he employed. e.y v usd or oncogenic rnuud'ornuibon oi H lymphocytes, phage display seehumue nsuia binaries of been van antibody genes..

Accordingly, m one emhodinsem. a hybridimia meshod is used for producing an endliody rhe; Cuds human CD27 In this method, a mouse or other appropriate host animal can be immsmiaeb with a -ontahie antigen in order to elkss lymphs-evies,..that produce or arc capable of probocmg antibodies thus o, id specifically bind to the antigen used for snsmunkatlon. Alternatively, lymphocytes rna> be immunised m iCe. 1 .ynsphooy urn can men be fused mi lb myeloma eel 1-. using a wihablc fusing agent, .-tseh as polyediylcnc glycol, to iorvu a hybrldorna cell {Coding. Monoclonal Antibodies: Principle'- and fine tier.·, ρρ.59-KM (Academic Press, IPICoi. Cubure mefimm in which hybridous:.: eel;-. are mow mg is assayed tor production ;;f nsonocionai antibodies directed a gas no die ami pen. .After Mmfidosna cells are identified that produce antibodies ol the deseed specificity, affinity, and/or acti- ns. the clones may be subcloncd by fimeme dihmoo procedures and grow o by siandurd me: hods iGoding. Monoclonal Asitshodics.iVhsosnles and bracficc. pp, pfi-lOo {Academic Press, IS>spfis, Suitable culture media for lie:- purpose mcinde.. for example. ID-MEM or RPVH-1(:-10 nseilium. In addition, rhe hyhrsdorna cells may bo groom b; vivo as a-a. me s isunors is; an ashmal. The mcmoekmal antibodies secreted by she subebanw c a n he separated from toe cultnro mediums, usches fh.nd. or serum by conventional uu.mu.noglcbohn pssnlscatson procedures ssseb as. for example, piotcin .A-Seph;.tsose. hydsom iuparue chroiTKtiograpby, gel eleets'ophoresis. dialysis, or affinity chuomatemsuphy. in another embodiment, antibodies and antibody portion-, this; bind buss· in OD27 can he isolated isx>m antibody phase libraries generated usine Use techniques desetibed its. for example. MeCaffmty ct mb. A'nmre. M8:552-554 GS-Kn, Ciackr.on m mb. Nature. .152:624 (Mb ti^ik Marks w a!.../. Moi Biol.. 222:581-507 s IPOhattd Hoetm oi l2005t Nature iliotechoob$v 23, 344-3-18 : I f $ Patent Nos. 5,225,-IOv; 5,403.484: and 5,5? 1.698 to

Ladner e/η/.: IKS. Patent Nos. 5.427.908 and 5,58(),717 to Dowers d/,; I'-S Patent Nos. 5.909JOS and 6,172.19“ to McCat’fVftv a al.: and IJ.S, Patent Nos. 5,885.793; 6.52.1,404; 6,544.731: 0.555.513: 6,582.915 and 6.593.981 to Griffiths (} ¢//., Additionally, production, of bid; affinity ;ηM range) human antibodies by chain shuiTIme ?>larks < / id.. 10:7? 9- 783 C 7992)), as: well as combirralorM Infection and; irt mm recombfuat ton as a strategy lor constructing very large phage libraries tWan-rhnuse et A';/;·, At his /As.. 21:2365 -7306 i Ife-J?t may also be used.

In a particular embodiment, the antibody that hi nos human CDs 7 is produced usuis* the phage ah splay wchrmp.ie described by Hoot a ¢//., supra. This technique involves; the generation ef a bmnan bait binary having a unique combination of immunoglobulin sequences Isolated iron; human donors and having synthetic diversify in the. heavy-chain ODRs is generated. The library is then screened for labs that bind to ho man CP57.

Theqreferred animal system for generating hybrklomas whsch produce an Obi-:.5-.-s of the invention D me murine system. Hybrirfoma production in the mouse is o laioysn in the art. including immunization protocols and techniques (=? isolating and 1 using immunized spinner ytes.

In one embodiment, antibodies directed against (51,)5 7 are generated using trausaemeor tmnschromosorrud mice carrying parts of the human Immune system rather thaiii the mouse system. In one embodiment, the invention employs transgenic mice, refeneu to herein a;- "HuMAh mice*' which contain a human imnumoglohulin gene min I loci that encodes onrearmiigoo huniuu heavy {μ and y) and κ light chain mununoglobuiin sequences, together with, targeted mutations that inactivate the endogenous μ. and κ chain loci iLonberg. N. ¢-/ ¢// {1004'? Kobo·? 30&amp;6474); $56-859) Accordingly, the mice exhibit reduced expression of mouse 1»M or K, and in response h« immunization, the Introduced human heavy and light chan: U'ansgenev undergo class sw itching and somatic mu tat tun lo generate high al Unity human igGk monoclonal antibodies (1.on berg. N, ¢-/ at ί 1994 b supra: reviewed ;n (.onberg. M, (1994) Handbook of fixpirbniana! Pharmacology \ 13:5-9-101: Lr.-u.0erg. A. and Huszar. f) f 199.5) haem. Rev. hvaamoi. Vol. 13: 65-53, and Harding, F. and Lon berg. N i 1995) Atm. Ac Y. Arad Sot 764D36-546) The preparation of HoMAb mice Is described tn detail in Section II below and in Taylor, 1.,.;:i of (1552) Nmifit ArhH MesnircP 50:658767:95: Chen. .1, «/ al. i 1993,) Pikoaodoao:! Ipimaaology 5; 695-656; Tumi ion ¢-/ al i 1993) fbtfe. Nail, Aadd* Sci USA 99;3720-3729y Choi m ol (1993) Mitere Qemuds:4:1 I T-123; Chen. .1. - v al. {169)? j-'MRO./. 12: B21 -330: Tuadion et al. {1994)./. ImmimoL 152:59; 2- 2020; Lost berg ¢/ (0204) Naiure 368(6474); 850-850; Lonbesg, N. (1204) Ηωκίί*ο*Λ of'·

ExpmmmiaiPkarmaeMogy ! 13:49-191; tAylbr,.:L.:gi ol (>; 572-501; Lonberg N. and Hus/.ar, D, (,1995: 6//0,0/. AVt. iiomioioi. Vo!, 13: 03-23; 1 (a-dhtg, F, ana Lemberg, N. 0905) Awk A'. K A^ad -Vs 76-f 536-5-16; Fishwild. D. et at {1996) Nature Biou-'hudni'y h-h 845-851. See Anther. lob. Patent No1·:, 5,545,800; 5.509,825; 5.625.126: 5,655,425; 5.789.050; 5,877,307; 5 60i .016: 5,8 5 -UIB; 5,874,300; and 5.770 429' all to Losibrrg and Kay. and (.IanPham; htternath.mal; U.S. Patent No. 5,545,8()7 ;o Snrani < / .·;/.: International Pubhcanon Non. WO 98/.M88-4 published on June 11; 0398, WO 94/26585, published November 10. 1994; WO 9Vi 227, published June 24, 1993; W(> 92/22645, published December 28, 1992; WO 92/03918. published March 19. 1992, jmm tmziHions

Id aenerafr 046, ho man antibodies to <5D27, nane-genie or traoschamtoxornai ;mlc^'CiOMal«|ilg%Utnan smmuooglubuim gesics (e.y , HCol2. HOo3 or KM mice: can be irnrmmized with a purified or enriched preparation of vho CD27 anti sen and/or cells expressing CD7.7. as described, for example, by Lcnbe-rg of oL (199-1) N-tfure )68(0474): 846-859; Hshvcdd a aL (19%) Kuinre Bmt* hm/togy 14' 8-15-851 astdAVC} 98/24884. As described herein. HuMAb mice are immunized eithe· with reconmhnut? OD27 pro tents or cell fines expressing CD27 as immunogens. Alternatively, mice can he immunized svnh DNA eneodmg human C027, preferably, the mice veil! be 6-16 weeks of age upon rise .fir st infusion, For example, a pndiied or ondehed preparaban {509 pg) of thb r'ecem'binadt GD27 antigen cat; be used to annum lev the HuMAb mice intraperiioueally. In the event that immunizations u-ini; a purified or em idsed preparation or the CD2'· antigen do not result in antibodies, mice era; also be immumreu with cells expressing 0D27. < a ceil one, to promote immune responses f.sempl.ir\ ceil Hues include CD27- overc xpressmg stable OHO and Ruji edi Itnes.

Curnultrive experience u. ith v arsons antigesis Isas shown that ihc HuMAb transgenic mice re-pond best when initially immunized inirupentosteally UP: or subcutaneously 5 SO: with antigen is: tomplcte breund’s adiuvara. followed by every or ha week IP/SC immunization- (up to a total of l(o with antigen ht mcompkue Freund's adjuvant. The immune : espouse cat; hr men Tv.· red over the cour-e of the immunization p-moeol v- uh pkwrna samples ?·..-mg obtained by ren''-orhhai bleeds. The plasma Cat: be screened by Lie6A <us described below?, and mice wish .sufficient titers ofanti-OD27 human immunoglobulin can be used for fusions. Mice eon be boosted intravenously with antigen 3 days before sacrifice and removal of die spleen.

ikneoOio!! ofH'.'ht'irfonttis Prf/dm'h<i> Mown lemtt! AtaiJxidk's to i.7 EV

To generate hybndoroas producing monoclonal smt bodies to CD;?.?., splenocyiex and lymph node ceils Irani immunised mice can be isolated and fused to an appropriate mnnoriakzed cell line, such as a mouse myeloma «.ell line. The resulting liybridomas can then be screened for the production of antigen-specific antibodies. For example, single cell suspensions of splenic lymphocytes from immunized mice can be fused to SP2/0-AgB.013 non secreting mouse myeloma cells i.ATCC. CRI. 1580) with 10'? PEG f\\7vs. Cells can be pbted at approximately 1 \ 10 in flat bottom miccotiter plate followed by a two week incubation in selective medium containing besides usual reagents 1.0¾ fetal (/tone Serum. ό· KKt ongen hyhridomu cloning factor iiGBAd and IX HAT (Sigma), After approximately two weeks, cells ears be cultured in medium m which the HAT is replaced w 1th HT. Individual wells can then be screened by ELISA for human auti-OD27 monoclonal isM and IgG antibodies, or for binding to the surluce of celts expressing (31727. ¢- y., a CHO sen I line expressing CD? 7, by PUS A (bi u orem e nc e - 1 I eked immunosorbent asms?. Once e.xteuxwe hyhridoma growth occurs, medium can be ol.wer.ed usually after 10-14 days. The antibodg secreting hybridomas can be repiated. screened again, and if AH positive Ion IgG, anti-CDc? monoclonal antibodies can be subcloned at least twice by limiting dilution. The stable subdones cun then be cultured iti vitro to generate antibody in tissue culture medium fe-ohafacton-zattofe.

Ummmtkm rtf' IkmisjotooMao Producing MonmdoooidinSimdmt to CB2d

Antibodies of the invention also can be produced in a host cell trunsfectoma using, for example, a combination of recombinant DN A techniques and gene transfection methods as is well known in the art {Morrison. S. (1989) Science .7.29; 1202). for example, in one embodiment the genets t of Interest, e.y.. human antibody g<mes, can be ligated into an expression vector such us u eukaryotic expression plasmid such as used by GS ye tie expression system disclosed in WO B7/04402, WO 89/(:1030 and EE 338 841 or othci expression systems well known in the art. The purified plasmid with the cloned antibody genes can be introduced in eukaryotic host cells such as CHO-cells m NSO-cdK or alternatively other eukuiyobc ceils like a plant derived culls, fungi or yeast cells The method used to introduce these genes cot·Id be methods described in the art such as elects operation. Hpolociine, hpofecmmme or othe·. Λ tier i Producing these antibody genes in the host cells, cells expressing the antibody can be idem died and selected. These cells represent the traosleciomas which can then be amplified for their expression level and unsealed to; pasclucc-anyfeodiey. ReeoniMtmtit: antibodies eat!fed isolated and purified frod*. these culimri sc: pevnata nix and/or cel I s.

Alternatively these cloned antibody genes can he expressed in other t.mp:essmn systems such as d < oh or in complete organist ns or cart be synthetically expressed. lode &amp;f· 1¾IMid An<(hi>,·:h: 5/syaon-:co to- Express kittl(Φ ΑμΦμΙΙ^Χ:

ArtHbi-dt;". interna vs tilt tat yet antmero- pmdommanits tm'mtgh ammo -w ;d residues that ate loaded In the s:\ I no a v) .aid light chant emisplemcntaru·, dsterrntnnte regions (CPR-m pot dm. mason, the ammo acid 'vqnerit cs w. qhm ('HRs ore mmedtvorse between individual antibodies than .sequences outside of (T>Rs, Because CDR sequences arc responsible for most antibody -antigeu interactions, it Is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies, by constructing -expression vectors that include CDR sequences thorn the specific naturally occurring antibody grafted onto framework sequences from u different antibody with dirtVtem propedle® fsee, etf,, Mfechmunth !99B?.i^tixr®332i323~32?; Jotiesi R, etidk.· 19.8.(1-

Natan: 3a 1:523-5.7.5: and Queen, (/. <·/ οι.. 1989, Proa. Nad. Arad. Sct\ U.S.A, 89;! 0029-10(133,!. Such fmme work sequences can be obtained from puiihc DMA database', that include germ line antibody gene sequences. Those germhoe sequences will differ from mature antibody gene sequences because they will no? include completely nmemMed carbide genres winch urn formed by Y{D)3 joining during B cell matt; niton. (ΐ emu sue gene sequences will also differ from the seqitences of s high affinity secondary repertoire antibody m individual evenly across the variable seghnr For example.. somatic mutations are relatively httiequem in the .mthto-uenninni portion of framework ·ο|?1οη. For example, somatic mutations are tekrlvely mfrequent Ini the ammo .termmal X and in the carhoxy- tevrowal portion of framework region 4. F'urthetmom. many somanc mutations do not spmidecmiiy alter the binding properties of the antibody. Fur this reason, u u-; not necessary ίο obtain the entire DNA sequence of a particular antibody lit order to recreate an intact ret onsbloant antibody having binding ptnperties similar to theme of the original antibody ;mee piChC’SRC'Ofd.C filed - m Man,, h 1.:. mbXo pun Cl heavy and light chain sequence spanmng the C'DR regh-ns Is Typically sufficient lot this purpose. The partial 'vqnence is used to determine which aernhnie variable and joining gene segments cuiinibuied to die tecoosbsncd antibods \anabk.· genes The pernillfis· sequence l·; then used ίο ltd rn ini'·sine put lions of the % enable ughms i leas y and tight sheet 1 cadet sequencer arc e lensed during protein maturation and do not cwnrlbne to the properties of die final antibody io add muonic sequence·:. cloned ct)N \ sequences can be combined v. hh synthetic oh goo ae m*‘Tide-. by ligatmn or PCk umptlfkoden. AUernaTtveK. die cm Ire vaslabie my tot; can be vyntheaeed as a set of .short overlapping. oligonucleotides and eon dimed by PCR mnpufmabon to create an entirely synthetic variable reemm done. Thi- pn-ee-o has certain ad mm my os such as elimination or tnelusion or pmlicuhn rev Inchon sires, or epsunmotmn ·..? narrlenlar e-.bons.

The nndeoude sequences o! hem y and light chain transcripts bom a hvbndoma .ere used to de-lgn an ''verlapping set <b xy rehe:;c ohm‘nuciooudos t-o create synthetic ¥ sequences with idctdleai amino...acid eodliti capaeittes; ;af the :q#b.ra! sequences, The synthetic heavy and kappa chain sequences can dh'iet from the natural sequenees in three omyo -a rings o-f repeated nudeotide bases am mtcmmtcd κι facilitate oligonucleotide xvnThes··. and PCR amplification; opbmnJ irons his mm initiation sites a; e incorporated according to KoeakA rides tfwak. 1991. j. Biol. Chen;. 2hie 1 0867 |9k7(.n; and.

Hhtdiil .sues am etsgmeered ops;res;n of the translation initiation sites. l%r bath the heav) and light chain variable reborn-, the optimised codmgj,and corresponding non-t.Oi.hny. strand sequences arc broken down tmo 31] - eO nucleotide approximately the midpoint of the corresponding nomeoding oligonucleotide. Thns. for each chain, the oligonucleotides can be assembled into ok. or tupping double stranded sets that span segments of 1 w) - 400 nucleoSidcs. The pools are then used as templates tit produce PCR amplification products of PH) 400 nucleotides. Typically. a single variable teuton oligonuekolide set mill be broken doom tmo two pools which mm separately uiuninler! to generate two overlapping PCR product'. These overlapping products are then combined by PCR usuphimohon t-.< form the complete variable region, (t nitty also he desirable to inf. In do an overlapping ftautncnl ol the heavy or light chain constant region itnciudtng the Bo si site ol the kappa tight chain, os the Age! site tf the gamma heavy chant) in the PCR. amplification to gettetale ftaginctro that can easily be d-med into the exptessiou vectot constructs.

The ivceswi-acted here, \ and ugh? enam vat table regions me then cemMnrd v. uh cloned promote;. leader sequence ΐΓηη%<υη·η unPation. leader sequence, rus-suno reason, 3' nsnraiwiated, polyadens baton, and υηϊ'οοαρρ·,·;: teortmutton, oapn'n-..es ίο 1’ ,-rm expu'cenon set tor constructs The hu;n \ are! bobs chain r\pses - no; conernets cm: he ec-tnhmed ire'·.· a m-gb- eons ce-trnusieeted. 'Oijulls men-dected. or scraninch baasloctca : ni- > host cell' winch arc Shoe fused b< hum a h--st -, el I express Inc b-en chad tv, IM ivrneJv ler eve In construction ot expitssiou sestets were oom-tmeted no dut l'X"R amplified V neon·.} and \ Lappa ha hr chain oDNA sequences e--nld be used m : neon struct eompkne hcu\ y and Usb· ch.un nan h gene-· Them pbantids can be used to express completely human iaCtk'or IgD χantibodies H4K htsmau and chimeric annhodics of the tm sent no/emiun aK.· uieiotir iaf 12, hhla. (el , IgA, IgM end Ld> mmix'die-' dunlins plan-msis uun be eosn.triich'd for'.'Xprr.ss-en ot other heu\ y ;. hum ssotypox o- Tea CsptCOon ><f ,umbo-rim-; oinprivmsi non lain light chums

Huts, ;: 5 nuoiltct a\ pees -,-f-he invention. 'fruetural fVjona of muM Id a A atnmodtos oi foe hounbon ate toml ιο create slinctutuiSy related aoh CD7': m-hAme- ihai otam a I least mte luncponu! no-party ot the atmlm-M·' M ihe -mention, sums as, fm example, t1min bits u .y.. i. ore pi etc I) or prat-ally blocks'- Madina ot CD 70 to Π.Χ! / expremm:.'. cells by at least about 70%- {e.y,; by at least about 70%, or by at least 70%·, at an ami body concontrariors of 10 py/nut; t aS bmda jo human ΡΠ77 with an equilibrium dissociation constant Kd ol KM M or learnot' sdiernypody. an equilibrium uasoemiton constant Ka of 10*'' M * or greater i Ci induces at least about 30% complement mediated cytotoxicity fCDC) of CD27 expressing cell·; at a concentration of 10 ttg/ml to- to duces at least 3(07, oral least about 40% or at least 40% CPC of CD',": ” expressing cells at a concent ration of 10 μη./: a!): i -l) induces at least about 3^% spec:0c ADCXAmedhted lysis of CD27 expressing cells at a concentration of 10 ag/Am tor induces at least 30%·, or at (east about 40%' or at least 41)% speett to A DCC-rued: a ted A so; of C" 1.477 esq.;;ess mg cells at a cotu.entrat.mn of K) ug/.mlg bD prevents or inhibits the growth of CP27-expressing, tn-tsor cells in a xenograft model t-e.y., reduces motor si/c In wvere con; b joed :rnnu.utodei tcitncy (SildD; mite by at least about 5037 70-days post tumor cell inoculation to t A'.o at 0.5ms ip on ,u lete-t b days p (6- im.b.K:es or enhances antigen specific btunnut* res^.:rstses when combined with a vaccine or other antigen: (?'ϊ induces or enhance π In particularbut not limited to TBi immune responses: (St induces or enhances T-cdl activity, in particular but not limited to specific 038+ T-cell numbers or functional activit). or T cell proliferation or aethation; ard/o; (91 reduce·:, or Inhibit* Teen proliferation or activation. In one embodiment, one or more CDR regions of antibodies of the invention can be combined mcombinandy with known framework regions and CDRs to create additional. reeonsbinamly-engmeereJ, anti-CD27 antibodies ot the invention, The heavy and habit, hem variable framework regions can be denved from the same or dilterent antibody sequences. The antibody sequences can be the sequences of naturally occurring antibodies or can be consensus sequences ot several antibodies. See Kehlebmoogn e/ α>·- Pi mew Eni>iiu-?nn% 4:773 11991 h Koibingei et bl/. Protein Crymrrrire 6:971 ; 199.1? and (farter <·; ai., WO 92/22653.

Accordingly, m another embodiment, the invention provides a method lor preparing ao and-0037 antibody including: preparing an antibody including (.0 heavy chain framework regions and heavy dram CDR:-. where at least one of the heavy chain CDRs includes an amino acid sequence selected from the amino acid sequences of CDRs shown in $£Q ID NOs: 8. 9. 10.. 36, 27. 2S< 38, 39, 40. m), 5 j. 52, 62, 03, 64, 74. 75. ?6. 80. 8 Λ 88 101 105, 106; and -:2) light chain framework regions and light chain CDRs. w here ai least one of the light chain CDRs includes an ammo acid sequence selected from the amino acid sequences of CDRs shown in SHQ ID NOs; SEQ ID NO·;: 14. 15. 16. 20, 21, 22. 37. 33, .U, 44. 45. 4(5. go. 57, 58, ¢8. 69, 70. 80, 81. 82. 93. 93. 94. 98, 99. 100. i 10. ill. I 12: where the antibody retains the a In bo, to bird to CD27. The ability of the antibody to bind CD27 can be determined using standard binding assays, such as those set forth in the Examples ic..g-, an ELI8 Λ orafUSAT (t is well known In the art that antibody heavy and light chain CDR3 dmn&amp;rns play a particular;) important roh- in the binding speoii'ieity/afllniry of an antibody for an anti-ten (sec. Hail ot oi.. ./. IwimoL, 140:1665 161.3 (1992)-, PoD-menis <;t u‘.. J. Imnwuoi- ·· ......... . . . . ·· 1 . 152:5318-5329 t jv94r jabn a ai.. Innmotohit/L. 19 6-100-419 {|99gy, Kbrnka a oL. Bat, j Datum 83:252-.:.60 (7.00(0: Beiboer ,y «/.. J. Mol. BCL 290.833 849 OOOO): Rader <m. Pro,·. Nuti. Ac<hL Si i. (/.).), 95:8916-8915 t PCBy Barba-. π oi. J. Am. Chau. .37s·,, 1 lb:2 161-7162 t10,;Mi: Dtt/et et ,·ρ„ j hmnmioi.. 137.739 749 = 1996it. Accordingly, the recombinant antibodies of the invention prepared as -so forth above prei'e-ably comprise the heavy and/or light «.ham CDR3s of antibodies II 5, 1118. 3H12, 3ΛΙ0. 2C2. 209, 3H8, and IG5. The antibodies further can compose die CDR2s of antibodies I F5, i H8. 3H( 2. 3 A Hr. .2(32, 2(39, 3H8. and !G5, The nmibudies further oho comprise the CDR1 s of antibodies ; F5 JH8. 3H12, >Λ i 0, 202, 2Gv. 3H8. and ;G5, 1"he and bodice; can further comprise any combinations of the CDRs.

Accordingly, in another embodiment, the invention tinther provides and CD27 antibodies compri&amp;ing; ? .1 ; heavy chant irtsmework regions, a heavy chain CDR i re a ton. a heavy chain ODR3 region, and a heavy chain CD Re region, wherein the heavy chain CDR 3 region i, ,eiected (Vom the CDR3e of IF5. IHB, 31H2, 3A10. 2C7. 2G9. 3118. and 5G5 and i2> light chain framework regions, a Hghi chain CDR i region, a light chain CDR2 region, and a light chain CDR3 region, wherein the light chant CDR3 region is selected from theCDRos of i F 3, IHS. 3Hi2, 3 A if), 2(32. ,?.Q9, 3H8, and 1G5, w it ere in the antibody hinds (31327 The antibody may further me!ode the heavy chain (2DR'3 and/or the light chain GDR2 of antibodies !F\ i HA. 2 Hi 2. 3AI0, 2C2. 2Gv, ΛΠ8. and 1(15. The antibody may further comprise the heavy chain CDR I and/or the light chain CDR1 of antibodies 1FG 1 HR. 4112, 3A.10, 2C2. 2G9, 3H8. and IG5.

GeneAi/Inn 0Λ- /r/fiMi/fev Sequences··

In another embodiment, the variable region sequences, or portion-' thereof, of the anti CD27 antibodies of the Awe π don aie mod hied to create structurally related ami -CD27 antibodies that retain binding (/.w. H) the same epitope ,ts the unmoditied antibody? and, thing ate bt.nctionativ eouivaient Methods for identity tog residues that ears be altered without removing antigen binding are well-known in the art {sec, c.y;., Marks ei ai {PivhrbaoUtxy 11 992 s 10(7/:779-83 (monoclonal antibodies diversification by shuffling light chain variable region-·., then heavy chain variable regions with fixed CDR3 sequence eltungesb jespers ¢-/ t//.{ j(b>4? Biotechnology 12(9/:899--9()3 (selection of human antibodies from {/huge display sepertuires to a single epitope uf an umigen k Sharon cr ,//, 11986) PNAS //.94 83'8):2928-3 i tshe-dueUed mutageno'D ot' an tn\ artant amino sctd residue at the variable-diversity wsmetib one mm of an antib.-dyt; Cktsson,-/ ok {19951./. /.omtuuo/. 155(12):5947-54 u.-Viguuon m Row .md chant? e ot specified) resulting born random mutagenesis of an antibody heavy chain table region?,

Aocmdhigas. in mte aspect of the invention, the CDR 1,2. and/or 3 regions ot' the enemoered antibodies described abm,can comprise the exact amino acid sequencers'; as those of mnbodl" 1F5 His, Ml 12, '.AIR 3C2. 2GG AT IS. and IG5 disclosed herein.

However, in other aspect*- of the uwemion, the ;w;i bodies comprise denviniver from the exact CDR sequences or I F5. i Hid 3H1.1 3Λ 10. 2C2. 2Gb. 3HS, and ;G5 yet still retain the ability of ίο bind 0037 eiiecrively, Such sequettce modification;; may include one or more amino scad additions. deletions, or «οΟνΙ nations. « conservative sequence modifications as described above. Sequence mqdificabonv may also be based on die coiisonsys sequences described above for die gardenlar DORR GDR2, and. CSR3 Mqacnees of andbddiea 1135, 1H8, 3HI2. 3Λ10. 302, 3Gb. 3H8, and IG5.

Aceordioaiy, to another..embodiment, the engineered antibody may be composed of one or more CPRs that are. for esampie. %9. 055?·. 989 or 99.59 identical to one or more CDRs of amibodies 1R5. 1 H8, 31112. 3Λ U). 2C1 2GG 3BS and ICo. Ranges intermediate to rue above-recited values, ν.,ο.. CDRs that are 90-959, 05-089. or 98-10051-identical Idem it y ιο one or mom of die above sequences are also intended to be encompassed by the present invention.

In another embodiment, one or more residues of a CDR may be altered to modify bind hie. to achieve a more favored on r:.ae of binding, a more favored off-raie of binding, or both, such that an idealized binding constant is achieved, Using this strategy, an antibody having ultra high binding affinity of. for example. ΙΟ1’"' M * or more can be achieved. Affinity maturation teohmuues, well known in die art and those described herein, can be used to alter the CDR regimgs) followed by sc; corn a a of the resultant binding molecules for the desired change in binding. Accordingly as CDRt> > arc altered, changesAn binding affinity as well as immunogenicuy -can be monitored ami scored such that an antibody optimized for the best combined binding and love immunogeniciiy are achieved.

In addition to or instead of m»xi.ti!cadons within the CDRs. modifications can also be made w ithhi one or more of the Iramework regions. PR I, FR2. FR3 and FR4. of the heavy and/or the fight eham variable reports of a attdbody. so long as fhqse: modMcaddns do oof eliminate the binding affmiiy of the antibody, fGirexampfe, one or mere udn--gerntHoe amino acid residues in ihe iramexvoil regions of the heavy and/or the hghi chain variable region of a an bonny of the invention, is substituted with a genniine amino acid residue, . the corresponding amino acid residue in the human germbne sequence for the heavy or the light chain variable region, which the antibody ha·- --Iguitlcam sequence· identity vviih. for example. ;m antibody chain can be aligned to a germbne antibody chain which it shares significant sequence identity with, and the ammo acid residues which do not match between antibody framework sequence and the «ermbne chain framework can be substituted with com.\spoodln·; residues Irons the geimslnu sequence. When an armno acid differs between a antibody variable fras'nesvork region and an eqnlv alum husnati gawadme sequence variable framework region, the aniibody framework amino acid should use ally be substituted by due equivalent human germ I me sequenec amino acid if si is reasosiahiy expected that the amino· acad fails wish*η one of the following categories: (I) an amino acid residue which noncovafemly binds andean directly, id's no amino acid residue which is adjacent to a CDR region, to! an amino acid residue which otherww..· htwraeis wuh a CDR region wye,, is witlnn about d-b Λ of a CDR region as determined by computes snm'khmgn or 4} an amino acid so si sic which participates in the VL-VH interface. Residues which "ηοηνονDenny hind antigen directly'' include amino acids m posaioas itt iVamew ori. regions which base a good probability of directly interacting with amino acids ors the antigen according to established chemical tos'ces, lor example, by hydrogen bonding, van das- Waafs fos'cme hydrophobic imam coasts, and the like.

Accordingly, its one etnbi'ditnvno ast annno aud residue in the framework region of a antibody ot the invention ts snhshinted with the corresponding gessnhne amino acid .residue winch nustcovalentlv binds atUtgen uitccdy.

Residues which are ''adjacent to a CDR region'* include amino aetd residues in positions immediately adjacent to one os' mom os' the CDRs in the primary sequence of the antibody, for exasttplc. its position1.; immediately adjacent to a CDR as defined by Rabat, os'a s''DR as defined by Ototiua tvee r.%.. Chothta and Desk ./. Λ/nc Biol. 190:901 t lv&amp;7)5.

Accord 1st gly, ist one embodiment, an asnitso acid residue wtihisi the framework region of an antibody of the invention ts substituted with a corresponding gems line atmno acid residue which is adjacent to a (.'DR my ion.

Residue*·: that 'Otherwise interact with a CDR s'egion" include those that me determined by secondary structural analysis m be m a spatial orientation sufficient to affect a CDR region. Such amino acids will generally have a side chain atom within about 3 angstt'om units (Λ; of some atom 1st the CDRs and muss contain an atom that could Interact % ith the CDR arom-s according to established chermeui forces, such as those listed above. Aecosdtngly, tn one embodintests, an amino acid residue within dm framework season of an antsfi- >dy of the invention is subs? basted with (lie corresponding germ I‘me amino acid residue which whesw use internets with a CDR region.

The amino acids at sevens! positions in the framework are known to he important lor determining CDR coni smed-on : op., capable ol interacting w lib hie {'DKsl in many nutihodiex (Chelhia nod bosk. supra, Ohotkta a ai, sap fa and Tramomano vi ai. 3 Moi. Bid. 210:175 i jodi)?, all of which are incorporated heroin hy refe-eriecd. 7 here an shop' identified conserved framowini’ residues important She fTH< confonuation by analysis of She structures of seve-ai known armbodtes. The ami bodies analyzed ic'd into a limited u umbei of xSTUunral or 'Vanornrat" claw1 based on the eotdmnKuh'n ef the fDRs, Conserved frnmcwmk red dues w ahio mentbcis of a canon da; * lax- are referred m ax ' cjn>>nicur rex ;d a Οχ ( anomeat residues mclode residues 7. 75. 20. ip. 35. 1¾. r,4. 7 i.' Kb 04 and 95 of ;he he hi chum and residues 2 I, 70. 20. la. 54, 55, 7 1 and 04 of the heavy chain AddUlons! residues ·< e.. f'DR structure-deva m-mns residues! e.;u he identified accotslme A.· ihe meihodi>k/iiy oi Via run and Thodou >. iuv6 ./. Mai iUoL 2o bKOb. Nmobiy. the amine a-., ids ;a u:'.xuirup. 2. ap. o4 and ”! oi the held cham and do-eh.I and 04 -n !he heax y chain t numbering .a eecdnw to Kale a > are l now n m be capable of nue-actme v% uh the CDRs in mam .imibodtox. (he atmno ax.id4- a? position·* 35 ;n the held chant and 05 and bp3 m She heat v eit-nn ate αίχο hl-ely m buwee; w nh the CDRs, Additional residue^ x\ Inch may eSheet ( uni-οhi a:on os the CDRx can be identified acr·.seunp to the medm-dehery. ot home and \\ ink·· i iOu'2'./. Mol iuo? 274:4b d bnch residues ate termed denocb' residues and a;'e mow mstdiiex in the framewotk so woe cb'xeK nuclei ly am the , be Prune a "piaitotnV' under) She CDRs. iHe^Msesvwlitiiih:“pgfiMjpai#in the Vi.-VII inter! ace' or '‘packing tes'tduex" Inbiude ihose .resMpits at the hitfeifsce between VL and ¥B: as dofmed. for eAaotpfe., by Novotny and Haber. Pfor. Nad Acad Sri. USA. 82:4502-66 (1055· orChothia «·/ ai \upni

Occasionally, there is some ambiguity about whether a psrtknla? amino acid falls within one or mo-e of the above-mentioned categories, hr such inxianoes. aliernauxe variant end hod; ex are produced, one of which has that pa: Pee an subsiUntson, the other of which does not. Alternative variant antibodies so produced can he fexled in are- of the assays described herein tor the desired activity, and the ..preferred antibody .selected.

Additional candidates for substitution whom the framework region are amino acxix that are unusual or 'Tare" for an antibody at that position. These anti no acids cart be •Substituted with amino acids from the equivalent position of the human sermUnfc sequence dr from the equivalent positions of more typical antibodies For example, substitution may he •desirable when the ,amino acid in a framework region of the antibody tx ran: tor Unit position and the conm-pooding amino acid is· die germ line sequence is common for that position in immunoglobulin sequences; or when the amino acid in the antibody Is rare for that position and the corresponding amino acid in the germ line sequence n; also rare, relative to other sequences, it is contemplated that by replacing an unusual amino acid with an amino aetd ho ns the gemdme sequence that happens to be typical for antibodies, the antibody may he made less immunogenic.

The tents vYare". m- used herein, indicates an amino acid occurring at that position in less than about 20Cr, prelembly less than about 1()¾. more preferably less than about 5¾.. even nscre preferably less than about M3·, even more preferably ies-. than about Mr ansi even snore preferably less than about 1¾. of sequences ;n a representative sample of sequences, and the term "common", r;\ used hernia, indicates an amino acid occurring itt mure than about 25V but usually more than about 51)¾ of sequences in a representative sample, for example, all baht and heavy enatn variable region sequences are respectively grouped into "subgroups" of sequences that, are especially homologous to each other and base the same amino acids at certain critics! positions (Kabat e/ id , wquvq. When deciding whether an ammo ;« id in an ..unibody sequence is "rare” or “common” among sequencev it will often be preferable to consider only those sequences in the same subgroup as the anti bod y sequence.

In general the framework regions of antibodies are usually substantially identical and more usually, identical to the framework regions of the human germline sequences from which they were derived Of course, many of the ami no acids in the framework mg it in make little o· no direct contribution to the specificity or afftnhy of art antibody. Thus, rnanv individual conservative substitutions of framework residues can he tolerated without appreciable change of the specificity or affinity of the resulting immunoglobulin, t hus, in one embodiment the variable framework region of the antibody shares at least 85¾ sequence identity to a human get inline variable framework region sequence o; consensus of such .sequences. In another embodiment, the sari·He framework region of the antibody shares at iea.snQ0fir. 95vi,, 96¾. 97V. 98¾ or 99¾ sequence identity Us a human germllne variable framework region sequence or consensus of such sequences, in addition to simply binding 0927. an antibody may be selected to· its reteniton oi other anciVnal: properties of amihodies of the invention, such as. lor example: 111 inhibits (e.g., completely or partially blocks’ binding of CD70 to CD27 expressing ceils by at least about 70V; \2i binds to human CD.:. 7 with an equilibrium dissociation constant K.-J <«f Hw' M or less, or: alternatively. an equilibrium association con si am K.a of IU’ M'1 or greater: i'3? induces m least about 40 Q complement mediated cytotoxicity tC'DC’t of CD2? espresAfu» cells at a concentration of :0 ug/ml: and/or {4» induces, at least about -lOw· ADCC mediated specific lyAs of CD27 expressing:cetlGaf a concentration of 10 it.A ml.

ChtmuMnt&amp;iMfi Mtiib&amp;diM m LT327

Monoclonal antibodies of the invention can be characterized for binding to CD27 using a variety of known techniques. Generally, the antibodies arc initially characterized by ELISA. Briefly, microtiicr plates can be coated with purified 0327 in PBS,, and then blocked with Irrelevant prom ins such as bovine serum albumin t SSA; diluted in BBS. Dilution:· of plasma from CD27-immunized .mice are added to each well and incubated tor 1 -2 hours at 37°0. The plates are washed with PBS/Tsveen 20 and then incubated wbn a goat-unti-ltuman IsG Fc-spccitlc polyclonal reagent conjugated to alkaline phosphatase for I hour at TL'C, After washing, the plates are developed with ABTS suhsuate, and analyzed at OD of 105. P-eferabk, mice which develop die highest lifers will he used for I unions.

An ELISA assay as described above can be used to semen for antibodies and, thus, hybndomas that produce antibodies that show positive reactivity with the CCG7 immunogen. Hsbndomas that hind, preferably with high affinity, toCD27 can then !?e subcloned and further characterized One clone IVorn each hybrid oma. which matins the reactivity oi the patent cells {by ELISA;. can then be chosen tor making a ceil bank, and for antibody purification

To purify ami-CD2? ami bodies,, selected hybndoroas can be grown in roller bettfes,two-lilet splitne; masks or other culture systems. Supernatants can be filtered and concentrated before affinity chromatography with protein A-Scpkawwe ίPharmacia.

Pi wan a way. N .1 ) to purify the piotein. After halter exchange to PBS, the c-mceutratlon can be determined by ODw,using 1.43 extinction eoelitekni or preferably, by nepheiometne analysis. I.eG can be checked by «el electrophoresis and by antigen specific method.

To determine if the selected anti-ODa? monoclonal antibodies bind to unique epitopes, each antibody can be biotinylated using commercially available reagents {Pierce, Rodrioid. j.i..}. Biotinylated MAh binding can be detected with a streptas iditt labeled probe. To determine the isotype of purified antibodies, isotype LLtSAs can be performed using art reeogm/.ed techniques. I'or example, veil··: of nberomer plates ;. a π he cowed with Id pg/mi of; uni- lg overnight at T"C. After blocking wilh mb BSA, the platen ore reacted with M)

Ug/ml of monodonal antibodies or unruled isotvpe connoD, at ambient lemperuuuv for two hour·., I'he wells oar; then be rem ted wpb either IgG) or other tonvpe specific conjugated no shea. Plates are dexeloped and analyzed as described above.

To test the binding of monoclonal antibodies to hve ceils esp· easing CD2T iiovv cytometry can be used. Briefly, cell lines and/or human PBMC'·· eopri"'sing membrane' houndf'P2? ;grown under standard growth conditions) ate mixed with various οοηοοηη'ΛΟοοο id'monoclonal a nil hod lea in PBS containing 0,1 A BSA at 4 C for 1 horn Alter washing. the cells are rescu'd with HunresceindaheDd anti·· IgG antibody nnder the same conditions as th·./ primary antihod} staining. The samples can he analyzed by PACScan instrument using light and side scatter pauperises to gate on ahtgie cells and binding of the labeled antibodies ts determined An alternative assay using flu ore we-sue necroscopy may he used fin addition to or instead of· the flow cytometry assay, (ells a an be stained exactly as described above and exam mod by biuorc'sence unco-Keepy. Tb-s method allows sDual beat ton of individual cell», hut may have diminished sensitivity depending on the density of the antigen.

Amu· CDS? IgGs can he further tested tor react ts ity with the CD A antigen by Wet-tem blotting. Briefly. cel! detracts from ceils expn:asmg CD2'· can be prepared and subjected to sodiunt dudecyl sulfate polyaenslamide gel electrophoresi- After elcet;\>phu'.vs:s the -o-paraied antigens w ill be naansfeired to nl?.uxdkiSose membranes, blocked.· w I tit 20%.. mouse semnt. and probed with the monoclonal autibodies to he tested. jgCl binding can be delected using anti- laid alkaline phosphatase and developed wuh BCIP/NBT substrate tablets {Slgtua Chem, Co.. St. Louts, MOt,

Methods tor sn*tly2ma binding affiimy. cros.smn.tcti sits, and bitsdiu» kinofe» of various ;.eabGD27 antibodies include standard assays known m tlte art, for example, Biacorc’^ surface piasmon resonance iSPR} analysis using a Biacete1 νϊ 2000 SPR inairumetlf tBla-com ΛΒ, Uppsala. Sweden'. ;w described In Lx ample 2 herein. la another ernbodirnent, the antibodU* of the prvstTtf inset-don me linked to .a therapeutic motety. such as a cytm· win, a drug or a radioisotope. When conjugated to a cytotoxift, these antibody conjugates are referred to as mu\mtmoio\lrts.' A cytotoxin or OVReovii: seem includes anx agent that is detrimental t:.:· ¢¢-.,^,, hills > edK. Examples mcln#. kisoh eyiochatasin B, sranm κίιη D. ethkliitm brmmde entitle. mUonsycm, etopo-dde, K’nopo’'itk·. vincristine vinblastine. coiemcim cHxoruhcm. datmc>rubk-n. dsbvdroxx amhraein oh one. mitoxantrone. nidnnmycin, act ί man vein D. 1 - (le h y; i ί ο η· .·ο o s t e ί 0 ne. glucoccHliCokk. prove me tetracaine, bdocame. propranolol and putocnycin and analogs or homolog* thereof, Therapeutic agents include. but are not limited 10, amimetabolhes keg., me thots ovate, b-meicantopunne, bnmoguarmie, cytarablne, 5-ftuorouraetl deearbamnek alkylating agents (eap. meehiorethamhu·, tmoepa ihiorambucih nx-iphalnn. earmustiue i,BSNU; and kumr-unc tCCNUh cycioTbosphamide. bnsnUae, dihromornanmtoh sirepto/otocim mitomycin C, and evs-diehhmxh amine platinum (Hi (DOF's cisplatm), anthracyehnex ί <,<.<.. daunornhkin (Formerly daeueniyem} and doxorubicin). antibiotics te.g.. daethiomyeiix -Tonnei-lx act'monnycns t. bkmmsYcm, mnhtTunycm, and a nth ramycin ( AMOs, and ami-mitotic agents bay.. vincristine and vmhiaenneo An umibody of the present invention can lx- conjugated to a radioisotope, e.g.. radioactive iodine, to generate cytotoxto .s'adiophmmtaceuacal* for treating a dendritic -related disorder, such as an autoimmune or mRamrnaTorv disease, or grad versus host disease.

The antibody conjugate* of the invention can be used ίο modify a given biological response, and the drug moiety is not to be construed as Untiled to chasten! chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, lor example, no emyotatiealiy active losin, or active fragment thereof such as nbrin, ricin Λ, pseudomonas exmos: in, or diphtheria toxm: a protein such as tumor nee costs factor or imederon-v: or, biological response modi tiers such as, for example, iymphoklrtes, interleukin-1 DlL-i"}, interieukin-2 iMiiL-2''s. Interlcukm-b ("iL-fj"), granulocyte mnes-ophage colons stimulating (actor ("GM- CSF" t, grattutocyte colony stimulating factor ("G--CSP'>, or other growth factors.

Techniques for cemnsadng such therapdaMc moiety to antibodies aretwel! known, see. e.g., Anton et ah. "Monoclonal Antibodies For Immunoiargtiting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies .And Cancer Ί hempy, Rei-tfcid <·/td ίοϋ*Λ up. 2-15-56 (Alan R. Lass, (no. 1983): Hell Strom el ah. "AotUtodies For Drue. Delixery" in Controlled Drug Delivery (2nd Ed.h Robinson et ah tods..}, pp. 025-55- < Marcel OeUmylnc, 1987): Thorpe, "Antibody Carrier* Of Cytotoxic Agents in Cancer I'hempy: A Review", in Monoclonal Antibodies ‘84: Biological And Clinical Application--;. Rmchera et al iebv s. pp, 475*506 ( E>85p '’Analysis. Results, And Future Prospective Of The Therapeutic I,Re Of Radiol added Antibody In Cancer Therapy1', in Monoclonal Andbodies For Canee· Fkdeetion And Therapy. Baldwin, et at. levlx.K pp. 803-16 ι Academic Frees 1685.1, end Thorne efal,, "Ίhe Preparation And Cytotoxic Properties Of Amibody-Toxin Conjugates'', Immunol. Rev,, 62:116-58 068::?

In .another embodiment. the present invention pan ides a composition, c.g., a composition, containing one or a combination of monoclonal andbodies of the psesent invention, formulated together with a carrier (fsc,, a pharmaceutically acceptable earner!. Compositions eontain'uig bispecific molecules which comprise an antibody of the present invention are .also provided. In one emfxsiimerd. the corn positions inc lude a cotriblncaioiVdf multiple in. e,. mo or more! isolated antibodies of the intention Preferably, each of she Kutibonh.·:; of the coos po-or Ah hinds -o a nisimel, pre-sdeeted epitope of (bod'd

Pharmaceutical compositions of the invention also can he administered ip dombi nation therapy, ;.<·. eombmvd with other agents. To· example, the combinainM fteajsy· can include a composition of the present invention v> idt at least mie or more additional theuipeutie agents, such as ami-inflammatory agents, DMARDs ι disease-modi lying arm rheumatic dings·, immuiiosuppressise agents, and ebemmhcsapeubcs. The pharmaceutical compositions of the invention can also be admini«ierod In conjunction with radiation the-apy, •C%Fad mini strut ton whit other antibodies is also ί-ncompaswd in, The iuxeudon.

As :used: hpAfti, the terms 'WameC and 'Aliamacenficaily acceptable canied* includes any and all solvents, dispersion media coatings amibacieiinl atsd antifungal agents, isotonic and absorption delaying a gene, and the like that are physiologically compatible. Preie-abl), the carrier is suitable for irnravenom, imramw-cular. oabeutinioons. paremeral, spinal or epidermal administration in y * by injection or infusion). Depending on the route of: administiulion. the active compound, wc, antibody, bispeeific and muhl-peeifie molecule, mas be coated in a material to protect the compound To an She action of acids and other natural condition'* that mas inactivate the compound, (Examples of adjuvants which tbdy be used wifh the atiftbodies and conpiitacts ot the present invention include'. Freund's incomplete Arum, ant and Complete Adjuvant Diico Laboratories. Detroit Mich.·: Merck Adjuvant (V> (Merck and Company, hie.,

RalVvViry NJ.g AS··.: iSmbuRluic Reecham, Philadelphia. Paw aluminum salts such as liydjmide gel (atum) oralnmihaffi pbospMtep salts of edeinm, iron or zinc; an insoluble suspension of acyHtfed tyrostne" acylated sugars; erniomenlly or anionioaily derivaC-md polysuedtaridest polyphoxphaeenes; biodegradable mierospherex; cvfokmes, such a-; GM-CSF. interleukin-2. A -12. and other like factors: 3D*lvfPl.: CpG oligonucleotide; and monopbosphoryl lipid Λ. for example 1-dc-O-acylaied tnonopitoxpltoryl lipid Λ, MPL adjuvsms arc available. front Corixa evaporation (Seattle. Wash; see, lor example, U S. Pat. Nos. 4,-130..727: -1.8773-11: 4.866.034and 4.841094s. CpG-cmuaunna oligonucleotides (in y. hich the CpG chnudeoiide is unmerhykned > are well known and are desc-ibed, for example, in. WO 06/02:-41. WO 99/3348¾ nndU.S. Pm. Nos. 0,0(:8,20(1 and 3.83(062 immuitoMimulaiory DMA sequerreex are also described. for example, by Sato ei: at. Science 273:352. 199(,. P-nther ahornamt' adjasa-Us Indude, ka example, saponit-s, such ax Quit A, or derivatives thereof. including QS21 and QS7 <Aqai!a Bionharmaceubeds Inc..

Framingham Mass. r. Ext m; Digit; >mn; or Gvpsophuo or Chenopodium qmnoa ssponms: M-tmauide ISA 720 iSepp:c, Franco: SAB (Chiron. California. ( nimd States); 1SCOV18 tCM.V MF -89 (Chirorn: (he SB AS seder of aojus ants ns. a,. SBAS-2 of SBA.S-4. as ad able from Smith Klme Beeeharn, Riven sari. Belgiume Detox Uanhauzyn’M (Corisce Hamilton. Mon?.»: KC-329 tConxa. Hamilton Mont.) and other ant!noalkyl glucosuminide 4-phosplnues lAGIM: polyosymhylene ether adjuvants such as (hose described in A4') 99/52549ΛΕ synthetic uuidazuqumolioes such as imiquimod [$-2630¾. R-S37j. tlGrdspn, eb d„ Vaccine 19: 1820 1826, 2001; and res;qinmod [$-28463. R-848J (Vasdakos. e; a!. Cellular immunology 20--1: M-7-k 2000: Schiff bases of cat bon yN and amines that are consisted vdy expressed on antigen presenting cell and T-cdi suriaces, such as (acarexol (Rhodes. I el at. Nature 377; 71--71 IBOge cytokine, ehcmoklne and co-stimulatory molecules as either protein or peptide, indudsns lor example pro-inflammatory cytokines such as interferon, GM-CSF. IL-t alpha. II,-1 beta. TGP-alpha and TCP- beta. Th I nujueer$ snob as interferon gamma, 11.-2. IL-i2. 11..- if. IL--IS and IL-21, Tb2 inducers ssteh as IE-1. 11,.-3, IL-fr, U..-I0 and IL-13 and other ehemokine and co-s;imulatory gem's such as MOM* ΜΪΡ- i alpha, MtP-1 hew. RANI f;.S. TCA-3. CDS0, 0180 and CD-40L; immunoAtmuhifuin agents utreeimp ligands such ;ts CT1..A-4 ,,nd L-selectm, apoptosis stimulating protein-·. and peptides surd; as Fax; xymheLte hptd based adjuvants, such as vaofecbrt (Reyes ef .8. Vaccine iv. 3718-3780. 20()( 5 squakne. alpha-tocopherol. pdyxorbate 80. DOPG and cholesterol; endotoxin. [LPSk (Bentier. R.. Csumnl Opinion in VIictobiology 3; 23 3t). 2000>: Itgand4' that maser Toll receptors to produce Till-inducing cytokines, such as synthetic Mycobacteria? dpi)proteins, Mycobacterial protein p ISP pi.'ppdwdyww, teicholc acid anti lipid Λ: and CT wheaem toxin, subunits Λ and B i and LI theat labile enterotoxtn front E. cob. subunits A and is η he'ast ΜΚηό. protein la rods· tHSPv, and LLO tlisteriolysis? O: W(i B]/7TDv>. These and various Anther Tod-like Receptor i'TLR agonists are described for example in Katulerer ah Ado/no? Medirim , May 2007, Yol IX No 5, Λ pi ef erred immunoNtimuhnory -agent tor use in combination with an anti-CDs 7 antibods ofthe invention is a TLRD agonist, such as Poly iC A ''phamtacctttica!ts· acceptable sulf mt'ers to a salt that retains the desired biological activity rT the parent eranpound and d<ws um. imparl any andesued toxicological effects t wo :\y,. Rerge S.M, i i ai. i 077'? i, Pha.on. Set, 6b; I 19). Examples of such salts include acid addition salts and base addition «alts. .Acid addition salts ino!ads those derived frotn normwic inorganic acid?-;, such as hydrochloric, nitric, phosphoric. sulfuric, hxdrobroirhc, hxdroiodic, phosphorous and the lire, as well as irons mam owe organ a: acids such as aliphatic mono- and dicarhoxylie adds, phenyl-substituted aikanoic acids, hydrerxy aikaawc acids, aromatic acids, aliphatic and aromatic siniomc acids and the I lire. Ruse addition salts include those derived from alkaline earth meials, such as sod?urn, potassiuna* ntaamtMtnn, calcium and the like, as well as from nontax to organic amines, such as N NX dlheimylcUiylcnediarmne. N-methyialucarmoe. ehloroprocaine choline, dkuhanolwnine,, ethylenodianune, procaine and the like Λ composition of the present intention t an be .tdmunstwwl b> a \ ariety of methods Inovr;·? In the art As w ill be appreciated by the dolled wo aw, the mute and/or m:.1:.1 e of administration will vary depending upon the dCMtvJ would I he active wmpounds ertti be prepaied with carriers that 'will piotm i the compound ..w:00.0 ?ap;d u.· louse, such as a controlled release tbnnulation. including Implants. uansdemud patehiw and irueroencapstdated delivery systems. Biodegradable. biocoropaiihle polymers can be u~.-ed. such as ethylene vinyl acetate, polynnhydridfw. poly glycolic acid. collagen, potyonhoesiers, and poHDcdc acid. Many methods tor the preparation of such iorroul.rdmw ore patented os generally known to thorn skilled in the art. Vw. e.g., Suuahiiy! η?η/ ('niUioilciJ Rriaisv /May fie/;rery .Vv;,,’;u?:w J R. Robinson, ed,, Marcel Dekker, Inc., Nee York, 197s

To administer a compound of ibe invention by ecrtrnn rwues of administration, it may be necessary to coat the corn pound with, or 00 adrmmxnn the compound wain a material to prevent its inactivation, bur example, the compound may be. rdirmiisiered to a subject In an appropriate carrier, Iwr example. lipoxomes. or a dhuent. Accept able diluents inejude saline and aqueous buffet solutions. Uposomex include waior-in-oil-in-u-aterCGF' emulsions as well as conventional liposomes {Shxjjart *-i (0 i 1984)./. Nvimthuumnol. 7:27k

Carriers include sterile aqueous solutions orcl emersions and sterile powders tot the extemporaneous preparation of sterile injectable solutions or dispersion. Hie use of' such media and agents for phnrmacemu.ally active substances is known in the an. Except insofar as any conventional media or agent is incompatible with the acme compound, u-e Iheieof m the pharmaceutical compositions ol the invention is contemplated. Supplementary active compound;- can also he incorporated into the compositions

Theraneuiu. sW-mposUions typically must be sterile and stable unde:' the conditions of manufacture and storage. The composition can be ion related as a «olutiou. uucroemulsfun. liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example w ater edi.mm, polyol {foresample, glycerol, propylene glycol, and liquid·polyethylene yiycvd, and the like.), and suitable mtxnnes thereof. The proper flu id by can be rnanstauted. for example, b\ the use et a coating such as lecithin, by the maintenance- of the required particle size in the case of dntpersson and by the use of surfactants. In many cares, it will be. preierahie ίο include isotonic agents lor example, sugars, pnlyaicohols such as mannitol, sorbitol, or sodium chkuide la the composition. Prolonged absorption oi the injectable compositions can be bronchi about by including in the composition an a gem that delays absorption, for example, rnonostenrate salts and yeknln.

Sterile hijeunbu· solutions can be prepared by Incorporating the active compound in the required an scum in an appropriate solvent with one or a combination of ingredients enumerated above, as required loliovu-d h\ sterilization mictoi'fhruhou.

Generally dispersl-nts :.ue prepamd by incorporating the active compound into a -lenk' vehicle that contains a haste dispersion medium and the required othet mgred-enis from those emu Tie rated above. In the case uf sicrin· powdets for the prepataiion of sterile injectable solutions, the preferred methods of preparation arc vacuum drying and freeze-drying : honhihzulion) that yield a p;outer m' ihe active ingtedie;U plus any additional desired ingredient fo-m a previously a uau km'the red solution thereof

Oosauc rec'i;neri'· are ad;tided to prosIda the optimum debited response therapeutic response r. Fu; example, a single bolus may be administered several d;\ ;dod doses may be administered over time or do.· dose meg, be proportionally reduced o; increased as indicated by rhe exigencies oi' the therapeutic situation. For uxaiup;e, the antibodies of the

Invention may be admins srered once or twice weekly by subcutaneous or isiirarnuydular injection or once os twice sruinthiy by mux uksneot-s or intini;ui“.cuSnr injection.

It is especially advaoksgeotw to formulate parimleral composition': tn dosage unii form fos ease of ;mh'0;n;Nioukm and mfifoanhy of dosage. Dosage unis form ,v- sn-ed here so .roles s to physically dGcs-eie noils suited as unitary dosage-', for she subjects so be ceased; each ssnd corbauss a predetermhied quantity of uebo e compound calculated to prsiducc Use dess rod therapeutic elleef us asocial km with -be- iequiscJ pbars’rosocutseal easi k&amp; The \ pees fs cats r-u fos she dosage min FjcssinoI the is; tend on aye dictated by ssod directly· depessoertt oss (as she son quo chumeterksics of she active compound and the pas tic alas Usesopeusse effect so he ads ins ed. and so) die lin'hiarkms uthcrens sn she ars of cost pounding such an active eompotutU tees' she ts'earrnen? ;.sf «ensitivhy in individuals F\;smpUw of phamsaoeuiscaiiy-acceptable antioxidants include: s 1 s svasc? soluble ansirnaidssnlN such a-\ ascorbic acid, cysteine hydrochloride sodium hs'uHase. sodium mctabssulfsie, ends urn sulfite and the likes id; oil-soluble nsuiovsdantv stseh an areosTy! on imitate, butylmed hydros yacnxHe (BBAu butykued hydiOxytohserse -;BHT s. bwibsiu. propyl gallaie. alphn-tocophesof ansi she like; and i B metal oheliutnc agents, such as cstrsc add eshyiesiedusmlrse sesraaeclse acid sFOTAb sorbstol tartaric and, phoNphosi-.. acid, and the like.

For she therapeutic cmojimdbous.. fors nutations of the present intension Ihclmfethose suitable lor oral, nasal, topical is tit iuehna buccal send -mhlingnnl >. mual, vagitikl: andfoo parenteral administration The fosTnulnnous may eons emeriti;-. be prexeuieb iss unit dosages form anti mas he propa-ed by any nselhods known us the art of pharmacy. The amount of active nigreuwnf which can be combined -a. ids a css sues mate· sal u.> produce, a single dosage form will vary depending npms she subject being treated, and she particular mode of administration. The amount of acts to ings'edient which cats be combined with a carrter material to product; a single dosage form will generally be that amount of the composition which produces a tltcsapcutie cl loot. Generally out of one hundred percent, tfe amount veil! range from a bosn 0,1)01 | ter cen! to about ninety percent, of active ingredient, psvfcrabsy from stbsssst 0.005 pet cent to abour Ά) percent, most preferably from about ().(11 pet cent to about 30 per ebnf,

Fes's ms bsi'smu' of the usnsesst invention which are suitable for vaginal adrnsnlslranon also mdude pessaries, tampon's, creams, gel'·, pastes, foams or spray fosms.sj at tons consul run a such erurler'· as me known its the art to ho appropriate. Dosage forms for the topical or transderroai administration of compositions oi this mxemion include powders. sprays. ointments pastes, creams, lotion;;, gel;·;, solutions, parches and inhalants.

The active compound may be mixed under sterile conditions vs kb a ph;trn occo tic ally acceptable carnet, and with any preservatives. buffers, or propellants which may he required.

The pbrnses :!pareutemi admiBtstotion” arid "Mmlnistei'ed- f arenterally” as used herein menus modes ol administration other than enteral and topical ad minium, bom o-mully by iniechum and includes. without bumaLmn. iutravenou-'. hUr&amp;rousculsa. mm. ate rial. intrathecal, imracapiauu, uinaorhiial. m menu dim., intradermal ini raped η η teal, trau-nucbeal, suhetnaBCoos sub; uticnlar, mi ram tknbtt. sul^mp-mbt, subarachnoid. isitr,asptnstl¥ epidural and intmsteiBal iBteedoB add Infusion, •Examples of suitable aqueous and nonatjueous carriers which may be employed in the pharmaceutical compositions of the invention include Witter, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the lihei, and .suitable mixtures thereof, vegetable mis, such as ohm; oil. and htjectabie. organic esters, such a a ethyl ole ate. Proper fluidity can t'e main tamed for example, by the use of coating materials. -melt as leetfhim By the ptaimonahee: of tie: required parbde mm in the case of btspemons, and by the use of surinciants.

There eontpositions may also contain adjuvants such as prese· vattves. wetting agents. etBulspying agent- and dispersing agents. Prevention of presence oi microorganhuns may t-c onsmctl bmh by siomheutioB procedures. supra, ansi by the irtdu-htn of various antibacterial and am hue mu agents, tor example. pamben. chlorsthutaitoh phenol sorbic acid ,: ra.l the like b mm. aKe be drum able to include motorhe agents, such as .sugars, sod mm chloride, ami the like uno the compositions. In audition, ptoiongon absorption of the ntsviable plmnnacenhenl ion η tnay be doughs about by the inclusion ol agents w hich delay ub'-mptmn such m aluminum rnon-.stearate and gelatin

WteifhtTcotBfounds of the rnt-sem tiHemion are administered as pharmaceuticals, to humans and animals, they can he gh ert alette or as ,.t pharmaceutical composition containing. tor example, 0 001 to vUT ;ntore preferably, 0.005 to ?iVT. such as (1,01 to 30d > of active htgredietti in combination with a pharmaceutically aacptuble earner, idwtrdlew. of the route of adnunisSration •mlrmsed, the compounds of the present invention, which may be mvrf tn a suitable hydrated form, and/or the phttmtaccrttleal compositions of the present invention, are formulated into pharmaceutically acceptable dwmtw forms by conxentnsnal methods known to those of skill in the cut.

Actual dosage levels of the active ingredient?· in the pharmaceuricaf composiustnx oi the present invention may he varied so us m obtain an amount of the active ingredient which is eifecrisc to achieve the desired therapeutic response tor a particular patient, emripostnorn ami rood-..· ot administration, without being toxic to the patient. The .selected dosage level wilt depend upon a variety of pharmacokinetic factors including the activity of the particular compositions oi the present invention employed, or the ester, salt or amide thereof, the route of administration. the time of administration, the rate oi excretion m the particular compound being «employed, the duration uf the treatment other drugs, compounds and/or materials 'used m combination with the particular compositions employed, the age. ses. weight condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts. A physician ot reman an an having ordinary skill in the an can ;caddy determine and prescribe the etiledw amount of the pharmaceutical composition required. For example the physician or s etcdn.n am could Mat r doses of the compounds ot the invention employed in the pharmaceutical Csmtpostuon at levels lower than that required in order to achieve the desired therapeure. eiiVci and gradually, increase the dosage until Hie desired effc-ci is achieved, in geneml, a suitable daily doe of a composition of the invention will lie that amount of the compound which is the loss·, m dose effective to produce a therapeutic effect. Such an effective dose v. Hi genes alls depend up-ret the factors described above, h is preferred dam admioArraiion be intiavemrus, tniranuiscutur. Intmpei horreai, or subcutaneous, preferably administered proximal to ihe sin.· ot the target, if desired, the effective' daily dose of a therapeutic composition may he administered as two. three, tout, use, six or more Mib-doses administered separately at appropriate intervals throughout the day, optionally in unit dosage forms. While it is possible tor a compound of the present tnsemhm to be adtntmsiered alone. It is proles able to administer the compound us a pharmaceutical iosmuianon {compositions,

Thettapeuric composifipps can: he administered with reedieaJ devices known in the ait, For ©jsaniplo,:in a preferred embedtmehk a therapeutic composition of the invention can be administered with .a needle less hypodermic injection dm tee, such as the der ices disclosed in ILS. Patent Nos. 5.m.U>X N3SAS5L 5,312,335, 5.0ivUl3. 4.941,880, 4,790,824, or 4.590,550. Examples of well-known implants and modules useful in the present invention include'. U.S. Patent No. 4.4¾ 7.003, which disc loses an implamable tmero-tniusion pump for dispensing medteution at a connected rule; IhS. Parent No. 4,,4 bo, 104, which discloses a therapeutic device for administering mcdicams through the skirt; ILLS,iPstdof Me. 4,^4?,233, wihie'h discloses a medication infusion. fnmp lordelivering snedicatlon at a piecNe uUtn.ion rate; U.S. Patent No. 4,-197.3.74. which discloses a variable flow implantable infusion apparatus tor continuous drug delivery; U.S. Patent No. 4.4-39. 196. wind: dr;closer an ossr-oltc drug d·.:·livery system having multi-chamber cornparlments; aval U.S. Patent No. 4s47.SJ%, which discloses an osmotic d-ug delivery system. Many other such implants, delivery systems, and modules are known to those skilled in the art.

In certain embodiments the antibodies of the invention can be to; mulated to ensure proper distribution h> s/rw k<v example, the blood-brain harrier ί BBB excludes many highly hydrophilic compounds, la· ensure that the therapeutic compounds of the invention cross The BBB tilde bed·, they can m formulated, fm example, m upo-mmes. H>r methods of minutiae usr-ssg mu'somev hy, <. y., l .S Patent-- 1,511,111; 5. A .1,4-ΙΊ. ,uid 5,319,531. The hpo-msur s ;n«y comprise one or mom motettes vchn b ,uv selectively transported into specific cells <vvseuro thus enhance targeted drug drivers tvv, i y , V.V. Raratde 1198co J l !m /Vummue.·/. 3°: 685 rvomdasy same dug tumetiev include iolate or biotin (see, t y,, l ,S. Patent 5.410,010 ;o L ns ct ni >; mannostdes i line-mss a < / ν<1., ί 1988; Bivihem. BiotBtys. Hew Ova main. 155:1038): antibodies tP.G. Bloetrsan et ;.d. t I9v5i I'frMS Ιλί>:. 357;140: VI Ovah- a<.>/, yi99M Amimirrob. Agents Chemoiher 39;ISO); surfactant protein A rccepto; i Briscoe e? >j> i 19951 Am. ,/. Physiol 11.33; I 34 ?, difierent species of which may comprise rfar.· formulations of the inventions, as well as component* of the invented molecules; ρ110 1 Schreser >Ί αί {1994;,/ Htoi. Chem. 169:9090): see also K.

Kern a net's; M.L, Laukkinten 11994* I KBS Ieit. 346; I;10; j.j KilHon: IJ. I idler (196-3} itmtiiitwitii’Buiih 4-:.773 it; o;te entbodittient ot the invention, the therapeutic compounds of the invention are formulated in liposomes: in a more preferred embodiment* the liposomes include a targering moiety. b; a most preferred embodiment, the therapeutic compounds hr the liposomes are delivered by bolus injection to a site proximal to the tumor or infection.

The composition mutst bo fluid to the esters; that easts· ssrmgabiiiiy exists. It must be stable under the conditions of manufacture and storage and roust be preserved against the contaminating action of rmoroorsanixmx such as bacteria and fungi.

The ability of a compound to inhibit cancer can he evaluated in an animal model system predictive of efficacy is; human tumors. Ahe; natively. Shis properly of a eomposiriou can he evaluated by examining the ability of the compound n> inhibit, such inhibition 4; W/ro by assays known to the skilled praebtiones A r here peat; cal iy effective Amount of a thes'api. ttt-c compound east decrease mmo- size, or othervvsse ameliorate symptoms in a subject. One of ordinary •skill in the net would be aide m determine snob amounts based on such factors us the subject's sire she seventy of the subjeefs symptoms, and the particular cmupcfsilion or route of aumintsimoor; selected, I he composition nuts· He sterile and fluid to die re tent that dse corupoaitimi rs del tremble by sync £>..·. in addition to water, the carriet can be an isotome buffered sab no solution. ethanob poise; ; ice ex am pie, glyoercsl. propyicne gKcob and npuid polveihev lone glycol. and toe life), and suitable no at ares thereof. Proper fiat day can be maintained, tor ex an-pie. by use of coating and· as lecithin, by maintenance of neon b ed parade si/e in the case ol dispersion and by use of serf actants. in naans cases, i- is preferable to· include isotome a cents, tor example. sugars, polyalcohuis sued as mannitol or sorbitol, ansi sodnnn chloride n- the composition. Long .term absorption oi' the injectable compositions eat; be drtmgbt about, by including in the coin post turn an agent ye food delays absorption, lex example, aluminum ntonostearate or gelatin.

When the active compound is suitably pmdecled, as described above, the compound may be orally administered, for ctosnpie. with an inert dduent or an assimilable edabie carnet',. Γν. !.. scs and Methods oi the Invention

In one embodiment, die antibodies, bispecifie molecules, and compositions of the present invention can be used to treat and/or present to g., immunity against! a variety of diseases and condttlons.

One of the primary disease indications that can be treated is cancer. In particular, an antt-CD27 antibody that induces or enhances an immune response can be tn-vd n the treatment of cancer. Types of cancers include, hut am not limned to. leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, m ye lob lasts promyelocyte mytiomonescytic monocytic erythroienkemlu. chronic leukemia, chrome myelocytic tgramdoeyiko leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma. Burkin's lymphoma and marginal cone B cell lymphoma, Polycythemia vera Lymphoma. Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's macroglobulinerma. heavy chain disease, soltd tumors, sarcomas, -ark·· carcinomas, ribrrmwooma. my vtsarcoirut,! mo sarcoma, ehrnndrosareoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, e-aiotheiiosarcoma. Ivorpbairgi0sa re0mo, Ivmpbangioendotheliosareomu. sysiovloma. mesothelioma, Bxv tug's fumox. leiomyosarcoma. rhabdomyosarcoma, colon sarcoma, colorectal carcinoma, pancreatic e^|icfer,-tea§t-Oafecigiir;· ovarian cancer, prolate cancer, squamous cell carcinoma.. basal cell eaiemoma, adenocarcinoma. sweat gland carcinoma. sebaceous gland eaieinomm papillary carcinoma, papillary adenoesteinornas. CYsudenoeurcmOiTis, medullary carcinoma. btonclmgenk' cam morn a, renal cell carcinoma, hepatoma, bile duct carcinoma. choriocarcinoma, seminoma. embryonal cut chroma, WtlmS tumor, cervical cancer, murine cancer, testicular tumor. lung'carcinoma small c.eil lung carcinoma, non small cell lung carcinoma, bladder carcinoma. epithelial carcinoma, glioma, astrocytoma, medulloblastoma, cranlopharyngiorna. ependymoma, pinealoma. hemangioblasmma. acoustic neuroma, oligodendroglioma, menrmgioma. melanoma, neutoblasioma. retinoblastoma. nasopharym.geul/bdr^mcinEt'a,· esopliugeal carcinoma, basal cell earunoma. biliary tract cancer, bladder cancel. bone cancer, dram and centred new ns ο; system (CNSi cancer, cervical cancer, choriocarcinoma, colorectal cancers, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, bead and neck cancer, gastric cancer, intraepithelial neoplasm, kidney cancer, larynx cancer, liver cancer, lung cancer i smalt cell, large cell >, melanoma. ngpMblsstornm oral: cavity ca?*ce$fb£ftp, tongue, irtouth add pharynx), ovarian cancer pancreatic cancer, retinoblastoma., rhabdomyosarcoma, rectal cancer; cancer of the Respiratory system, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and cancer of the urinary system Preferred cancers include OD27-ex pros sing tomot-g,selected from the grdBp-ooiisi^f-oficbmnioil^plocydcd^ltmi^,. mande cell lymphoma, primary central nervous system" and margin's! cone B cell lymphoma,

Other disease indications for use of an auti-CDd? ami body that induces or enhances an immune response Include bacterial, fungal, viral and parasitic infectious diseases. Othet disease indications lor use ot'an ami-CD-? antibody that inhibits or seduces an Immune response Include graft reject ion allergy and autoimmune diseases. kxeniplary autoimmune diseases· mciuue, but are not limited to. mull I pit: sclerosis rheumatoid arthritis, type i diabetes, psoriasis, C-ohn’s disease and other mfiumntntorv bowel diseases such as ulcerative colitis, systemic lupus eyrhemaiosus ;S 1...1:1). autoimmune encephalomyelitis, myasthenia gravi- iMG.i, Hashimoto’s thyroiditis. Goodpasture'', ..yndmme. pemphigus, Graves disease, autoimmune hemolytic anemia, autoimmune thrombocytopenic purpura, scleroderma with ami· collagen antibodies., nbxcd connect ire tissue disease, polypyositis. pernicious anemia, id input hie Addison's disease. autoimmune associated Infertility, gkimemlouephtde, ere'comic glomeruionephruix piv>lifojun^o gkanermonepbnds. bullous pemphigoid. Siogo’n’s synch ome. psoriatic ntthdifld. msulm resismnee, uniointutune diabetes meltmis, autoimmune hcpamis. autoimmune hemophilia, aiitmromuue lymphoprolUrvaiive ' yndrc-me ; Ai .PSe autoimmune hepatitis, autoimmune iimTmphiiu. oenoimmune lynsphopsvt!ifesnative ssndrome. autoimmune eseoretmihs, Ou ill aim Bare syndrome, aris'nosck.rnsi.s and Vi.mmmerV disease Exemplary aik-rmc disorders include, 1m: are not limned io allergic conjunctE tits, 'rereal conituicttvhis., vernal ketaiooontu noli v ids, and giant papillary comunodvitis; nasal allergic disorder*. mcTadms allerstc rhmms and AnushD otk abende dwm-ess. Including enstachkut lube itching; allergic disorder, of the tippet and am,er akv-ays, including mtmom un-J extrinsic a*· Buna: allergic disorder* of die ''kin, me luduu: demi:.iTihs ee/ema and uriicana: and allergic di‘orders of Par gasirohuesdnal P an.

In another aspect, an antibody of the invention is administered tn combination vruh a vaccine, to enhenf.c the immune response against the caeca π e antigen, fot example a tnntor anitgen - to ihesebs enhance the immune iv.sp-.mse against Pa. lumort os an antigen from an mfexnous disease pathogen tto thereby enhance the immune response agamst the mfbcih-us disease patbegeisi. Accordingly in true embodiment, a saceine ant-gen -..an comprise, lor example, an antigen or ani'mem-e composition capable of eliciting an immune response against a tumor or against an infectious disease pathogen such as a * irus, a bacteria, a parasite os a fungus. The antigen or antigens -: an he. for example. pepbdes/proiems. piliysacehtdtfdes and/or ΙιρκΗ The anttgeti or antigens be derived front tamo:f8;, sndi as the various P.urmr antigens previously disclosed hereto. Alternatively, the antigen or imogens can be derived from pathogens such as viruses, •bacteria, purasnes :md/-.u tntigi, such as the various pathogen antigens previous!y disclosed herein. Additional examples of suitable pathogen antigens include, but are not limited to, the lolkiscmg:

Viral antigens or antigenic determinants can he acre, ed from, lor example.: Cytomegalovirus t especially Human, such as g.B or derivatives thereofe. t '.pstem Bear mm·-{such as sp.mde. fiavivlruses -e g. Y el leas Fever Virus, Dengue Virus, j let·, borne encephalitis ,, Iras, lap,.muse Encephalitis Virust; hepatitis virus such as hepatitis B virus pee example Hepatitis .8 Surface antigen such as the PreSl. PreS2 and S antigens desetihed tn EP-\-d-14 371: HP-A-CDO-l f'TS, and. EP-A- 1PS474). hepndus A virus, hepatitis 0 virus ,o\d hepatitis E virus; HiV i, {sucb as tat net gpL'i) orgplbO); human herpes viruses, such as gD or derivatives thereof or immediate Early protein such as 1CP27 horn HEY I m H5V2; in:man papilloma viruses; tier example HPY6, 1 !, 16. IS): Influenza virus whole five ur Inactivated virus. split influenza viros, crown in eggs nr MDCK cells, ot Yer*» cell-. or whole tie virosornes uts described b> Gluck, Vaccine, i^>2, i ts. ύ 1 6-9201 or purified or recombinant proteins thereof, such as NP ΝΑ. HA. m' M pn-feinx a ntea-ies v mux rat:rap- t Hus; parainfluenza virus' rabies virus; Respirator) S\ne>tial vims (such as F and G protein"·'j. toatvi-as (including live attenualeu viruses); smallpox vtrtn: Varicella Eexer tarns much as gpj, ji and ΪΕ03); and the H.FV viruses responsible for cervical cancer dor esample the cm1v protein-; 1:16 or F7 in fusion With a pro to: a D carrier to form Proa..·: a ΓΑ f(-> or K? fusions fro a;

1 1PV 16, or combinations -hereof; or combinations of f.6 or F.7 w ith 1.,2 ;-we Sor example VVO 96/25377),,

Bacterial antigens or imtsgemc determinants can be denied from, for example,; Bacillus spp.. including B. anShracis (eg beau h: huts to nan. Bordctdia .spp. including B. pertussis <for example pettaetm. pertussm to\m> filamenteoux hemaggkninm,.ads.n.yi;uu cyd3-1.-. fimbriae?; Borrelia spp., including B burgdorferi teg O-.pA. OspC. DbpA, DbpB), B: garinii -eg OspA. OspC, DbpA. DbpB), B. afzelii (eg OspA. OspC, DbpA.. DbpB), B. andefxonii (eg OspA. OspC. DbpA. DbpB}, R. hermstU Campylobacter spp, including. €. jeu.-ni (for example toxins. adltesitis and invasinss and C. cob; Chlamydia -;pp., including C, uuehomaUs (eg MOMP, heparin-binding proteins·, C. pneumonic (eg MOMP, heparm-bmdina proteins) 0. psittnei: Clostridium spp.. IncludingC. tetani (such as tetanus tox.tn}; C. Botulinum {for example Botulinum toxim, C. ditftdie {eg Clostridium toxins A or B);

Cor vnebacteria;n spp., including 0. dinhther-ae (eg diphtheria toxing Ehrlichia spp..

Including F. ecp.il and me agent of the Human Granulocytic Farhduosua Rieketxu spp, including R.ricketlsu; Enterococcus spp.. including E faecal is. Π. faeciusn: H.sehe· kina spp, including enterotoxie Π. cob (for example colonization factors, heat-labile tosin or derivatives thereof, or heat stable toxin), cnierohemorragie E coll, euteropaihogente Γ. cob (for example shigy tox unlike toxin); Haemophilus spp.. including H, influenzae type B (eg PRPS, norms cable H. influenzae, for example O.MF.26, high atoleeuisr weight adhesins. P.X Ρό, protein D and lipoprotein D. and fintbri.n and Ernbria derived peptides (see (or example V% .χ$4Υ46·-Π; Helieobacter spp. including 11. pylon (for example urease, catalase vacuolating toxin); Pseudomonas spp, including F. aeruginosa; Legionella spp, including L, pneumophila ; Leptospira, spp.. including L. interrogans. Listena spa., including L. rnorsocyiogencs, Mofaxella «pp including A! catarrhal is, aUo kttown as Branhamells catarrhal ss (for example high and low molecular weight adheslns and ins asms u Elorexella

Catarrhali\ (h tel tiding outer membrane xericles thereof. and OMPlOh mee to? example W007/417u\}K Mycobacterium spp . Including M. tnheruuo-hs-for example HS-Yl'tY,

Amhm·. SiSA,.-B M. bovis. M lepme. M as as05. M. puraToberculosB M smeynrum.

Neisseria spp. melydmg N gononhes and N. me;m>gmde; ri'or example capsidm polysacehuridei and conjugates thereof nunsterrin-l.unding proteins. iaetuterrin blading proteins. PHC. adhe-rinsd Neisseria rnenguiiM B tincluding <.safer mendimne vesicles thereof, and ΝχρΑ ϊ see tor example WO bCxO.C i.:t: Salmonella spp, mciudms S vypht. S. parutxphi S' sdadsnaeoulseSeermirsiidis: Shigella spp, including S M>nmd. $ dy--euteriax\ $. dextreiih Staphylococcus spp.. including S. unrests. S. epidemmiix; Streptococcus spp, deluding e. pneumonic (eg. capsular poiysaccharides mui conjugates thereof. Pm A BspA, streptolysin, cholme-bmdma pvotems and the ριντό in antigen PneumoKsin (Blochem Btophys Ac in. Ivpd.pd. 1007: Rubins et ah. Microbial fridmcrrmios. db.d.W Up, and mutant d.toxiiteb derivatives thereof'see for example WO AO/dPvri 1 : WO ·$9/03$£4ν.·Treponema *pp . including T pad alum (eg the omc; membrane proteistxt, T. deuricola. 1, hyody sentence; Vibrio spp. Including V. cholera {for example cholera Toxins; and Yersinia \pp, hududing S', entesocodticn (tor example a Yop protein). Y. perils. S', pscudotubesvuPMs.

Parasitic/Rmsal antigens or antigenic dete?mmaov east hi.· derived from, for example.: Babesia spp,. including B. microti; Candida *pp . including O. albicans-CS'vpfocoi-cas spp,, ineludingC neoiormanx; Bniurnoeba spp.. including L- hisTCsIythca:

ΡΡαποa spp,. including ;G. lambda; Leshmavtia spp.. including L. major', POsnu-dium. taciparum tMSPL AMA1. MSIW EBA, OEURP. RAPE RAPa. Serjuestrin. HliVfPi. PD3Z. L$AI. I..SA3, ST ARP. SALSA. ΡΙΈ.ΧΡΚ P025. PfsdB. PrSCfCS. FTxlh, Fled80x PfsiMO and rhetr analogues ut Plasmodium spp.'s; Pneumocystis spp.. ind.udi.ag P. xarintit Sehisoxtomn spp., including S. mansoni; Trichomonas spp., including T. vaginalis; Toxoplasma spp., including'!, gondii i tor example SAG 2, $AG3,“fg34r lYypanomxmsspp,, including!; crugL

It will be appreciated that in accordance vault This aspect of the present invention antigens and antigenic determinants can, be used is; many different forms. For example, antigens or antigenic determinants can be present as isolated protein;·; or peptides {for example in so -called "subunit vaccines’! or, for example, as cell-associated or vlrus-asvociated antigens or antigenic determinants (tor example, m either live or killed pathogen attains». Live pathogens will preferably be attenuated in known manner. Alternatively, anugen.s or untigentc determinants may be generated in situ in the subject by use of a polynucleotide coding Γογ an antigen or antigenic determinaptiMin· ao^dal - vaccination"), although it wilt be appreciated that the polynucleotides which can be used; mih this approach are not limited to DMA. and mas- also me bole RNA and modified polynacieoidea as dtscnased ibov'e,. in «m* embodiment, a vaccine antigen can &amp;l»o be targeted, for example t&amp; pan.wukk cell types or to particular tissues. For example, the vaccine antigen can be targeted to Antigen Presenting Ceik t APCst, tor «ns ample by tt.se of agents such as ani.tbod.kes targeted to .MT'-surface receptors such as DEC -fOA for example as discussed in WQ 2(s0v/0619% iCcHdes TIterapenftes, Inc?, or the Mannose Receptor tCD206'> lor example as dket.tssed tn WO OaO-IO i GO ?Medarex. Inc. t.

For use in therapy, the antibodies of die I use· tit or? can be administered ίο a subsect directly i? x, in νόο}. eithei alotte or with othet therapies such as an mmiunostimulatory agent, a save use. chemotherapy or radiation therapy. In all case··, the KstdoodUvs. bis pec dies, compositions, and immunoshmulaior} agents and other therapies :w adminixteted in an effect iso amount lo exert their de«ired thcrapf.sk ;c eiux.t Thu fern! "effective amount" refers totn.n .unouui noee-sarx or suificient t>i ieah/.e a destted iw-F'gn effect. For example an ei'IKihe amount could he that amount iieee-'s.tr\ m elirntn tt>.· a tumor,cancer, or bacteria! sioti or t\maal intocd-m. The ΗοχιΙγ·· amount ί<υ any pat titular application can s ary depending on such men vs a - the disease or condiUvtt bcina treated, the particular antibody being ad mi m-A: red. the si/·,· ol the subject, vk the seventy of the disease or condition One of ordinary Apt in the art can stupinerdh determine the eiteethe amount of a particular molecule without necessitating undue experimentation

Ereterred routes, of admimstmtion..for %act tnes uts'huk, for example, injection w:.e., subcutaneous. imraxcnous pasetuetai. aktkpemonetU, mirumecal; 1 he ;n;e·. uon c.m be in a bolus or a continuous infusion Other tomes ot administration include ora! admit pktradon.

Antibodies and btspcnllc molecules of iltc invention also can be coadministered with adjixaikK and other therapeutic ageiks, h v.nil be appreciated that the teem Woadmlnktrred'' as used herein includes any or all of simultaneous, separate, or sequential administration of the antibodies and conjugates of the present in', moon with adjuvants and other agents, including admiuktrattou as part of adoring remnant The antibodies am typically· forniuildted in a canker alone or hi: do.tPMhhtt©ft'--wifli. sacji sgetfts. Examples id such earners include solutions, solvents. dispersion ntcdia. delay agents.

The .u'se of suc h media for pharmaceutically active substances is well known In the art Any other conventions! earner suitable for use with the molecules falls within Ua- -.cepe oi the imituU un'cntion,

SuUabie agents foe bo-administration with the antibodies, conjugates, hi specific··., and eompo- inen·-· Include ether antibodies, eytotoxin-. and/or drugs, as well, as udjuwuUs, mtniuitosdmulalory agents and/or immuniwuppawrlvc' agents In one embodiment. the agent is .1 chemotherapeutic agent. Use antibodies, bhpeclfics. and compositions can be administered in vommuation with radiation. rswmoihentpeoik agent’- suitable tor coadminl strut ion sslth the antibodies and conjugate- of the pre-ana invention tn the treatment of tumors incicde. for example; 10x0]. esmchalaxm B. erumteidln D, ethudiunt bromide, cinetrne. mitomycin. etoposide, teuoposidv. vim n-.tme χ inblasime. t.i'ichteiu, doxorubicin, datmorubiesm dihydroxy unthraein dionv. rn to x a a none mirhuntnetn. aciuuanyem P. I ..1 e h y -d ro t e s 10 \ t e eo n e. auscocorpemdx. procaine, tetracaine, hdoc.tmo. propmnoloi. and nmomycin and analogs or hour docs thereof. Further ugems include, for example. «niimctabolUes one.. ;nethoircx.ae boneivaptopurine, b-thsotmamno cyinrisbine. vflnoroumeii dvcarbafme). aikyiating agents tc.gc. meehluretiiamlne. ihioena ehiorasnbueil, rnelphalan, earminitioe (BSNU s and iornnstine K’C'Nl!). cyelotttosphamidc·. hnsulian, dsbromonunmioL streptoxotocim raitomydn <11, and cis dichioivdtanune platinum (IB iDDP) cixpiannh amhracvcimes ie.y., daunornbivin tYorrnerh daunomyems and do\Di ubtcln). antibiotics tc.g,, dtictutomyoin {formerly aetinomyein5, bleomycin. rruthmfryi m. and nnthmmYcin t AMQ). and ami-mitotic agents (r.g „ vincristine and vinblastine) and temoxolomidc.

Aaenis that delete or inhibit;mnιua0xappres1 ve .activities, for example, by immune ceils ifor example regulatory T-ceils, MKT ceils, macrophages. m>clold-derived suppressor cells, immature or snppresstve dendritic cells) or «oppressive factors produced by the tumor or host cell·'. ;.n tiic local mkroenv irornnem of the tumortfor example. TGfbeta, indolentnine 2.3 dioxygenase IPOs. may also be administered with the antibodies and coaiugates of the present invention. Such agents include antibodies and small molecule drugs such as IDO inhibitors such m- \ methyl tryptophan or derivatives.

Suitable attents I or coudniiai-ttrytioa with the antibodies and fu-yx-'clilcs of die preyeaf hAxstMow ifor treafotent of: such trnmnae disorders: include for oxampie. immunosuppressive' agents such as rapnmycin. cyclosporin and fKSQb; anti-TNFa agents such as etunercept ubnlimumab mid inllix'nnnln and steroids, Examples oi specific natural :xnd synthetic xmroids indude. «or example: aldce-acrone, dcvlomoiiwxoae. bedroeth-tson'^ budesunide, vioprednol, eo'drsone, awn vaxof Jeoxyeoaone desnnide, dexo\ynensXi'nt\ dexan-erhasoee. dilmosoeoaolone, ducOroOntx fiinncahasone, ik<msohdr-, flnoernoione. i'luodnonide. lluoeomn butyl, il no record sene, iluoroconolone, Iknxror.nedknone, fluranUftmolone, fluticasone. hufemonide, hydrocortisone. icomeiha&amp;one, mepfednixone. methyd prednisolone, pararoeihasone, prednisolostkwpradmsomn t-xoeortol and triarndnolonec Suitable agents for coadroioi.stratiort with the antibodies and bispecifea of the present invention lor inducement or enhanecsoetn of an immune response indude, for example, adjuvants and/or irorounostiroulatory agents, non·limiting examples of svhieh have beers disclosed hereinbefore. A preferred troutum>stinlXi!atory agent is a TLRo agonist, such as Poly 10.

The present insemion ο further illustrated by tire following example4· which should not be construed as Oat her dmruaa, Ί be contents of Sequence lasting, figures and all relcvences. patents ansi published parent applications died duougheuf bus application are ex press k incorporated herein by reference.

Gemmalhm of €I>27-Speeifte Human Monoclonal Antibodies \ ha rum anb-CD27 monoclonal antibodies were generated by immunising die HC2/KCo7 strain of HuMAb%) transgenic mice Γ'Hu MAh" is a 1 rade Mark of Medarev.

Inc., Princeton, New Jersey! with a soluble human CD27 antigen. HC2/KCo7 BuMAb mice 'Άηre generates! an described in U,S, Pat, Non. 3,7 7(5,479 and .AaAASfX;, the entire disclosures of which are hereby incorporated by irteuence

Antigen iaid hum υ me at ion. The antigen was a voluble fusion protein comprising a CD27 extracellular domain fused with an antibody Fc domain (recombinant human CD77-Fc chimeric protein ;RAD Systems). The unrigen was mixed with Complete:· Freund's (Sigma) adjuvant for the first mimnni/.ation. Thereafter, the antigen eyas mixed with Incomplete Pram id's (Sigma)* Additional mice we?e imm united oath the soluble CD27 protein m RIB I MPL plus TDM adjuvant system (Sigma) 3-2.Ί ntierogroms soluble recombinant CD’27 antigen in PBS or 5 x I i/‘ CHO ceils transfected for sail ace expression of human CD7.7 in PBS vve-e mixed 1. i with the adjuvant. Mice were injected with HH? microUtet's of die prepared end gen into the peritoneal cavity every 14 days. Animats that developed antbCD27 titers were given an iv infection o( 10 mictcarams soluble meombmam CD27 tm been three to four days poor to fusion Mouse spleens were ha; vested and the ; so la fed xplenoeytes used forhybridoma pimuimbom

Hybndoma Preparation: The P3xf>3Aa8.0b) murine myeloma cell line (ATCC CRL 1580) was used for the donnas. RPMI 16-40 (hnitrogen) con da mag. I0vc FBS, and was used to culture the myeloma cells. Additional media supplements were added to the Hybridcma growth media, which included: 3vr Origen-Hybridoma Cloning fmetw i Igsn). I OB FBS (Sigma). L-glutamine (Gibco) 0.1 vi gcntarnycin 'Gibcou 2 mercnpKfcthauot (Gibco). HAT (Sismut i.Ox Iff4 M hypoxamhiue, TO x IQ"' M amiuoptmin, 1.6 x 10' M thymidine), or IT! ; Sigma; 1.0 x 10’ M hypos an fb in a. 1.6 x 10' M fhvmmmm media.

Spleen eelia were mixed with the Pdx 0 3 Ay S .653 η we lorn a cells in a ml ratio and pelleted by cent rifa gat ion. Polyethylene glycol was added dropwtst with careful mixing ίο facilitate fusion. HyFrinomas were allowed to grow our for one to two week', until v imt-le colonics become established. .Supernatant was harvested ami used for initial screening lot human !y(S via FI.ISA using a human kappa chain specific capture and a unman Fo specific

detection. Tg(3 p^.--hi-/e supernatants we-e diets assaved for CD-7 spceidcnx Go fiuvr cytometry or u\mg a ELISA to detect ann-CTCC

Hybndomss producing specrHe human monoclonal antibodies s human mAhs; IgCA 'acre xubdoned and expanded. The human m Abs produced xxere thee, purified b\ protein A'column chromatograph) aci..ordiag to s-andatd conditions vxhich led to the isolation of a number ol antibodies' ot patttcular interest which \x ere designated ms IR7-! Be tabo referred to hetvm as 4B7h 3HI3- ICS ia!w referred to hetdu a<· 4HI2 t 1 Eh-1 VS5 toNo ret erred to herem as tide 2C2 1 AiO «also referred to heiein as 2C 2 ’ - f:Cd- if);! taKo referred to herein as 20 ;B . IHS-B4 tulso referred to herein ;e-- 114¾) , 0112-1 R12 >n';-<o referred to her-;·;n as 3Π 12;, .4( ltd Λ i 1 SaNo referred to ho;errs c\s 3G1 t (1S IOC 4.A2 -B! i • also refer trU to hereto as 4 Λ; . 3A Hj-G i 0 u'd'-o re?erred to herein aa 3Λ KO . 2(i! I · Bn taJso •/ferred to herein as XI i ;p 41 1 i ; -G 11 prEo referred m herein or 4HI 1 '> 2H3 41.8 toNo referred to he rout as 2; S3), 4Λ7 B3 tabo adened to herein as 4 Ah )418--; H S ; ah-o referred to herein os 3H8 -and IG5- 189 mbo refen ed to herein as ΚΊ5;.

Tfee hy;ipidptms were aJ so sctepKed for A i-tff rhesvis macspxte GD27 and dll were positive Bar Binding.

Example 2

Determination of AftMly and Rate Constants of Human mAbs l>y Surface Plmemon Resonaoee (SPRl

Binding affinity and binding kinetic-; ot' carious human untECD27 antibodies", front Example I ere re examined bv 8 i score!surface plawnon resonance (APR; analysis using a Biacore!'? 2000 SPR instrument tBtaeore A 8. Uppsako Seeders > according to the manufacturer's guidelines.

Pun tied -eeorablnarh husrutr; C027n'NFRSF’XFe chimera iJXv.D Systems Catalog No. 382-CDt protein was covalently linked to a BiaeonG v! CMS sensor chip tcurhoxymethyiaied dea-ran cm chess By attached to a gold surface; Rtacore Product No BIG 1000-141 using standard amine coupling chenibt-v with an Amine Coupling Kit provided by Biacore according to the manufacture-'s guidelines >BiAcore Product No. PdG-IOOOoCh comprising coupling reagents ty hjdroxysoccinunirie N; IS ° and I -Etbyl-o-i.C dimei.hy;a.oiinopropyl) eadaodiltnide hydrochloride (EDCn, Lire. lex-els ot ligand were immobilised to limit any et'teus ot mass transport of analyte on kinetic parameters, such that: the Κολ.χ observed was In the order ot' 100-41)0 RC.

Binding wax measured by flowing the antibodies aver the 'tensor chip in FISS-HP buffer (HBS-EF buffer, Biacore Product No. BPwOOI-88: 4-O-hydrox.yethyF»-! · p ρera?.ir;cutheficsi ifοηic acid < HBPHriri 0,01 M, sod sum chionde 0. 15M eihyicnediamineidfaaccUc acid (EDTA) VoM. Surfactant F.?.0 0.0()5¾}. at eoneeah<mo;u tanging from 1.06 to 50 nM and ai a How rate of 30 uuYmmae for I HO seconds, The antigen· antihods association and dissociation kinetics were followed tor enher -130 seconds or i200 seconds for antibodies waft slower dissociation rates.

Corresponding controls ssere conducted in eai.h case using a blank mm. eu H with no protein immobilized io.r '"background" scriou; union. Sequential iniectiotis of 10 mM lid for 1,0; seconds at 50 μΙΑηΙη followed fey tflmM Glyeirie p|I 2,0 for ;30 seconds af 30 μΐ/rniu was used as the regeneration conditions throughout the study.

RiaeorcT BlAesatnaiion software session 3,2 (Biaeore ΛΒ, Uppsala. Sweden) was used in. each case to dense kinetic parameters from the concentration series of anals te dffistcd in HBS-EP running buffer Hie association and dissociation curses were fitted to a 1;;1 OLatuunuir binding model using Biacore! vl BlAevaluahou softv.-am (Biacme ABr acoorsling to the nramifacutter's guideline'

The affinity and kinoov pmutoetos gwuh background subtracted) as determined are shown in Figure 1,

For each antibody, the figures shown use the mean of two separate series of experiments, using separately prepared trim cods in each case, (where ka - sate con star U oi association, kii ~ rate constant of dissociation, Kr: ~ dissociation equilibriuifr<toft$ta»£ (measure of affinity), Κλ- as*otoatH'n ouuilifet sum constant, Rmax - maximum SFE response signal h

Esan title 3 ELISA Assay to Determine Human mAh Binding Characteristics t>n CD22

Tlic;other plates were coated with soluble or recombinant human or macaque 0,)21 in PBS and then blocked with 5T- bovine serum albumin in PBS. Protein A purified human mAbs and an isotxpe control xxere added at saturating concentrations and incubated at 7>T€. The plates were wa-ihed with FBS/Tween and then incubated with a ymmanddriimatt IgG fc-specific polyclonal mag cut conjugated trs alkahuc phosphatase ;u 3'7°C. After washing, the plates were developed with pNPP substrate f 1 rng/oti y and ana Is zed at OD 403-650 using a microtuer plate mader. Representatives binding curecs tue shown ur Figure 2.

The Jesuits were afro used: to estimate tie· 50% SatMaizng coneerftradoo |C ygtejp.t».·^· parameter fit curve) as shown in Table 1 bekm.·.

To establish that eynomolgus mttequea arc a «Levant model for testing ami-ODT? roAbs, varmus conceutraimns of purified mat aqua CD.?.? or human CD 2? were captured to ELISA plate-. with ami-Flag antibmiy. iollowed b> incubation with anti human CD'2? mAh; A goat amt-human hjG Ee HEP ant?body and substrate doper Blue TMR awe used for deteebon. Result" are shown in Table !.. v,inch indicate --0011.:0 binding to CD27 Irorn maepue and human. Representative binding curves for antibody HF5 are also shown in Figure e. W*tl

Oftaracferitraftort of selected anii~CD2? mAh

in a further experiment, to establish simitar distribution of iF5 binding to peripheral blood cells. PBMCs were isolated from whole blood or 5 .humans and > eynontoigu* macaques. The eel Is were stained wnh IP 5 mAh together with markers to delineate the major Tceti and B cell populations that espies;- CD27. The follow my, table ("Table 2) summarizes the mean 0 standard earn of results lor human nod macaque cells with respect to the percent of cells expressing CD2? and the intensity of expression (MR). These data establish a similar distribution of ΓΓ5 hiudiu» to peripheral blood cells from human and mortkey. mki

4A: Blocking of $€070 Binding by ELISA

Til·;· effect of r.he human utAbs from Example { on the binding of soluble CD70 ^.(7.)70} to (7.)77 protein was measured by ELISA A noerotuer piste was coated with i upon; soluble recombinant hut nan C(.427/Fc chimera from R.&amp;D Systems at luamvX.., then blocked with ?Q PBA The anti-CI )27 antibodies S' [final] = 25gg/mt.i were premked with «oiubU: human recombinant CD7()~biotin from US Biotogicals t[ttnal] L.Pita/nd .; and added tsa the plate. CD27-captured r<7D70 was detected with sueptavidm-HRF and substrate Super Blue ΤΛ1Β. The -exults (shown its Ά Mocking) are shown in Figure 4 with controls as indicated These results show that several of the antibodies (including 1F7, IHS. 3i t I 3 and ; A4 ) had the property of blocking or at least MgniUcarttly Inhibiting die binding of sCT470, 4B: CD27 mAh Binds to 0427 mi Ifimiait Lt mpltoblayoid Cell Linos aod Blocks Ligand (CDTOt Binding

Binding uf umi-itumnn €D27 mAh ITS to human Lmphohlastoid e-.-ll lenes, and blocking of s€D ?n binding. was analysed l>y How eyn wr-ny using a Bet.ton Dickinson F.ACSC'amo It flow cytometer The m-nit-. are shown In I ignm 5 and show that IF^ effeciisely bind-; to a να riot} oi celt linos mid oumivdmoh inhibits binding >4 sOD7d. 81 oil lug of Human toAbs to Cells Expressing Human CD 27

The ability of anti-CD27 human rn.Abs to bind to 0027 on cells espies sing human OD27 on their surface was Investigated by flow cytometry as follows.

Antibodies were tested for binding to human cell sines expressing human P2 ' on then Burtace. therein A purified human mAb.x 2C2 )H3 and IP5 we··?: incubated with juikut, Rail. Ramus and Dnudl ceils expressing human (71437. as well as control eel is at 4X7 Alt antibodies were used at saturating concentrations. After i hour, the cells were washed with PBS containing 0.1D BSA and 0.03*7 NuN; (PBA t and die hottnd antibodies v>erc detected by mumming the celts %vhh a PH labeled goat untt human sgC Fc- speclfic probe, at -L'C, The excess probe was washed from the cells with PBA and the cell associated fluorescence was determined by analysis us in a a 1.,81¾.1 νϊ instiument fBD Bioxclenees. NJ, USA} according to the manufacturer's directions.

As shew n in f'igcnex (> and 7. the human m.\bx demonstrated high ίονοΐ binding to cells ex pres sing human CD2 ? 'these data ueomnsirnte that these ami bodies bind etlieiemiy and specifically ta· human (1.)27 expressed ;.·η live coi 1 o compared !<> die control cel·?, mmmji

Croas»b1oekϊn«/conipdtill on of Human mAhs IMer mined by Cl. ISA Λ microritet plate was coated with it recombinant human CD27-Fe chimeric fusion protein. then hktoked with 5Ά BSA in PBS. Uncrmjugated human mAbs <20 pg/cnLi were mixed wnh horseradish peroxidase labeled secondary antibodies (0,5 pg/rni..}, then added to Ote plate and incubated at 3?X1 The plates were washed with PBS/Tvreen and developed w ith TMB substrate and analysed at OD 450 irons a roierother plate reader. The results. shown in Insurer 8 to 10, indicate that a first sei of the human rnAbs (comprising mAbs IΓ5, i H8 and 31117) cross-competed with each other {see f igure he that a further set of the human rnAbs icomprisms mAb« 2C2. 3118. JOS. and 200) aBo cross-cornpeted with each other (see Figure v'h ami that human rnAb 3Λ 10 binds a unique epttope but may bind at a site distinct from but possibly close to the binding sites of m.Abs IP5. 11 IS. and 31117 since these antibodies were able to pmi tally cross-block the binding of 3Λ10 to CD27 (see Figure H)r C®i«|llhmeixt fk’pemkmi Cellular Oykrtoxidty (CUCC or CD(')

Targe; cell·'-;! \mphmun Ran cells} sxere culm red (in AIM V medium'· for 1 -2 boors at 37Ύ.’. 5'eCO^ in the presence of anti-CP27 amibodics and rabbit complement (final dilution of !; 15) in a tlnal volume of jpOnl. Appropriate controls with a lens signal (targets only} and MAX signal it argots with Ly-ei*'5 detergent at l2v< fot a final eonccmmimn of 4vf} were included as well. Ceils were adjustedm I :s iO'Vrnt aud 50ul were added to each wed ito give 50.000 eelK/wdr? Wells were then resuspended and lOOul ot the edl gaypensjou was transfenecl to an opaque, white plate To each of these wells. lOOnl, of Ikomegu Cell Titer GIo reagent was added and the plate, was mixed ten' 2 minutes at room temperature. The plate wss allowed to incubate for 10 minutes to stabilise the luminescent signal I,·. cranes*, enee was recorded on a Perkin Elmer Victor X4 plate reader. Cytotoxicity was deon mined with the following forrnnln; ; i00-((sampk MAX ;7ileak MAX))» \ 0)0.

Results (show n -is w lysis) are shown in Figure 11 from which a c-ab'lfe· ^€ϊι·'^δί4·^«.6!0:Γ# the ants -€D2? antibodies displayed significant CDCC activity.

In a t'other experiment, target cells (Ramos cells» were washed-ami loaded with Calcuo AM (Molecular Probes) Loadedceils wc-t tin:·: washed again and resuspended to 1 x lO'Vml in calm re media dRPMI w 10¾ PBVh Target cells 'Acre cultured for 2 hours el MX\ 5w€G.> in the presence oi a«tt-CD27 antibodies and rabbit complement Ulnal dilution, of 1:1*») in ;.t tins) volume of I50uh Appropriate com ml·; with a leak signal (targets only) and MAX signal lungers with Tnton X-100 at 20w for a final concentration of 2¾) were included as well. Following the incubtuiory 75ui of supernatant from the wells were· transferred into ass opaque, black plate. Fluorescence (Ex 485: Em 5351 wats recorded on a Perkin Elmer Victor X4 plate reader .Specific cytotoxicity was determined using the following forsmda: le s peri sum o'a I ~ spontaneous lysisV'imsximum lysis - .spontaneous lysis; s 100. The. > exults. shown m Figure 12, indicate that ami CD27 mAh (Mm) showed at least 10¾ CDC activity In Ramos cells m ass antibody concentration of yusdrd.

Arsis body Dependent C'eH-nsedioied Cytotoxicity (ADCC) 'Targets eeiu -Lymphoma Kuji cads) were washed and loaded vmh BA I DA reagesu -reiki·: Llnicn. I.oaded cells were then wad ted again and resuspended to '.\ 107ml in culture media <RPMI 10¾ FBSr. Efircto< cells were ptepared and adjusted to .mpsopruttc c<mcen?raii<;;w lu vulture media to yield desired eflVeioi'target ratios · tUO:1 -'Ήη 1». 1st a round bottom phste. largn cells. effector cells and antibody wese added in a final volume os 150t?l. Appro;;: iaie controls %vew used, including a leak signal - targets only r a spontaneous U-sw Canal (targets ·+ electors'), and a maximum lysis si gnat ίtargets ; LysoF detergent a? ; ?/( fora mad concentration oi'47ek Cells were pelleted ;n the plate and incubated for 2 bouts at ?7°0, 5%CQ>. Follow hug the incubation, 2linl of supernatant ftont the wells were transferred into an opaque, white plate. To each of those we IE. 200ul of Europium solution«Perkin Elmert was added and the plate was mixed lor l.i minutes. Tune-' ntsoKed fluorescence was recorded on a Perkin Elmer Victor X4 plate reads'?. Specific cyti.uos.ldty was determines! using the following JormtsUt: (experimental - spontaneous ly;sis|/(max Imam, lysis - spontaneous lysis) s M)0. Results jybowm aslysis) am shewa in Figure 13 from which h can be seen that a number of the anti -CD27 antibodies displayed significant ADCC acid ity.

In a fnrher expeiimeui. laiget cells (Lsmpborua Rauvis and Da$£li sca-otcd and loaded 'a uh Cakvni AM i Mohxiuar Pn>ηη··; 1 widen cells we*e then washed again and resuspended to lx IG'/mi in milium media < REMl + I OR- FBS) Effector ceils were {.at* pared as id adjusted m appropriate r ora emr.hlons in eulmw media to > ie id desired effector, target η daw i 7 3; 1 s. In a round tmitom plate, ice get cells, effector eel Lx and am I hod) a are added in a dnai solum c of ] 50m. Appropriate controls wore used including a leak signal Darya.· lx on!);, a spontaneous Asm signal uaigeA i effeehns), and a maximum lysis signal uargets t Triton X- HW at 20A· |dr a final soneern ration of XV i. Cells were pelleted hi the plate and incubated tor 4 houo' at 37' 0. F'hCO,. following the incubation, 75a 1 of supernatant from the wells were trims [enitsdlnto art opaque, black plate Flu-'rescent e {Fa 485: F.m 535 ί was recorded on a Perkin Elmer Victor \-j plate reader, Spcwifw cxmm-shmv was determined using rise following formula: tvspeiu'uemd spontaneous hsixs/i maximum Iv4\ -- sponuineons Iyxio) x 100 The results, shown in Figure 14 indicate that amF-OFX!? mob t iffo showed at least !0'V ADCC activitx mom ~;u red a-, speedtc cytotoxicity* in Dandi and Kamos cell··: at an antibody concentration of dfig/ml and ratio of efiector: target mlh of 7mh

Antibody Sequencing

As described above in Example ), human mAbs front hyp: Idwmw producin'* specific human mAhs tg.G were perilled by protein A column chtornaMgraplu xshieh led to the isolation of a pane I of antibodies * human mAbx > of partieuias micros!, i he V;; and Vh coduig regions of human m Ahx 487, 3Η i 2, i FF, 2CA 2GS. IH8, did I d. 3G1 I I B1 0 i -FA3. d.-AK), 2G1 I, 4FH I. 2143, 4Λ7, 3H8 and IGF were identified «slug BAA from the corresponding hybndwnas, RNA was reverse transcribed to cON \, the V cooing regions were amphued by PCR and the PCS product was sup mowed The iollowing e the nucleic and amino acid sememe ex of tin* VT; mui Vh regions of the human mAbs nn the care of the ammo acid sequences, the Complementarity Determining Regions iCDRx; are aniledined;. 3HSVIIlVHV7Q)7.27ij;Bli

Vu nucleic acid sequence fSEQ ID NO: F) luyeaauppygcmrmiitaeettncenpnpeainitaposgpipiccriOtpigappipcapctgetppaeictggasgacgotm. rnccagectssspiY^iccctsapocicti. cicfgcos'ccicta'gaucaecntspmpnstmyatogccippgiccpecagyctcoa ««e3a;i-?^cii?£a^Mff£cigi)gL\iataiauaecaaaatp£a;^'fiLias:aaaiaci;itai2avK'ictsigaag«scc«:iticiicc3i!; icca<?;ti;.'if aaegucaagiuiaencisueuacaaaig^acagecieaeaguegaggacauaeiagigianacigigigagasaa c\gg^gaiojKKnggiactiv'Saicu;tssKi'vra;oacaci.cig«u:;ii.iaK'k:i:u;;!

Vn untmo acid sequence (SEQ ID SO: Os (im lading \;gnnl peptide in underlined I lan e a);

3//- / (?/..VU Α-Ϊ 7. V, \//,/·ϊ i VOL EVQ1. V ESGOGL V DPGOSLR ESCA A SGFf FSSY W M AW V R OAK^KOL£\\LOSiKfixiSr.KYVVDSVKORriLSRDNAKN\SLYL<iMNSU^E.DTAVY

V^yHEUiMDwY H3LWGR0 Π-VTVSS V:; "maun'e'" annno acid aequenee (SEQ ID NO; 1) excluding xjgual peptide: EVOl. V ESGOG l· VOPGT.SLR t.^C A ASGPTESS Y W MAW V ROAPGKGLE W LON IΚΟΓΧ Ϊ SUKYVVDSVKORFilSBDNAKNSLVLQMNYSi.RAHDTAVYYCVRjMM&amp;mi'DLW^

RGTLVTYSS

VaCDRi (SEQID NO: U GRASSY W Vu CDR2 iSF.Q ID NO; v): (Ni>D£;SEK

YiiCDR.5 f$EQ ID NO: Kin V R\\\ .(1 Nil)WYFINE BUB VK#2|VK 3-f R3K.fi

Vj mReie acid \eqncnee (SEQ ID NO; 11)

At ggungceccagclengt. ncRiccsceRe· nefetagetcceagafacena a naaaaun ingug:«.nengu.tecagt. cae ceiu|cudne:ci.agsggaaagagecaccciaiceigcagaaecsg?eagagigPin.R.agean UuemggRa aacaxxsaae c;ppivaaacic-;i,aaae?rcK'anaafpaiacan.caai.aaaai.aaamgoa:c-;i:agt:c.4mUcagiagc:!.popa!c;qgt;a caa;:Kn:cac:c;caicatiaaaaaC£ia2ase<;aaaa£a:tupoaprnaiiactplaapcapo:Maaaa;;i.aa:nx:na;aacauc!; acaaaaqaacc.avaataaaa.iii.aaa

Vi.nmlra.i acid ai-xpatau.a (SEQ ID NO: 12) undudins: signal peptide in underlined nahcG: \it; \ΡΛθυ i' /./ /./ njjsrmosvu qspat lsls pc; p:r atlscr aspsydsye aw yqq

kKlOAPinjJYDASNRATCORARFSGSGSGlTd-rLllSNLEPEDFAVYYiOORSNWPF

TFGQGTKVFJK V-. amino acid sequence iSEO ID NO; 13) excluding uigna; peptide;

ElV LTQSPATLSI .SPG ERA' i'LSCR ASQS V DSY LAWYQQKPGQAPRLtiYDASN R ATG I PAEPSGSGSGT DFTLT1S N1 .EYED FA V Y VtiQQBSNWPFIIAlQiTrEVEIR

V, GDR1 (SEQ II') NO: M): pSYlRY

V; CDR2 (SEQ ID NO: Er DAS V, CDR3 Vi HQ ID NO: lOi;OORSNWVrt: A ilirther light cMsb was 8|ss :fodtd to he Mtlve as thllows (though: only the above light chain <3Hb- IBM VK 22) was toed in ;ho atsove Examples; V;. uncled aeid sequence |$E0 ID NO: I?)

AlpguasccceasctcagtttctcUcckxigaacfctggcicccagafaccaeegsagaaatigtgugaeacagtetceagccae cctgtcttt?icti.'caagggaaagsgecaccctctei:?gcagggcea0tc3g8§igtt<!gcagcraeua>?ccis;suicca<u'agaas£· et£gccaggcitv:eaggetcatca!ei?i?s«iges?eeag.cassgccactg2cafccea£acKsaita}atsacagigggtctggga caeacatSiKOxtcaceaicageagaei.ggagangaagatiu.gcagtsuutac!gteagcagcgtage.aaetggcctcegacgticg gee a a gggaceaa g gtgg a a sttca a a

S3 arenas acid mqncnee (SEO ID SO". IO iim lading signal pepdue in usKiedmed UahcsE

m. 4f. \()i U /./ /7 QSR\ I LSLSPGfciRA 1 LSOR ASOSVSSYIAW YQQK

reOAPRLUYDASS R A TGI RDR FSOSO SC-TDl·” ΓΙ: PLSR Lh FHDEA V Y YCOOkhNWFFTF GQGTEYEJK ¥> amino acid..sequence t.SEQ ID NO: 1-9/ excluding signal, peptide:

EiVLTOSPAT!S!:SK^RATLSC^ASQSVSaXLAWYQQKKv«OAPRLLiYl>MSRA ΓΟΙΡ DIG (SGSGSGTDFi LTiSRLEPEDF A V'Y YCQORSNWPPT ΓΟίΧΤΓΚΥ BIK.

Vi CORl (SEQ ID NO: 20k QSVSSY

V; CDR2 fSEQ ID NO: 2D: DAS

¥{ CDR3 fSEQ ID NO: 22): OORSNWPPT

Vp mseleleamd sequence (SEQ ID NO: 23?

Aigaasutgggeiga actgggutti:cn. gimen. umnigaggmtcaagtgu. asgissuacisgtgsagictggaggas,sg am g:ccagccWggaraicccWce«cactcc:gfai:ascg?csgg:;iicacrrcjg>agci;Hgacaraca^mssiccgCi,asgciCC agaca.iggggctgg.igtgggtggeaguatrossaatgarsgiiusfaataaataciutst'agacn cam asm a see a;° ncacc a a; Kwagagacanticcacgaactcgi.:tgt:Rdgs:aa>.u.g;iaeagcc{gaaagccgaggacaogai,tsugfarc'.ngtgtggaaagaac ; aci a arc t: a a a cat.· m a a ac c a g a a a acc c tg a tc acc a d teen, a V·., arums acid sequence uSEQ ID NO: 20 (indading signal pepikle in underlined italics}:

Λ/Α7 '< US mi 7 Vv\/./ //(/tl/GOYQ!./VESGGGYVQPG R SLELSCAASGFTFSSYDiHWVR

OAKlKXILEWVAVIWNbGSNKYYADSVKGRFnSRDNSTNSEELOMNSLR.AEUTAVY

YtAS:K}T;A(3L!;d;tWI^i:;T!.A!1A;SS

Vgamino as.id sequence <$EO ID NO: 25) excluding signal pepudet

QVQJ ..V KSGGG VYQPG R SLRLSGAASG FT FSS YDi H'W V RQAPGKGLEWY A VlWNIXiS NKYYADSVKCSkF1'(SRDN;STMSLFL0?vI MSLRALDTAVV YCVGGrADLEH¥>; DOGTL vivss .....

V» CDK i (SEQ ID NO: 36): GFTFSSYD

V;· 0DR2 (SF.Q ID NO: 27s; jWNDGSNR

V<s CDR3 {SEQ ID NO: 28): VCXQ^Dy^HWOQ V: rasa esc acid '<ecjuer.ce $f:Q IT) NO: 79;

Akugggceuegeu:agcu/rsggggcSceTgi::gc;ciga:aeeagg?gccaga?g:gsean:cas;Ugac;a.agaTccasccH/ ac:!jieigcatctgiS|:gra:aaagagic;K:cufcacnsresgacg;:gie:a:^ya:uiU!eeugcigiUU4?c.r;ggiu:eage;uj.eaie eapagaaagecccuiegicecrgaietyigeigeafecagttigeuaagiagggn ceeieaeggneageggeagiggsnucgsgac a£atae3ete!caceaie::geaaecigc;igcc.ugu:n>jiingc'aaertauaei£e^acagtatKafaguaeeeieieaunucggci.' »?tg»gaecaaggtggagateaa&amp; V; amino acid sequeaee (SEQ ID NO: 30's (including signal peptide I π underlined italics):

mVL \QJJ.(TTYURGDiOVnOSPSSLSASVGDR VTH'CR ASOGISSWLAWYQQ KFi:iKAi>KSL)YAASSLOSG\3>SRrGGSGSG'rDiNj.:riSSi.X?PhDi::ATYYC<:x:)YNSYPLT egggtkveik

Vj amino acid sequence - SEQ ID NO' 31) excluding signal pepGk;

DlQM'f QSPSSLS AS VG DR V TITCR A SQG1SS W LAWYQQR P! · ΚΛ RKSU Y AASSLQSGV PSRFSGSGSG'rDI-1l.;nSSLOPEDFAlYVCOOYNSYPi:rR3GGTRVHIR

V< CPRi (SEQ ID NO: 32): QGISSW

V, CDR2 {SF.Q ID NO: 33); A AS

Yi CDR3 tSEQ ID NO; Us. QOYNSYPi.,1 JFS \ H Oil 3-33; 07-27; JH4; \k uficieic acid sequence tSF.Q IT) NO: 35 ,s \Taaugur gggeigage igggt lucctcg u geie-maagaggigteos tgseag aegeage igaiggaatet agggguggegt g g tc ca ace? ag gug g ax e ip up a; ren.v igucugeu;c:ggaUeaceneng:agiie;gaea:gcaeigggtecgeeaggelc i.nggcauggggetggagtgggtggeKguaiaigg?atgKiggaekuaumaineiatgi aaaeiccgtgaagggecgniteaceat eievaaaaacuaueeaugua;.;KgU';gf:neH.cai:^ega:K'ngccigagagccgaggaeaeggcigtgiaitacigigcgaguggf agtgautaagaggUtelugauaetggggceasgguOceetggieeeeiOeteciua

Yh amim* acid sequeive (SEQ ID NO; ek tint.hiding signal peptide su

ΜΕϊ'Γ,ΜΧννη. VAUJifiVOi ΟΥΟΙΛΈΝΟΟΟ VV0P0RSLR3LSCA ASORTSSVDMj PAY kQA)X>KOLEW\PAviwYDGSNKYYAD$VK<iRRlSRDNSKNTLYLQMNSiJL\EPTA V Y VC A RGSG N WG PHiY WGQiJi I. \T v'SS

Yn uminu acid sequence (SEQ ID NO: Xn esduding signal peptide:

QYQi.Y ESGGGY VQPGRSLR LSCAASOFTFSSYDM HWY RQAPGKGLHWY A V TWYDG SN K.Y V A DSVKGRFTISRDNSKNTLyLOMNSLR AEDTAV V YCARCSGN YVQFFD YWG

QGTLXTVSS

V;, CDRI (SEQ ID NO: 3S..s: GF-TFSSVD

V;· CDIG? {.SEP) ID NO: 39): IWYlXjSNK Y-i (T5ROSI.0 ID NO: 40): ARCi$(BMii±Dl %. nucleic acid sequence I SEQ ID NO: Ap

Noapppic(:u:pi:ic:ipva:cipappcict::pc!pilciiaaccc:;ppfpcc5pafif0acsa-cap,4i5pa eongteiceaieen: ;v^lg:coc;nctpiappa£aca3apaaicca;ca>:ns:cpppcgaaaa:pspt;np5 24:appo,3u:0i'aipi;::.auapcupaaac i:ja;:paaapct.ccPK:picccOaKPaEgcOianca5paoc3a:;pU£pSK:Ci.;nc;5aap1a':;pi::;pc;nnia:;acippa:v%. asaute acn: teaeciuea sc agcctgcugoeq: aa pan:igcaaeunriactgeeanea giutaai acuaccoicg sat gneggee a a a.gg acc an g a t a a aa ate a aa V; amino itcad sequence {SEQ ID NO: 42; {including .signal peptide in underlined IraiicG;

MRVIAQf WUIU1-PGARt DiQMTOSPSSLSASVG!>R VΤΠ CRASOGlSRWtAWYQQ KPEKAPk.SIJYAASSLOSGVPSRF$GSGSGTDFTI.TiSSLOPb?-)FATYY(XK)YNTYPRT FGQGTKVE1K

Vt amino acid sequence {.SEQ) ID NO: 4.0 csdndins signal peptide:

DiQMTQ.SPSSLSAS VODR VTITCR ASQG 1SRW LAW YQQK PER APK.SL ί Y AASSLQSO V PS Rj; SG SG SGTDFTLTJSSLQPE DF AT YVCOOY MTYPR1 PGQGTKV EIR

Y} CDRI -SEQ ID NO: 44;: QG1SRW

V, COR 2 {SEP ID NO: 40: A AS

V: CDR3 (SEQ ID NO: 46): QQYNTYPRT

Yn nucleic acid sequent. iSEQ if) NO: 47)

AlggagU:gggugaguigggimecrcgttgeiciUuugu£gien.:eag;£ieaggtgeas.C04teDiPt tgsssgeggegi ggleeKgcugggaggtcectgag:KistecigU£cagcgtetgg:nkcn:eiri.nu{.nenngaeargeuctggglccgceaggek' ece:ge:iaggggeiepaocaptgpc;iginit;ViSgiatcuagsaagia.;ic-\.c;aenuge;igncieegtg.ugppccfta:i:caecai viciiv'taeg^\^ycu{i>act.u.iiiff&amp;i:v.c.ij»s;uca:i^ak”^-ciriciccica V„ ammo acid sequence (SEQ ID NO. 4K jdndudum Signal pepade in urnka Sined Palies;:

MFFGISWVFt I'M l KYrPQlT>VQLVHSGOOVyGPGRSl R I NC A ASOKITN] Y4)MH Vd VR QA PG K( 5 LEV* VAV iVVYOGS.NO¥YADSV KGRFTiSRDN SKNTi ,Y\ .QMN11 R AF.HTA V Y ycargthw o vfdy wotxrn λ tyss V’,; arriiuii acid sequence {SEQ ID NO: 49; excluding mg mu pcpndc:

QYQE V ESGOG V VOPGRS LRLSCA Α$ΟΓΠ:Ν f Y DMH W V RQARG KGI..HW V Λ Y IVv YOG S NQYYADSY KG R FTISRDNSKNTLYi ,QM N (LRAED'f Λ V Y YCAROTH WQYFD V WGQ (ΥΠ. V'TVSS V;f COR 1 iSF.Q ID NO: 5id: (l.F11;;N!Yid Ya CORd (SEQ ID NO: 5!): mTDGSNO V„CDR3 !SEQ ID NO: 52.: MSIBISSSY IHgVKIVKID-ifeJKl) V; nucleic add mapum-ce <SEQ ID NO: da'?

Atgagggicctcgcicagciccisgggctcc^crgcictgracc'cagat^ccagaigioaorccagatgacccagtctccatccic aci£tct%metgtaggagac3aagieacc3ic3cujii.cgggc»agtca&amp;gguitia£ea«ctggtlagcctggtaieageagaa:«: caga|>aaaytxxciaa«ictxH«atctstgct.?catccaaitigcaaagtg2ggicccatc;iuggtrca^cogc£i»r§«atci5iggac a5jt{tc*t»:?ct{-acc;iica-ica,sccrgcrigcc{g3agai{u»c33c«a{uicigccijacagiafastagtiacccicg3ai.'giicggcc «a»-«v'2£'C-3ap.gigaaaaTcaaa:.

Vj a.nlno acid sequence (SEQ IP NO: 54; (including Agasi pepridel^und^ned;litaies):

.•V/RrOO/./.O/i./i.CWa^CPIOMTOSPSSLSASVriDRVTrTCRASOGISSWLAWYQO

KPHKAPKSLIYAASNLOSQYRSRFsGSGSCiTDriLriSSLOFjcOFAfYYCOOYNSYPRT

mQGTKYEIK \'t amino add sequence - SEQ ID NO; 55' excluding signal pepade:

DIOMTOSPSSISASYGDR V) Π GR.AROG(SSWl,AW YOOKPEKARKSLsY AASNLQSGV PS R FSGSGSGTDFT LT ISSI .QPLDFATV VCOOYNSYPRTi GQC HR VUK

Y,uCORi:. iFQ ID: NO; 56 c OGISSW

V; CDR: (SEQ in NO: 57V. MS

Y, CDR5 (SEQ ID NO; 55 ?: OOYNSYPRT V'jj nucleic acid me. am me (SEQ ID NO: 59? A{gs:nrarg^.ciw3si:igg§utK-ctcgtiacutHtaa^aggi^tccaaigtc^|;f^cascrii!ag!£agici««gsg3g«ceH) .stcx”i^cctsg^3gs.'tccct^seairK'iccigi.>;t.‘;)Si.'gicf5f-;':iiica«ctiCLi»r3siciuiff"catacaciagiftccgcciisgv;tcc ;i;>iYcaa2a«'acte|ia2tg.s^tf;j.K':A:'Uctat^gLiiaau^UAgcc;na;j^g;Kn.iigc;:igac-ccgtjiLUiggiicc§;rac^ccstci ccaj«u!?yc;uricca;^s;}ac^cgc{a2.arc\sc^3igaiiC3i>coisaagagv.'C»rassacav^*iCM>twttaci|;igc|raa:}gas 2Slit3^a^iuccr£2icr^naaKaodc^aitacUi”wgoc«hi2c;jircci"2i£r3Ct{ii£riccica V',; ammo acid sequence : SEQ ID NO: 60s i including siamu nepmle m underlined uallos);

^l67υ^NUOO-7Λ¾/i^r;Oί^υVOLVESGOGV^ΌKJRSL¾LS(:ΛΛ^GFSFSSYOMl·IWVR QAFGKGLFW V A I. LWYDGSBK DFADSV KG 1¾ FOSRDNSk NTEDLQM N S L.RAED'TA y YYCAREOLAVKsHW’VTm .WQ RGTL VTVSS

Yjj and no and sequence ;SF.Q ID NO: 60 oadnding Naum qepqde;

OVQLVESGGGVyppORSLKLSCAASGRSFSSYCFMBWYRQAFGKOi i-WVAl.l.WYfXl SHKDPA DS VKGR rrtSR DNSN ΝΉ ..DLQMNNi .RAEDTA V Y YCAREOE AV KiHW V FDi. WGRGTLVTVSS V; CDR1 (SEQ ID NO: 6.?.n GFSf SSYG VO CDRO iSF.Q ID NO: 63i. i.WYPGSHK VM CDR '> <SEQ ID NO: 64}; AR FGLAVPGH WYFDD IG5 V K {V K I -10; j Id s %.nucleic -addSeq uende (SEQ ID NO; 65} Y%fgasgOcccc^:o;cagck:o:ggg}>;UasgcigcK;oagcic-acagxunccagai2t2ccasoc3i;u2acx.ca2u:tccau etc cilaicicoa;<ogtaaaoeacaaags!n)cc.a<:ac:taocgoa<::n.ig7oaggaoasiagoags.ootnaacctgaaa:t;aacagaaac caaamumaeiccm:nmecTnneuua3saccieca(msgaaayOGODtVC:6eaapaucaaeanmpmnm. raeaac a a mm 'acts 'τοη<. *. ateaseagoctm. a sretgaaga tin ge.tacUuttae· ateauea a s urn: me n aeee te g a ac * n c a see a negg a ecu a a a t a a a a a re a a a \q arm no and sequence (SEQ ID NO; GO (including signal peptide in underlined dalles); {»7 /./ /. WLPi >V Iff CAIOLTOSRSSLS AS V G PR VT1TCR ASOG l$S A L AW YQQK PGKAPKLl.lYr)ASSLESGVPSRFSGS(:}SGTDF11.:nFSL,OPjdDFAlYYCOQFNl'YPRTF GQGTKVIGN. V-. amino acid svemenet (SEQ ID NO: 6?} exehOmg amnai peptide:

AlQI.TQSPSSi.SASVGDRVI Π (^ASOGISSALAWYOOKPGKAPKEtlYPASSyiSGYP SRImGSGSulDEi LTISSLOPEDF Ε'ΤΥ\'ί'0Ο6\Τ YlEYTRdOvlTKVIriK

Vi.CDRI (SEQ ID NO: 6S>: QCiJSSA % CORE t$FQ ID NO; 6¾ OM:

Vt CDR4 (SEQ ID NO; 70); OOFNTYPRT 2G9 Μ) Ο 0 303:1)07; .1114}

Vi; .m;cle;c add sequence <Si:Q ID NO: 7! >

Aie;;e;en!qggi:t;reecn;q::nern.CiCvuejeicc:rcen)eegi;Vcon;Veieai\eneonqrras;tas;;iS;C:^cs^qacei:iq,c ate mm cc:qo.';ceic.; ameeacmtccmteenpepsemaoummcmeaqtaeamnmcaueaatpaatm eecaasctc eaum.sagss^eeiin^aeiegiy^esseatnnac^qeaeisaqi^isere'-seeeene·. uunmpocm.;npaapascemumacia c?cc;i;rec^;cutecra.,sescH.;'cqyCiSciqo;ussiga:e:sc<-cq;ceeyecveaeeac;a-^;3ct£;iacereu:efeTaes;ig«;:·.: ch'ci.:.·secuva«cact?mcea» a oaac-.Λ naana».eaten. etca. <h' v·· V-· V·' \- V-' y.-.-i.-.'jv.· V-·· ·.·· V-··· V;; asm no acid sequence t'SF.Q ID NO; 72) tinchidhte Asmal peptide hi underlined uaheaa

Mf/ iU.SWVl·L10/,/AY;\V(.'QV ΡΙΛ l:SGGOVVYvHXiR$LRL$OAASGFFj SKliDniWVR QA PONGS r.NVV NViw\O(.iSNKYVAD.SVKGRFnSHON'S1'N,;<Li"L0M.N'SU?AEDT'\VY VCVROTAPliiHWOQChr LvjVSS V;i ammo acid sequence (SEQ ID NO; 73) excluding signal peptide;

OV()l.Vr7N:iOGVV7VhAd<SiJO^(7AAS<:nN1.mSjlDjj-)AVROAPOKGlJaWVA\ijWNf>GS

NKVYAD$VKGRFnSRDNfSTN$DrLOMNSLRAEDTAVYYCVRGTAOUr:i-lWD(Xr»TL

VTVSS

Vn DDK; iShQ 10 NO: ''An Oi;ILj>ML> VO CDR2 tSEQ ID NO; '75)71WNBCiSNK Vn CDROSHQ ID NO: 70: VRqri Ai)[J;:||WDO 2G9 VIUVK- ID-lfr; 1K4) V; nucleic acid sequence i SEQ lid NO; 77 \ \i p;m a Acetcpaeaqcacetpaqacn. eqaepe·;. ctgmeceaaei acc.igm ataaenmeaqco gacccagtetccatccrc

Iuopietpe;ricrpl;qtaapae;4niSteaccatCiVUgtcgqqscaaptinqagpaaifoscapct”ptt;!s;:ei.!;pt;iU:;.ipeagaa.ac c a a a a a a; s p cc a c: a a a: ec<: taaietumctgaaieejausgeaaagta\iaOcceateaaaptea|iepgeaniggateiggseir ;n>;:;iUeac?eKaaeeasc;qecagccfpcaaoc:e;amnUCqaon;;.cHa!i;Ka\eio:aaca<uataa:aetnmxs:leic;K-UTCS'gcp; aaaS'mmcusmgfggamfcaua V; mourn acid sequence tSEQ ID NO: 78; unctudinu Ngnai peptide in andesnned italics);

MRVLAVi HUJ U CiPiim DIQMTQSPSSLS ΛSΥΠ1 >R VTITORASOOISSWLAW YQO Krd)KAPKSLIVMS^i.OSG\17aRIGGSGSOTDF11711$SU>iAEDi:37A'Vr(X>Y.NS;OiU:. FOGGTKVhtK V; amino acid sequence (SEQ ID NO: 79) esduding Nanai peptide':'

D10Mr0SPSSLSASVGDRVTnT:RASOGiSSWi.AWY00K,H:LAPKSUYAASSi-QNGV

PSiO-SGS(:iS(:rTD10'l.TiSSLOFEDFATYYCOOYNSVFLi~rOGOTKVLTK

¥>. CDR: uSFQ ID NO: 80): GOSSAN

V, CDR2 iSEO ID NO: SO: A AS

Vi Π no ID NO: 82): QQYNNYiN.'T 'V&amp; 831

Vggasitteegcl gaaetg «stutfecti. aimed. t maa:ing:A?0· ea<p gu. jggi seageigg tggag rciggggga ggcg? gpi:0avct?i;g;:sgijiccc;g;'!iacictcc!^igcai;csicti:^inc;u:i.uv:';!a:cun:!ii:gc:il 9i:acir:r^u.<:g<.:^\^yc?v: ca*jgCio^^^Cv^'s.^^4sy”fgh’c:i;)Ujui!.;;’t;)ig.n^aaf.,t3Ui:i;'tMi:ii.'Uij.,caff;!CsCcgi£r3aai:gcoaaucacCv'ii otcvagaaacuaUΛau^au<.acgct|L'μ:nct;*o;uuμ;wt\ίgcoq^Ja;JSi.'o^.lϊioa^.':iCiΐffεrlϊϊ^,Uitιuct{)£¾>'ϊ>aaαga igg.dggacineiatgguegagg:uataatau:ngatatciggaaecau£gguea:Ugg?eaeegi; icdea VEf amino acid sequence uSLQ 10 NO; 84; nnelndma signal peptide in undeslmrd

Mi-TOi.swvn va i ΐβ(; t xx ό v qj v fsggg vyqpg r si ri ,νγλλ νορπ-νηυομηυυ

ROAIC<KGi?i;WVAilWVix^S,\K.YYADSVKGRFT.ISRD.NSK.XTi..DL0V1NSLRAEDT.\V \ NGAROGSVmiVRni. Y\ 40IW (4i( ·ΓΜ\ Ϊ YSS V,\amino ockl sequence iSEQ ID NO. Sid excluding dgm'd peptide:

OYQLY&amp;SCGCiVV'QPGRSKRi SCAASGFTrSI IYOMI i\VYRQAl*GRGFEWVA(IWYDO SNKYYADSYKiiRFrKSRDNSKNlLULQMNNSLiU£DrAVYYCARrXiSSaT\I\^Gl.NV FDIWC<X3TMVTY.SS

Vu COR I iSF.0 ID NO: 80»: OOISHYil

Vn CDR2 (SEQ ID NO. 87s: IWYPGSNiv Y{! <ΌΚ3 (SLQ ID NO: 88»; ARCfoWTTMYRGI.NΥΓΡΙ

Masmsiiaa

Vi nucU'ic acid sequence ISEQ ID NO: SO)

Aigepegicc;eeeicapc:eciappae:eeiaetpe;cipinceeaaei£ec;<p,ite;aac;!iec,!p:;:e;ic:ec;ieH:u.i.;ueiie acietciei:'.uei£iaaeapec.:p;!aK;aci.\iie:)i:i;p>i. ppp;.'paa:eaee.;U;iuave;ii?cG^dapee;ee;;eiaeeae::-;;se cagaeaaapeccctaa2iecc;gae:t:upe!i:< aioraaufgi ;j e; i pi e eg e: e-;e e u.\s;; e e: ;ca a c a a c a e: eea til e ρ ρ a u a^a:ti;a.Kt;:icai.'eaii.\5geapccti)eae,eeti)aav'adib.>.caaeitidt3:l£ceaacagtafaai;ipttac-A toe-: aeetu.ggi.e aa|i«gae:ie:£aetp:^n2iitraaa V; amino acid sequence iSF.Q ID NO; 00» t including. signal peptide in undet luted italic;;):

^^4i(^.U^XU3:7;P(MR<:P(OMTOr>SSi^ASVODRVTnCRA5IO[)^I.A^TQQ Κίΐ N KSi % \ ON 03< N I^KI N<.N(.SG I D! Ο 11\N 01 ΜΗ \ I Ί s Cv^ νΛΓΙ'ϊ EGQGIRLE1K \'i amin-ί acid veqnenee (SFO ID NO: 01»excluding signal peptide;

DiQMTQSPSSi .SASYGOR VTiTCRASOPISSWLAW YQOKi'EKAPKSi JV AASSLQSG V PSRFSGSGSGTDFTI .TINSLOPHDFAT VYCOOYNSY FfYI FGOGI RLEJK.

V, CDR l (SEQ ID NO; 02): ODISSW V{ CDR2 (SEQ ID NO; 93): AAS V; CDB3 (SEQ ID NO: 94k QQYNSYPPT A fas bar b.am chans νο;.- aNo ioustd to be active ;o follows (though only the above bah; chain s3RS-1 Bl I VK #1 s wav ua’d ίο ihe above Examplest V; nucleic add '-equenee «SEQ ID NO: 95) A tgaaaau'i to go tea act cava agettetset aetet g pc: e ecu a a (a a c; ο:; ea ΐ a c e; e cc > t gt t euoceaioctct. atccte i. c::iie;aaauoo;a«g;;o:;cjaaaieaaa,na aei^ccaaacaaaK'aaaiaaanugcaaiaeUfaaaciypaajcayvaiOiaac caj^uiauaecevviaasts-xctsateiat^ gaend ca;3eiaca.^g:aaa;AiA;e;Ovaaaauaa.aeageaaOva:0e;aaaac ;iaaeoe;0acicaccakva\a;a;jtciacaaca;aaaaatn?;ai.'3acU'an;&amp; igeeuacugdoaatjunacevnweucutcygcc uaa gg;u;acgacta p aatteta

Vi and no acid sequence (SEQ ID NO; 96) (including signal peptide in underlined italics):

A iQi-T QSPSSI dAS^IDRYTiirKASQCHSSALAWYOQK

PC K APKSLI YAASSLQSG VPSRPSGSGSGT OFT LTtS$L QPEDFATY YCQQVNSYPPTT GQGl'RLELR νό amino add sequence (SEQ ID NO: 97) excluding Mgniil peptide;

AIQLTQS PSS L5A5YGDRYTITCR ASGGISSAI.AW VQQK PEK APK SI. IY AASSI-OSGYF Skl-GGSGSGTOFFUISSi .OPRDPA3 Y YCQQYMXEOYGQi V) RI..EIR

Vt CDR I iSRO ID VO: 98): QCISSA

V; CDR2 (SEQ ID NO: 99;; AAS

Yj CDR3 (SEQ ID NO: 100»; OOYNSYPPT 31112 VII (VII 3-33; 0707; JH4) V;; nucleic acid sequence {SEQ ID NO: 101) aipaaad)aaae;a^iyc:aaad)KTtegi?avje;tuaa&amp;aaafvK'C;iajpk;aaatac-iec;aaiaa:;ao:;aapgaaaaaaia atccasi^tegg.iggsci.'iUgai.eicieii.cictgercK-iViCigeatieuvxucagtaae^igaeatgi.aetg^vSi.t.aceaggetci: aapeuaaa a a a: a a a a i a a a: a a*. aa? tats igaoo saiggune t;a a etutuci utgea guctcv. at a aa aa, aec aadeaecaoct 4aaiaaaacr!;u;acaaaie;eacpctatan:iccaaataaaaaaccsaap.ia,iev;iae;x'acgpe!p!giau;K;U:aveaaa.aaia a is staact ggggfnesnaactactggggceag jigaacccs a a tv aecgicieetea Y:h atAsao aeti sequence (SEQ :ID NQ: 10¾ (iaelBdlng signal peptide in itndealised Saties;I:

AtE/?)/AlP\YAK4//^;VYA0Y0EVESC;GGVV0iAiRSi..Rl.dCATSG.ISI.FSSYDM(dWVR OAPGKGLEWYAYIWVDGSNKYYADSVKGRF riSRDNSKN rLYLQMNSLGDBDS AV Y Y(:mi^iNMSTO Y WGQGTL. VTVSS VS5 ami.no acid sequence iSFQ I'D NO: 103> evdudina Ygnal pepiuic;

QV QLYFSGGGWQIX i RSLKLSCAISGFI l-SSYDMH W V'RQ.A PG KG i i:WV A VIW Y DG SNK V Y AfXSV RGB FT IS R DNSK.NTI . Yf .QM N SLG PP.PT A V V YCARGSGNWGFrDVWG QGTLVi'VSS V31 ("DR i ;SEQ ID NO: 10-1): VdTFSSYD V;;CDk2 tSFQ ID NO: I «5): [WYi)GSNK V.i; CDRo ISBQ ID NO. 106·: AljGSCiN WGrijDY 3HU.VEMffK Ift-lfc JKii V; nucleic acid -^.CjUi.rK e (SFQ ID NO: 107 * A:paca«ici.ica.cii:agcia:ipacgciccfaaigcicigiuacc5paipcc-aaaigiimcaicc0aaipaci.i:aaii. iocaicaic aci2tciacaiiapiaaaagaa'aaagic:a:aaicaciipicaaaaaaaAaaaaiaiiaacagptpGiiDccDptaiC3gaa|:aaac ci^!AiKUjaiK\xt'\ai‘4'iccctgatciatgciscatccajtiigcn;i»§tagj:|»tccc»atcaa:ag«t:agc«gc^gq;gijtct«4>8ac aparncaaicii.aca:0capc:an.cigc5accmaaaaiiiiacaaci:aiiycigccaac5giaiaafac?i;!ccck:aaacpiicp:na: nag ggacea as gtg gaaa&amp;aaa· \A amino acid sequence <SFO ID NO: HJS; (including signal pen ode in underlined balks};

m\Ί, \ QjJ <: /7./ U.( Ί Π a/R'DIOM IQSPSSLS AS VO DRY T JTORA SQGISRWL AW YQQ KPkKA PK’sUV AASSLOSG VPSR FSOSGSCTl PI 1 LTfSSLOPLDFATY YCQQY NT V PRT FCiRXVTKYi-JK. V; a ini no and sequence (SHQ ID NO: 109-) excluding signal, pepbde;

DIOM*TQSPSS?-.SASVGDRVTITCRAS<XtiSRWLAWYOOKFEKAPKSLiYAASSLOSGV

PSRFSGSGSGTDrrLTTSSLOP£PFATYYC!CK>VNTVPRTFGOCiTKVI::lK

V: CDR i (;§BQ ID NO: i 10): QGiSRVV

V·. CDR2 {SFQ ID NO: 11)): AAS

Vs CDR3 -ISBQ ID NO; i im: QQYMtlMl; ALIGNMENTS ARE SHOWN IN FIGS ir AND 16 1« Vivo Non-H«man Primate Study

To assess tolerance M a ad human CD.;” mAh HFS m non-human primates. .¾ evoomolgus monkeys were me a ted with one Lv. oSe-e of I. < or 10 mg/kg of IP5. Animals a,em tbllowed for 2\f days, Total lymphocstes (bused on «Ide and forward stouter si-co, memory B cell·. (CPdOs- and Cl»5 bright s. and tnonoeyies (based on Mde and iorwrud scatter si/ct were sp-ined with atm-htamm ly.G ami bods (hold Ones and compared to u η-.tamed want mis tshaded htstupram). The results am sh-.mn ht ; Dure 17.1 Owe:· results 'how that the 1Γ5 mAh w a a bound u> the surface v-f eheuiathtg K mprw;. yp> brum tr> eκpress 0)27 lor the emnv duration -..ft iho t-mdv Monocytes, cob-: that do not expte;-s CD27 did noi have IPf Mndmg, hi addition, to de-ermine the edect of I F5 on circulating l\utphcx.yie populations, lymphocytes wurn stained wish Mbs.set mat hem and the D positive odis plotted vs time tit Figure 17 fot each attmtaj tteu-ed at the diHerem do-vs t square points - I mg/kg ; d red: pen it'o — χ mg/kg; triungh.· points .-- Up rng/kg). Re‘mbs are shoo, it ttt to pure lb.

Col Get i\ eh, She re-.ubs iron: those studies, shoo p us Figures i 7 ,md id, deutonsnaro that IF* was well tolerated and did not \lgmfu.awh deplete chettiating lyrnphui.Yles iMthet titan some trau.heut dcpletiOit oS Nis odist alter a single S Id ma/ks d='se Its addition. there was no ok'satioti tn body temneramres and tie- detectable level- vi T7sF·t*x. 11 .-b or Π.-1 μ.

ArUl-CD27 nsAb.s Enhanced Anttgeo-bpeciSc CPS-r T-ceH ProUferatton and Acttvaikm. PentarniT Staining on Mouse Peripheral Blood CGB and Splfnocvi.es

Human CD2'? transgenic mice (iiuCD2?--Tg! were ramus enmn h Injected with 5 mg of chicken ovaibumiti and the panel ol fully human atihbodies recogmeang human CD.:. 7 generated as in id ample I t'GD27 human mAbs). Rat anti-mouse CD.: 7 clone A7 124 and irrelevant human IgG i were included as positive and negative controls, .respectively.

Each antibody (250 t.tg) sva« co-injected with ovalbumin on day 0 and an addiiionai 250 pg oi antibody alette tm day I Peripheral blood and spleen cells were harvested on day 7 Spienocyies (lx FT; or whole blood (1-00 pit were used for staining. After Pc-receptor blocking, cells were stained with 1-0 pi of H - 2Kb/S 1 iNFF:KL a tetrameric complex or mouse MHC completed with the peptide T cell epitope irotn ovalbumin (Beckman Coulter), tn a similar penUtmerk complex (Frontironstee anti CDS (eBiosoiencc; and anti-huCD.?.? mAh or atsti-msCD27 mAh t'BD Biosckmees} at .room tetnperature tV»r 30 minutes. Oils were then RBC-ivsed, washed ansi fixed, and at leas! 10-0,00-0 events were acquired in Flow·' Cytometer I..SB CBD Bioscienees0 The traction of tettartter- or pernaoier-positive cells in the ODB' of 00027' gated population was determined. Rest;its are shown itt Figures |0 atul 21 from which it can be seen that the ami-€D27 antibodies significantly enhanced immune responds.· F4JSPOT Assay

Spiettoeytus (2.5 \ 10' as id 0.5 x 10s) from the pcntatner stasning preparation of Example 8 above were placed un an :mf · iFNy owes wl seed antibody t rn.Ai.n coated plate in iApIeate wells, after EBCafyAV SlINPIaKL peptide was added at a final conceit iraf mu of 2 pg/rnl, A fesdgptip.hd· .ebp&amp;tRfWSisi Aeienp'l&amp;t eae:b sample ia tripMeate m the. .absence of peptide. The stored aeon was maintained at 57''' C in a tissue culture incubator ovenughb BIJ.SPOT detection was performed using an ELiSPO'T kit tBD Bkwdences) tVfhovvhig the immo lecturer·· protocol. IKNy-spoi number was counted Rest·Its are shown in Figures 20 and 21. iVotn winch it can he seen that and 43)27 nsAPs sieciileasnly enhanced T ceil activity,

IsiMkJl

AiUi-0027 mAh EntotMes T celi Responses to a Vaccine Antigen ft)Q727 OAhsgeaic place :wera Immunized with Spg 1¾¾)).of the APC-targeted v cine comprising an antontou-,e DEC-205 IgG antibody fused to os alhmnin tOVAs uefeiTcd ? * as « mDbC 305-OVVc In cornbinahou o ith the :44:1-(3.)27 human mAh 11-5 n.p.'i at ta-iOns doses ids. 5u, 1()0, 200 or dot) μ») One wueA later spienoeytes were analyzed for cD8- Tcell maeuvdy to the OVA SiiNFEKL peptide )0 VA peptide 257··· 2.64· by tcirauur staining (0 h.-tramer positive ;ili CDSs- shooui and IFN-y EL IS POT bs the piocctlnre as general!) described in tAarnpies 8 and 0 respectively. The res tuts are shown in Figures 22 A-O. wherein Fipu?e '22A shows the protocol used. Figure 22B shows the results of the letramer sunning experiment and Fiacre 220 ''host s the results of the 1FN -ganere ELiSPOT experiment. These results indicate that the eo-admmt stored human mAh IFF significantly enhanced the I-cell response1', to the vaccine component adutinAtered.

Synergistic Effect of antiA'T>27 ntAb flFSutod TER Agonist (Poly 10) on T ceil Responses to Vaeeine {anM07BC2f)S-OVAF huCT)27-Te tt ran'.gemOt wet wild-type tWTt mouse lutermates were imrapemoneafly injected auh usttt-CD27 mAb IFF )50 pg? tjn day -3. and then subcutaneously via 4 paws injected with tun t) - m 1.7 e c 305 - Ο V ig ggt pins Poly 1C 0. 25, 50 or 100 ug ο;) das 0. Spleens were collected un day 7 and a>tses».ed by tetramer staining, lENy- EOSKjr and ΙΡΝγ-iCS, Mean a. SU of positive IFNy-ICS among gated CD8 1' cells from 3 mice per poop were ealee hard and a panel of representative Dot Plots was collected. The it*-.alls. shown in horo 28.A-D. indicate that the the anti-CD27 human mAb acted sy tiers isticaik aiih T1..R3 agonist Poly ΚΓ to enhance the T-eeil ·ο*ρο·νθΝ n.· the vaccine component adrnt nisiejvu.

Example 15

Administration of a nil “CD 2 7 mAh Prior to DecSOS-targeied Vaccine hi die Absence or Presence of TLR Agonist huC TC'· fg mice and wiki type ; vVT h derm sic·-· v, et e tntmpei itoneahy injected with amM"T}27 ni.\b BOiggi on saiious days related to raceme as indicated in Pie me 28. and subcutaneously via 4 row s injected with and-mPecdOg-OVA A pg; pho or minus H..R agonist Pols 1C PC <20 IgA on day A Spleens were cRiveted on day 7 and .wsussedby tettamer staining and iFNy-ELLSPQT. A representative panel ndeiouaicisstidtd:a|: among gated CDS T cells ts show n m Flgtirrw 24 and 23 IFNydiLISPOT showed a similar: pc item.

These results show that, surprisingly, v- Iscn anti-PD2? antibody h administered it; eomblnntiou w ith a saectne its the presence or absence ot 11..1¾ agonist, TwreH ;u.{ivatiun is greater when tite antibody adrmnisiemd before ;hc vaccine, tor example a day or more before the saccitir tantigen 1 is administered. pAmiipio id

Atdi-C'02? mAh Combmed svith TCR Vrtiyotkm Aeihaies Ύ-cells from .Human (.'.1)27 rrrmsgeoieMiee

To assess the T cell activation capability of IRS mAh. I cells were pun lied front spleen of hCD27-Tg mice bv negative select ism wmh beads. CVils were labeled with CFSK and incubated with i.’Tggdu! ofam!-(21)27 human mAh 1F3 m tsotype control Re 3 days^ l^e'orossdimkipg-MfRfeptPltti· Ig<3 wag passed: fli.rbi.tgh an endofoylu removaleolimo hefoi-o use. IFNy-lGS; and €SFE ditedoiromong CDS and CD4 T cells « ghowti in Rgure* 26 and 27, TNPa-li'S showed the smile pattern as IFNg As shown In Plgum 26 and 27. w hen Combined with TCR activation 1Γ5 rn Ab effee lively Induces p-oldvration and Cytokine production ttom V cells m uuo, The data show this? both cross-linktn» with anti-human 1»C and T cell receptor activation ά ith nntFCD3 mAh were iwqthred for IfS tndtteed prohiemtson and cytokine production.

Example I? AMF€T>2? mAh Enhances the Efficacy of Vaccine In a MCM Melanoma Challenge

To a-;sons in r<Vr> antiuunntr activity, huC'027-Ί g and WT control, 8 mice per group, vac it subcmaneoie.lv inoculated with 0.3 x 10' M04 cells on day 0 On day 5 and 12. these mke were imsapentonemly in jet. ted with anti-(1)27 rn Ab 1 O'· (30 pgv. on day 8 arid :1:.5,- additional 4dS#.-:ofC02?:i|(tM^h'f5O:'pg) and vaccinated with ahti-miDecSdSWWA 0: |ig), TumPc gjTwth· was: measoidd: wdtlx ealipers 2 times a week, ’ikesetts awe skoxee Midget·© 24. in a timber stud). huOD27-l g and WT control, ή mice per group. were subcutaneously inoculated oath i x 10' M04 cells <·η tby 0. On day v and 12. these once were vaccinated wish amlMnDec20>OYA (5 pig; pi in- the I Lli agonist Ρ-..sly K"-LC ClOqg:): intsapeThoneaJly. On days 6 and id. mice we;e inserted with a ml-CD 7.7 mAh 1F5 DO pat Tumor growth was measured with calipers 2 times a week as indicated in the protocol illustrated m Figaro 285Λ. The results, shown in Figures 28 B. O and O. show the el tea of.no treatment {Figure 2 SB). vaccine treat mem alone (Rgurc 280) or saeetoe treatment in comhinmion with am; CD27 treatment {.Figure 281)1 on tumor ore in the πίκα as a tunction el the number ofdavs post turner inoculation. The mobm demonstrate that the cmnh;nation i rent mem o hi; an;b-('Tw7 mAh t iFD sign if;candy prolonged survival of tumor challenged mice:, ϊ^μμΜΜ 1 ¥5 Exhibits Potent Anti-Tumor .Activity In a Syngende Transgenic Moose rumor Challenge Model of the B€E$ B-hmphoma

To assess m viw anil-tumor activity of I F5. groups oi 0-10 hCD27 transgenic once ;Bnib/c hack.ground 1 were challenged with 10' BCLi B-lymphoma cells ndrni.niste.md intravenously on Day 0, Animals were then treated with 5 doses of ami-human CD27 mAh 1F5 as indicated. As shown in Figures 2bA and 2bB. significantly prolonged survival of lumot challenged mine, m a dose depe.nde.rn manner.

Tatmor Elllmg In Raji Xenograph SCO) Mouse Model CK.! ? SCID mice (purchased from Tavomc) were maintained in a pathogen-tree mouse tavtiity. Lymphoma Raji cells (1 x ! O'"Wen* iiuhcutaoeoualy injected into SCID mice. 4 mice per group. On day 6. these mii’^wcre treated with CD27 human m \hs via intrapemoncal administration. 0.5 mg per dose and dosed twice a week lor » weeks. Tumor growth was measured with calipew; a tunes a week. Results <4' tumor growth and Kaplun-Meier analysis are shewn in Figure X)A and 30B. iron· which is can be seen dan the awn CD27 rnAhs sisniileandy prolonged survival of the tumor challenged mice.

Id a farthm expoaritem. C8.17 SCID mict· tpurchased from Tueomc) were ntah dan ted in a palhogen-tVec mouse tat silty. Human lymphoma Rail cells p %. ItF) were subcuiarntfusiy injected Into Si ’ID mice on day 0. 0 mice per group. On day 5. these mice were treated with arfti~CD27 mAh IF5 eta mtmperitoneal-administration, 0-033.. 0.1 or 0.3 trig per dose and dosed twice a week her 3 weeks. Tut nor growth, was measured with caliper-' 7 time's a week.

The results, show n hi Figure 31 A. indicate that the antnCX.07 mAh {I i;5> signifunmily inhibited tumor growth and thus significantly prolonged survival of the tumor challenged mice. A Kaplan-Meier survival plot oi die data is also provided in Figure 31B, which chows that median survival eras increased by at least 10 days m mice from the treated group compared m the eatitml: group. !G5 and results .are shown id Figure 32, R cample 20

Tumor Killing in Pm&amp;H Xenograft SCID Moose Model (T<. 17 Si'll.) mice Ipiuehased from Taennii '> were maintained in a pathogen -irw ntousiv tueilnx, Human Kmj'homn I'amdi eel A ; I x H)') were subcutaneously injected Into St ID mice on day A (> msec per :ooup On she 5, these nn>.e were treated with ami· i'i.)27 hum-m mAh 1 FA , ia unm-'i-ruoneni adminisuaiiou. 0.033, 0-, 1 or 0,3 mg pet dose and dosed tx, o.e a week for 3 weeks To mm giowth 'was measured with calipers 2 limes a week.

The results shown in Figure 33. hi, Ik an* that the aiowC'IT;? mAh {1F3) gignilkantiy inhibited tumor gmxth t Figure 33-V> and thus significantly prolonged survival of the tiur-or challenged mice Kapl.m -Meier pim hi name MBe

Example 21

Anti-CD27 «*AI> Engineered ίο Not Bind Fc Receptors 1)0(¾ Not EaHanceT.Ceti Responses to a Ynedne Antigen

Ih.;CD27 transgenic mice were smmunmed vdth 5ua {vcd oi APO-mrgmed vueeme comprising ;m anb-mouse DH'Ou5 isO antibody Fused m ovaibumm {OVA} f re i erred tcs as mmObO-cO>OVA ?. in combination ft uh the·.' amPCDc? human tnAb HA i{.μ. ? os mAh 1 Fi mutant i l-c portion snu rated to prevent' Fc reccpmr binding s or eosnot igO mAh, One week Uftm\ splenoevir."·. were as tel y/ed for 008< Ί cell rwwuvstx to she OVA SiLNFEKL peptide (OVA peptide 277- 264} by-lF^'EtlSKlT·«$ getter ally described^

Tin." re oi it··:, shown in P· gt.no dm tk-monmtak' that the ahemd human w.AF \ FA dues noi enhastee the Twed responses to the \ nccs.no, and \ho<; non id be an effective a mem tor Nocking ihe CD7G/CD27 pathway jEonlydldots

Those skilled i d tbs cart wit! recogtaae, ostbe able to ascertain «aing no more th,an routine experimentation, many ecjoivalems of Oe specific embodiment* of the unoniio» described herein. Such eqni vuienia are intended to be encompassed by the following chums.

SUMMARY OF SEQUENCE LISTING

Claims (102)

1, An isolated human or humanized monoclonal antibody that hinds to 'huinan CD27, wherein the antibody exhibits at least one of the following properties: fa) blocks binding of sCD70 to CD27 by at least about 70% at an antibody concentration of 10 pg/ml; (h) binds to human CD27 with an equilibrium dissociation constant Md of l,0‘v M or less, or alternatively, an equilibrium association constant K.a of 10^ M"5 or greater; (c) induces specific complement mediated cytotoxicity (CDC) of €D27 expressingeells of at least 10% at an antibody coneenirarion of 3 ug/mi and approximately 6% rabbit serum complement; (d) induces antibody dependent cell-mediated cytotoxicity (ADCC) specific lysis of CD27 expressing cells of at least 10% at an; antibody concentration of 3 pg/nil and ratio of effector: target cells of 75:1 ; fe) improves median survival by at least 20%; in severe combined immunodeficiency (SCID) mice post tumor eel! inoculation in vim (5 x 10' Raji ceils or 1 x l(f Daudi cells) when administered at 0,3mg '(ip.) at least twice a week for 3 weeks compared fo mice to which antibody is not administered; (f) induces or enhances antigen-specific immune responses in combination with a vaccine or endogenous, antigen: (g) induces or enhances antigen-specific Till immune responses in combination with a vaccine or endogenous antigen; (h) induces or enhances anti geo-spec! fie T-cell proliferation or activation in combination with a vaccine or endogenous antigen: (i) reduces or inhibits T~cell proliferation or activation; (j) induces or enhances Ύ-cell activity when combined with simultaneous, separate or sequential TCR activation; (k) blocks binding of s€D?0 to CD27 by at least about 70% at an antibody concentration of 10 pg/ml and reduces or inhibits T-cel! activity when not capable of binding to, or having reduced binding to Fe receptors; (!) resells iii less than 50% depletion of CB3+ T-cells (oilier than NK ceils) in macaques when administered at 3.mg/kg (i.vi) over the period of 29 days immediately foliwing administration; or (m) results in less than 50% depletion of memory B~ce)ls In macaques when administered at 3 mg/kg (ire.) over the period of 29 days immediately following ad mini mat km. "2, An isolated monoclonal antibody that binds to 'human CID27 and induces or enhances an immune response against an antigen.

3, The antibody of claim 2, which induces or enhances a THI immune response.

4, An isolated monoclonal antibody that hinds to human CD27 and induces or enhances an immune response against an antigen, wherein the antibody induces or enhances T-eeil proliferation or acti vation,

5, Hie antibody of claim 4, whetein the antibody induces of enhances C13S+ T-eeil proliferation or activation,

6, The antibody of claim. 4 or 5, wherein the antibody further inhibits the binding of CD70 to CD27 on cells.

7, The antibody of any one of claims 2-6., wherein the antibody comprises SEQ ID NOs: 37 and/or 43,

8,. The antibody of any one of claims 2-5, wherein the antibody binds to human CD27 and does not significantly inhibit the binding of CD70 to CD27 on cells.

9, The antibody of claim 8, wherein the antibody comprises SEQ ID NOs: 7, 13, and/or 19.

10. The antibody of any non of claims 2-9, wherein the antibody comprises an Fc region capable of effective binding to Fc receptors,

11. A bispecific molecule comprising the antibody of arty one of claims 210 linked to a second molecule having a binding specificity which;is..different from the antibody.

12. The bispeeriie molecule of claim 11, wherein the second molecule binds to a T cell receptor, .! 3, The bispecific molecule of claim 12, wherein- the T cell receptor is selected from the group consisting of CD3, €134(5 and €D23

14, The btspeei.fic molecule of claim 11, wherein "the second molecule binds to an Fc receptor.

15. The bispecific molecule of claim. 1.1, wherein the second molecule binds to an N K .receptor,

16, An isolated monoclonal antibody that hinds to human. €1327 and inhibits the binding ofCD7G to CD2? on cells, wherein the antibody inhibits binding of s€D70 to CD27 expressing, cells by at least about 70% at an antibod y concentration of 10 pg/ml and inhibits or reduces an immune response to an anti gen.

17. The antibody of claim 16, which Inhibits or reduces T-eell proliferation. or activation.

8. The antibody of claim, 1..6 or 17, which inhibits or reduces CDS-r T-cel! proliferation or activation, 1.9, Tlw antibody of any one of claims 16-18, wherein the antibody comprises SEQ ID NOs: 37, 43,49, 55, 103, and/or 1.09,

20. The anybody of any one of claims 16-1.8, wherein the antibody comprises SEQ ID NOs: 37 and/or 43.

21. The antibody of any one of claims 16-20, whembt..the-aptibody N a fragment without a functional Fc domain.

22. The antibody of any one of claims 16-20, wherein the antibody includes a mutated Fe region displaying reduced or no binding to 1¾ receptors·,

23. An isolated monoclonal antibody that binds to human CD27 and Induces or enhances effector ceil function, wherein the antibody induces at least about 40% ADCC specific lysis of CD27 expressing-ceils at a concentration of at an antibody concentration of 1.0 μ g/m! or Induces at least about 40% complement mediated cytotoxic Uv (CDC) of CD27 expressing cells at a concentration of 10 pg/nil

24. The antibody of claim. 23, wherein, the .antibody comprises SEQ ID NOs; 61, 67, 85,91, 97, 37, 43,49, 53, 19,49,35, 103, and/or 109,

25. Hie antibody of any one of claims 23-24, wherein the antibody inhibits the binding of CD7Q to GD27 on cells,

26. The antibody of claim 25, wherein.the antibody comprises SEQ ID NOs: 37,43, 49, 55, 19, 103, and/or 109.

27. The antibody of claim 23 or 24, wherein the antibody does not inhibit the binding of CD70 to CD27 on cells,

28. The antibody of claim, 27, wherein the antibody comprises SEQ ID NOs; 61, 67, 85, 91, 97, 7, 13, and/or 19. 2,9, The antibody of claim 28, wherein the antibody comprises SEQ ID NOs: 37,43,7,13, and/or 19.

30, The antibody of any non of claims 23 -29, wherein the antibody is a foil length antibody.

31, The antibody of any one of the preceding claims, wherein the antibody comprises a second binding specificity for an pe receptor.

32, The antibody of any one of the preceding claims, wherein the CD27 expressing.cells are B cells.

33, The antibody of any one- of claims .1--31, wherein the CD27 expressing cells are T cells.

34, The antibody of any one of claims 1-31» wherein the CD27 expressing cells are OD8+T.cells,

35, The antibody of any one of the preceding claims, wherein the CD27 expressing cells are selected from the group consisting of iurkat cells, Raji ceils, Ramos cells and Baud! Cells.

36, The antibody of any one of the preceding claims, wherein· the CD27 expresslng cells· are tumor cells,

37, The antibody of any one of the preceding claims, wherein the antibody Is selected from the group consisting of an IgGTan lgG2, an IgG3* an lgG4,an IgM, an IgA I, an IgA2,an IgD, and an IgE antibody,

38, An isolated monoclonal antibody, wherein the antibody binds to, human CD27 and comprises a heavy chain variable region derived from a human gerailine. Vn 3*7 or Vh 3-33 gene, 39» An isolated monoclonal antibody, wherein the antibody binds to human CD27 and comprises a light chain variable region derived from, a human germline VT3-30, VK3~!1, Yk IB-lb, or VK 1-13 gene.

40. An isolated monoclonal antibody, wherein the antibody binds to human. CD27 and comprise?; a heavy- chain variable region derived from a human germiloe Vh 3-7 or ¥h .3-33 gene and a light chain variable regiqfrrierivedfroM a human germ line VK 3 20, Yk 3-11, VK ID-16, or YR 1 -13 gene,

41. An isolated: monoclonal an d body w hich binds to h uroan CD2?, wherein the antibody comprises a heavy chain variable region comprising CD-R!. ('DR2, and €DR3 sequences and a light chain variable region comprising CORE C.DR2, and CDR3 sequences, wherein, the heavy drain variable region CDR3 sequence comprises an amino acid sequence selected, from the consensus sequence; R 1(1,1-..0) (S,L,G,-) (G,L,T,W? -) ίΝ,Α,Τ,ΙΙ· ; (V.T.~) AIR·} (G.V.· ) (R, --} (G,M, -) CD,HXJVW) (A,G,N,W) (D,F,Y,Y) (F,L) t D,E) (Η,Ι,Ι,,Υ) (SEQ ID NO; 113), wherein “-“ denotes the option of no amino acid residue being present at that consensus position,

42. The antibody of claim.41-, wherein, the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the consensus sequence: Q (F.R,Y) iN.S) N.T,S) (Y,W> P (F,L,P,R) T (SEQ ID MO: 114), wherein denotes the option of no amino acid residue being present at that consensus position.

43. The antibody of claim 42, wherein the heavy chain, variable region CDR2 sequence comprises an amino acid sequence selected from the consensus sequence: 1 (K,W) <Y,N,Q) D G S (EM <KSQ) (SEQ ID MO: 11.5), wherein “-“ denotes the option of no amino acid residue being present at that consensus position, and the light chain variable region CDR2 sequence comprises an amino acid sequence selected from, the consensus sequence: (AJD.i A S (SEQ ID HO: 116),

44. The antibody of claim 43, wherein, the heavy chain variable region CDRI sequence comprises an. amino acid sequence selected from the consensus sapience: G P (T,S) (FX) (S,N) G,S,H) (Y,H) (SEQ ID NO: 117); and the light chain variable region CDRI -sequence comprises an amino add sequence selected from the consensus sequence: Q (DiXS) (i;V) (D,S) (R,S) (A,W,Y) (SEQ ID NO; 118).

45. An iso f ated monodon al antibody which binds to human CD27 and comprises heavy and Light chain variable region CDR1, CDR2 and:CDR'3 sequences.selected from the group co.n.si.sd.ng oh (i) a heavy chain variable region CDRI comprising an amino acid sequence selected tern the consensus sequence: G F (T,S) (F,L·) (S,H) {I,S»H} (Y,B) (SEQ ID NO: 117); (ii) a heavy chain variable region 'CDR2 comprising an amino add sequence selected from the .consensus sequence: Ϊ (K,W) (Y»N,Q) DG S ·Π.,Χ > (K,Q> (SEQ ID .NO: 115); (lii) a heavy chain variable region CDR3 comprising an amino add sequence selected from the consensus sequence: R ΐ< ί J J>> (8,1,,0,-) (G,E,T,W, -) (RAXH,~) IV.T.-} AIR-} (G.V.- ) (R, -} (CLM, -) CD,H,LJVW) (A,G,M,W) (PJ-.V..Y) iPJ mH,E) (HJ.LY) (SEQ ID NO 113); (iv) a light chain, variable region CDRI comprising an amino acid sequence selected from the consensus sequence: Q (D,G,5) (!,¥) (D,S) (R,S) (A,W,Y) (SEQ ID NO: 118); iv) a light chain variable region CDR2 comprising an amino aeid sequence selected Item the consensus sequence: (A,D) A S (SEQ ID NO: 1.16); (v-i) a light chain variable region CDR3 comprising an amino acid sequence selected from the consensus sequence; Q (F,R,Y) (N,S) (N,TJ) (Y,W) P (F,L,P,R) T (SEQ: ID MO: 114); wherein.'denotes the option of no amino acid residue being present er that consensus position.

46. An isolated monoclonal, antibody which 'hinds to .human. GD27, wherein the antibody comprises heavy and light chain, variable regions comprising CDRI, CDR2, and CDR3 sequences, wherein, (a) the heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ I'D NOs: 10, 28, 40, 52, 64, 76, SB, 106, and conservative amino acid sequence modifications thereof; and (b) the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1.6, 34. 46,58, 70, 82,94, 100,112, and conservative amino acid sequence modifications thereof.

47, The antibody of claim 46, wherein the heavy chain variable region CDR2 sequence comprises: an amino acid sequence selected from the group consisting of SEQ ID NOs; 9, 27, 39, 51,63, 75, 87, 105, and conservative amino add sequence modifications thereof,.and the 'light.chain- variable region CDR2 sequence comprises an amino add sequence selected from the group consisting of SEQ ID NOs: 15, 21,33, 45, 57, 69, 81,93, 99, 111, and conservative ammo acid sequence .modifications thereof.

48, The antibody of claim 47, wherein the heavy chain variable region CDRi sequence comprises ao. amino acid sequence selected from the group consisting of SEQ ID NOs: 8,26, 38, 50,62,74, 86, 104, .and conservative amino acid sequence modifications thereof; and the lightchain variable region CDRI sequence is selected from the group consisting of 8EQ ID NOs: 14, 20, 32,44, 56, 68,.80, 92, 98,110, and conservative amino add. sequence modifications thereof

49, An isolated monoclonal antibody which hinds to human CD27, wherein the antibody comprises; (I) a heavy chain variable region CDR1 comprising SEQ ID NO; 38; a heavy chain, variable region CDR2 comprising SEQ ID NO; 39; a heavy chain variable region CDR3 comprising SEQ ID NO: 40: a light chain; variable region CDRI comprising SEQ IP NO: 44; a light chain variable region CDR2 comprising SEQ ID NO: 45; and a light Cham variahle region. CDR3 comprising SEQ ID. NO: 46; fit) a heavy chain variable region. GDRI comprising SEQ ID NO; 50; a heavy chain variable region CDK2 comprising SEQ ID NO; 51; a heavy chain variable region COR3 comprising SEQ ID NO:. 52; a light chain, variable region CDR I comprising SEQ ID NO: 56; a light chain variable region CDR2 comprising SEQ ID NO: 57: and a light chain variable region CDR3 comprising SEQ ID NO: 58; (ill) a heavy chain variabfe region CDRI comprising, SEQ ID NO: .104: a heavy chain variable region CDR2- comprising SEQ ID NO: i 05: a heavy chain variable region CDE3 comprising SEQ ID NO: 106; alight chain variable region GDR1 comprising SEQ ID NO: 110; a light chain variable region CDR2 comprising' SEQ ID NO: 111; and a light chain variable region CDR3 comprising SEQ ID NO: 112; ilv) a heavy chain variable region CDRI comprising SEQ ID NO: 86; a heavy chain variable region CDR2 comprising SEQ ID NO: 87; a heavy chain variable region CD R3 comprising SEQ ID NO: 88: a light chain, variable .region CDRI comprising SEQ ID NO: 92 or 98; a .light chain variable region CDR2 comprising SEQ ID NO: 93 or 99; ancl a light chain variable region CDR3 comprising SEQ ID NO: 94 or 100; iv) a heavy chain, variable region CDR1 comprising SEQ ID NO: 26; a heavy chain variable- region CDR2 comprising SEQ ID NQ: 27.; a. heavy chain variable region CDR3 comprising SEQ ID NO: 28; a light chain variable region CDR! comprising SEQ ID NO: 32; a light chain variable region CDR2 comprising SEQ: ID NO: 33; and a light chain variable region CDR3 comprising SEQ ID NO: 34; (vi) a heavy chain variable region CDR.!. comprising SEQ ID NO: 74; a heavy chain variable·,region CDR2 comprising SEQ ID NO: 75; a heavy chain variable region GDR3 comprising SEQ ID NO: 76; a light chain variable region CDRI comprising SEQ ID NO: 80; a light chain variable region CDR.2 comprising SEQ ID NO: 81; and alight chain .variable region CDR3 comprising SEQ ID NO: 82: (vii) a heavy chain variable region CDR1 comprising SEQ ID NO: 8; a heavy chain variable region CDR2 comprising SEQ ID NO; 9; a heavy chain variable region CDR3 comprising SEQ ID NO: 10; a light chain variable .region CDRI ecmiprisitig SEQ ID NO: 14 or 20; a light chain variable region CDR2 comprising SEQ ID NO: 15 or 21: and a light chain variable region GDR3 comprising SEQ ID NO: .16 or 22; or (viii) a heavy chain variable region CDRI comprising'SEQ IP NO: 62; a heavy chain variable region CDR2 comprising SEQ ID NO: 63; a heavy chain variable region CDR3 comprising SEQ ID NO: 64; a light chain variable region CDRI. comprising'SEQ'ID NO: 68; a light chain, variable region CDR2 comprising SEQ ID NO; 69; and a light chain variable region, CPB3 comprising SEQ ID NO: 70,

50, An isolated monoclonal antibody which binds to human CD27 and comprises a heavy chain variable region comprising an amino acid sequence which is. at least 90% identical to the amino acid sequence selected, from the group consisting of SEQ ID NOs: 6, 7, 24, 25. 36, 37,4% 49, 60, 61,72,73,84, 85,102, and 1.03.

51, An isolated, monoclonal antibody which binds to human CD27 and comprises a light chain variable region, comprising an amino acid: sequence which is at least. 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NOs: 12, .1.3, 18,19, 30,31,42, 43,54,55,66,67, 78, 79,90, 91, 96, 97,108, and 109.

52, An isolated, monoclonal antibody which binds to human CD27 and comprises a heavy chain variable region and a light chain variable region comprising an amino acid sequence ai least 90% Identical to the. amino acid sequences selected from, the group consisting of: (a) SEQ ID NOs; 37 and 43, respectively; (b) SEQ ID NOs: 49 and 55, respectively; (e) SEQ ID NOs: 1.03 and 109, respectively; id) SEQ ID NOs: 85 and 91, respectively; (e; SEQ ID NOs: 85 and 97, respectively; fi) SEQ ID NOs; 25 and: 31, respectively: (g) SEQ ID NOs: 73 and 79, respectively: (h) SEQ ID NOs: 7 and 13, respectively; (it SEQ ID NOs; 7 and Irrespectively; and (j) SF.Q ID NOs: 61 and 67, respectively,

53. An isolated monoclonal antibody, wherein the antibody binds to human CD27 and comprises: (μ) a heavy chain variable region comprising SEQ ID NO:37, or conservati ve amino acid substitutions thereof, and a light chain, 'variable region comprising SEQ ID NO; 43, or conservative amino acid substitutions thereof; (b) a heavy chain variable region comprising SEQ IP NO: 49. or conservati ve amino acid substitutions thereof, and a light chain variable region comprising SEQ ID NO: 55, or conservative amino acid substitutions thereof; (c) a heavy chain variable region comprising SEQ ID NO; 103, or conservative amino acid substitutions thereof, and a light chain variable region comprising SEQ ID NO: 109, or conservative amino acid substitutions thereof; (d) &amp; heavy chaift variable region comprising SEQ: ID NO: 85, or conservative amino acid, substitutions thereof, and a light chain variable region comprising SEQ ID NO: 94, or conservative amino add substitutions thereof: (e) a heavy chain variable region comprising SEQ ID NO: 85, or conservative amino acid substitutions thereof, and a light chain variable region comprising SEQ ID NO: 97, or conservative amino acid substitutions thereof; (I) a heavy chain variable region comprising SEQ ID NO; 25, or conservative amino acid substitutions thereof, and a light chain variable region comprising SEQ ID NO: 31, or conservative-amino n<. id substitutions thereof; (g) a heavy chain, variable region comprising SEQ ID NO: 73, or conservative amino acid: snbstttn lions thereof, and a light chain variable region. comprising SEQ ID NO: 79, or conservative amino.acid substitutions thereof; (h) a heavy chain variable region comprising SEQ I'D- NO; 7» or conservative amino acid substitutions thereof, and a light chain, variable region comprising SEQ ID NO: 13, or conservative amino acid substitutions .thereof; (t) a heavy chain variable region comprising SEQ ID NO: 7, or conservative amino acid substitutions thereof, and a light chain, 'variable region comprising SEQ ID NO; 19, or conservative amino acid substitutions thereof; or (j) a heavy chain variable region comprising SEQ ID NO: 61, or conservative amino acid substitutions·thereof and. a light chain variable region comprising SEQ ID NO: 67, or conservative amino acid substitutions thereof

54. Ail Isolated antibody that competes for binding to human 0327 with the antibody of claim 52,

55. An isolated, antibody which binds to an epitope bound by the antibody of claim 52 ,

56. The antibody of any one of the preceding claims, wherein the antibody Is a human or humanized, antibody.

57. An isolated antibody that binds to human- CD27 and'competes for binding with an antibody comprising heavy and light chain variable regions'comprising: (a) SEQ I'D NOs; 37 and 43, respectively; (b) SEQ ID NOs: 49 and 55, respectively; (e) SEQ ID NOs; 103 and 109, respectively; id) SEQ ID NOs: 85 and 91, respectively; (e) SEQ ID NOs; 85 and 97, respectivel y; (1) SEQ ID NOs: 25 and. 31, respectively; (g) SEQ ID NOs: 73 and 79, respectively; (h) SEQ ID NOs: 7 and 13, respectively; (i) SEQ ID NOs; 7 arid 19, respectively; or φ SEQ ID NOs; 61 md 67, respectively.

58. An isolated antibody that binds to an epitope on human CD27 bound fey an antibody comprising heavy and light chain variable regions comprising; (a) SEQ: ID NOs; 37 and 43, respectively; (h) SEQ: ID NOs: 49 and 55, respectively; (c) SEQ ID NOs: 103 and 109, respectively; (d) SEQ ID NOs; 85 and 91, respectively.; (e) SEQ ID NOs; 85 and 97, respectively; (1) SEQ ID NOs: 25 and 31, respectively; (g) SEQ ID NOs; 73 and 79, respectively; (h) SEQ ID NOs; 7 and 13, respectively; (i) SEQ ID NOs; 7 and 19, respectively; or if) SEQ ID NOs: 61 and 67, respectively, .59. An isolated monoclonal antibody that binds human CD27, wherein the antibody comprises, a heavy chain variable region and a light chain variable region encoded by nucleic acid sequences selected from the group consisting oE (a) SEQ ID NOs: 5 and 11, .respectively; (b) SEQ ID NOs; 5 and 17, respectively; (c) SEQ ID NOs; 23 and 29, respectively; (d) SEQ ID NOs: 35 and 41, respective,ly: (e) SEQ ID NOs: 47 and 53, .respectively: If) SEQ ID NOs: 59 and 65, respectively: Cg) SEQ IP NOs; 71 and 77, respectively; (h) SEQ ID NOs: 83 and 89,. (I) SEQ ID NOs; 83 and 95; and (j) SEQ ID NOs: 101 and 107, respectively or nucleic acid sequences having at least 909¾ identity to the nucleic acid sequences of (a)- Φ>

60. An expression vector comprising a nucleotide sequence encoding the variable region of a light chain, heavy chain, or both light and hea vy chains of an antibody which binds Jinman CD27 as claimed in claim 50 or 51.

61. A cell '.transformed with an. expression vector of claim 60,

62, The antibody of any one of claims 1-10 or 16-59, wherein the antibody is selected from the group consisting of an IgGL an IgG2, an lgG3, an IgG4, an IgM, an IgAl, an IgA2, an IgAsec, an IgD, and: an IgE antibody.

63, The antibody of any one of claims 1-10 or 16-59 wherein the antibody is a hums», humanized or chimeric antibody.

64, A corn.positiQn eomprislng the antibody or hlspeellle molecule of any one of claims 1 -59 and a carrier,

65. The composition .of claim 64, further comprising an adjuvant,

66, Hie composition of claim 64 or 65, further comprising an immnnstimulafcry agent,

67. The composition of claim 66, wherein the imm.unostimulatory agent is selected from the group consisting of CD40 ligand, FLT 3 ligand, cytokines, colony-stimulating factors, an anti-GTLA-4 antibody, LPS (endotoxin), ssRNA, dsRNA, BaciOe Calmeite-Goerin (BCG), Imvamisole hydrochloride, intravenous, immune globulins and a Toll-like Receptor (TLR) agonist,

68. The composition of claim: 67, wherein the Toll-like Receptor agonist is selected from the group consisting of a TLR 3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, and a TLR 9 agonist,

69, The composition of claim 64, further comprising an immunosuppressive agent.

70. The composition of claim 64, further comprising another antibody.

71. The composition of claim 64 further comprislug m. antigen.

72. The composition of claim 71, wherein the antigen, comprises a component of a pathogen,

73. The composition of claim 71, >vhemi« the antigen comprises a tumor antigen, allergen or an autoaiitlgen,

74. The composition of claim. 71, wherein the antigen comprises a tnmor antigen,

73, The composition of claim 74, wherein the tumor antigen is selected from the group consisting of phCG. gplOO or Pmell7, RBR2/nem WTL mesothelm, QBA, gp i00, MARTI, TRP-2. meian-Λ, NY'-USO-I. NY-RR-1, N Y-CO-58, MN (gp250), icflotype, M&amp;OE-l,MAGEv3,MAGE-A3., Tyrosinase, Telomefase, SSX2 antigens, MIKTT antigens, and genu cell derived, tumor antigens,

76, A method, for inducing or enhancing an immune response against an. antigen in a subject comprising administering to the subject an antibody, composition or hispecific molecule of any one of the preceding claims in an amount effective to.induce or enhance an. immune response against an antigen,

77, The method of claim 77, wherein the antibody Inhibits the binding of CD70. to CD27 on cells,

78, The method, of claim 77, wherein the antibody comprises SEQ ID NOs: 37 and/or 43,

79- The method of claim 76, wherein the antibody does not inhibit the binding of C2D70 to C.D27 on cells.

80, The method of claim 79, wherein the antibody comprises SEQ ID NOs: 7, 13, and/or 19.

81, The method of any one of claims 76-80, *\ ir In the antibody is a Ml length antibody,

82, A method of inhibiting growth, of CD27 expressing cells comprising contacting the cells with, an antibody Or composition of any one-of the preceding .claims. In an amount effective to inhibit growth of €D27 expressing cells.

83, The method of claim 82, wherein the cells are contacted with an antibody in the presence of effector cells under conditions sufficient to induce antibody-dependent cellular cytoxicity (A’DCC) of target cells.

84, The method of claim 83, wherein the antibody induces at least about 4078 ADCC mediated specific lysis of CD27 expressing cells at a concentration of i 0 pg/ml and comprises SEQ ID. NOs: 61, 67, 85, 91, 97, 37, and/or 43.

85, Hie method of claim 82, wherein the cells are contacted, with an antibody under conditions sufficient to induce CDC of said cells,

86- The method of claim. 85, wherein the antibody induces at least about 40% CDC Of CD27 expressing ceils at a concentration of 10 pg/rnl and comprises SEQ ID NOs: 7, .13, 19,49,55, 103, and/or 109.

87. The method of claim 82, wherein the antibody binds to human €027' and blocks or prevents the binding of CD70 to CD27 on cells,

88. The method of claim 87, wherein the antibody' comprises SEQ ID NOs: 37,43, 7, 13, 1.9, 103, and/or 109,

89. The method of churn 82, wherein the antibod y binds to h uman CD27 and does not block or pre vent the binding of CD70 to CD27 on cells.

90 , The·' method of claim 89, wherein the antibody comprises SEQ ID NOs: 61,67, 85., 91,97, 7,13, and/or .19,

91, The method of .claim 83, wherein the antibody comprises SEQ ID MOs: 37,43, 7,13, and/or 19.

92, The method of any one of claims '82-91, wherein the: antibody- is a Ml. length antibody,

93, The method of of any one of claims 82-91, wherein the antibody contains a second binding specificity for an. Fe receptor,

94, The method of of any one of claims 82-9.1, wherein the cells are tumor cells.

95, The .method of claim. 82, wherein the disorder' Is a cancer selected from the group consisting of leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, myeloblasts promyelocyte myelomonoeyric monocytic erythroleukemia, chronic leukemia, chronic myelocytic (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkltr s lymphoma and marginal zoneB cell lymphoma. Polycythemia vera Lymphoma, Hodgkin's disease, non-Hodgkin* s disease, multiple myeloma, Waldenstrom's macroglobulmerma, heavy chain disease, solid tumors, sarcomas,-and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chroadrosareoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarooma, lymph angiosarcoma, lyniphangioendothehosareoraa, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon sarcoma, colorectal carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, sc]uamous cel! carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma* papillary carcinoma, papillary adenocarcinomas, cystadenoeaminomm medullary carcinoma, bronchogenic carcinoma, renal cel! carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilni's tumor, cervical, cancer, uterine cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, roc small cel! lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pioeaforna, hemangioblastoma, acoustic neuroma, oligodendroglioma, roenangtoma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal, carcinoma, esophageal carcinoma, basal cell carcinoma, biliary tract cancer, bladder cancerybone cancer, brain and central nervons system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancers, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer, intraepithelial neoplasm, kidney cancer, larynx cancer, liver cancer, lung cancer (small cell, large cell), melanoma, neuroblastoma; oral cavity cancer(for example lip, tongue, mouth and pharynx), ovarian cancer, pancreatic cancer, retinoblastoma, idiabdomyosarcoma, rectal cancer; cancer of the.respiratory system,.'sarcoma, skin, cancer, stomach, cancer, testicular cancer, thyroid cancer, uterine cancer, and cancer of the urinary system,

96. The method of claim 82, wherein the disorder is a cancer selected from die group consisting of chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lyrnphoma, Burkitt’s lymphoma and marginal zone B ceil lymphoma.

97. The method of claim 82, wherein the disorder is selected from the group of disorders consisting of bacterial, fungal, viral and parasitic infectious diseases.

98. A method inhibiting the binding of CD7C) to CD27 on cells in a subject having a disorder by administering to the wb;ect the antibody, bispeeific molecule, or composition, of any one of the preceding claims

99. A method of down-regulating a T cel ! 'response in an indi vidual having a disorder by administering to a subject the antibody, bispeeific molecule, or composition according to any one of the preceding claims,

100. The method of claim 98, wherein the subject is administered, an antibody comprising SEQ ID NOs; 37,43,49, 55, 103, and/or 109. 10!, The method of claim 1.00, wherein the antibody comprises SEQ ID NOs: 37 and/or 43.

102. The method of any one of claims 98-101, wherein, the antibody is a Fab fragment.

103. The method of any one of claims 98-10 !, wherein the antibody comprises a mutated Fc region.

104. The method, of arty one of claims 98 -101, wherein the disorder is selected from the group consisting of graft rejection, allergy and an autoimmune disease.

105. The method Of claim 104, wherein the autoimmune disease is selected from the group consisting of multiple' sclerosis, rheumatoid arthritis, type 1 diabetes, psoriasis, Crohn’s disease and. other inflammatory bowel diseases such as ulcerative colitis, systemic lupus eyfhemaCosns (SIT*), autoimmune encephalomyelitis, myasthenia gravis (MCI), Hashiraoto’s thyroiditis, Goodpasture's syndrome, pemphigus. Graves disease, autoimmune hemolytic anemia, antoimmnne thrombocytopenic purpura, scleroderma with anti- collagen antibodies, surg'd connective tissue disease, polypyositls, pernicious anemia, idiopathic Addison’s disease, autoimmune associated infertility, glomerulonephritis, crescentic glomerulonephritis, proliferative glomerulonephritis, bullous pemphigoid, Sjogren's .syndrome, psoriatic arthritis, insulin resistance, autoimmune diabetes meilitus, autoimmune hepatitis, autoimmune hemophilia, .autoimmune lymphoproliferative syndrome (ALPS), aetoirnmune hepatitis, autoimmune hemophilia, autoimmune lympimprolifemtive syndrome, autoimmune uveorebmtis, Gulilarn-Baxe syndrome, axterioseleiwsis and Alaheiroer’s disease.

106. A method fox detecting the presence or absence of CD27 in a .biological sample, comprising: (a) contacting .a biological sample with an antibody of any one of claims '1-10 ox 16-59, wherein the antibody is labeled, with a detectable substance: and (b) detecting the antibody bound to GD27 to thereby detect the presence or absentee of CD27 in the biological sample. 107, A. method for enhancing ail immune response against an antigen in a subject in need thereof % administering to the subject: I) a. human orImmunized anti-CD.27 antibody and ii) the antigen, wherein the anu--CD27 antibody is administered separately from and before the antigen is ..administered, 1 OS, A method as claimed in claim 107 wherein the anti~€D27 antibody is administered at least 2 hours before the antigen,

109, A method as claimed in el aim, 107' wherein the a.nti-CD27 antibody is administered at least 12 hours before the antigen,

110, A method as claimed in claim 107 wherein the anb--€D2? antibody is administered at least 24 hours before the antigen.

111, A method as claimed in claim 107 wherein the aoti~CP27 antibody is administered at least 48 hours before the antigen.

112, A method as claimed. In claim, ,107 wherein the anti-CD27 antibody is administered at least 72 hours before the antigen,

113, A method as claimed in claim 107 wherein the anti-€D27 antibody is administered: between·at least 2 and 96 hours before the antigen. 1.14. A method for enhancing an Immune response against an antigen in a subject in need thereof by slmultanously, separately or sequentially administering to the subject:' iya human or humanized ahti-.GD2? antibody; ii) a TLR agonist; and lit) optionally, the antigen, 115. A. method as cl aimed in claiml 14 wherei n the TLR, agonist is a TLR3 agonist, 1,16. A method as claimed in claim 114 wherein the TLR agonist is Poly 1C: LC.