WO2006025580A1 - 抗ヒストンh1モノクローナル抗体およびこれを産生するハイブリドーマ - Google Patents
抗ヒストンh1モノクローナル抗体およびこれを産生するハイブリドーマ Download PDFInfo
- Publication number
- WO2006025580A1 WO2006025580A1 PCT/JP2005/016268 JP2005016268W WO2006025580A1 WO 2006025580 A1 WO2006025580 A1 WO 2006025580A1 JP 2005016268 W JP2005016268 W JP 2005016268W WO 2006025580 A1 WO2006025580 A1 WO 2006025580A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- histone
- mammal
- monoclonal antibody
- transplant rejection
- Prior art date
Links
- 230000002583 anti-histone Effects 0.000 title claims abstract description 48
- 210000004408 hybridoma Anatomy 0.000 title claims abstract description 34
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 50
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 43
- 229920001184 polypeptide Polymers 0.000 claims abstract description 41
- 238000003745 diagnosis Methods 0.000 claims abstract description 7
- 108010033040 Histones Proteins 0.000 claims description 72
- 238000000034 method Methods 0.000 claims description 58
- 241000124008 Mammalia Species 0.000 claims description 56
- 239000000427 antigen Substances 0.000 claims description 48
- 108091007433 antigens Proteins 0.000 claims description 46
- 102000036639 antigens Human genes 0.000 claims description 46
- 206010052779 Transplant rejections Diseases 0.000 claims description 44
- 239000003018 immunosuppressive agent Substances 0.000 claims description 29
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 26
- 230000001506 immunosuppresive effect Effects 0.000 claims description 25
- 229940125721 immunosuppressive agent Drugs 0.000 claims description 15
- 229960003444 immunosuppressant agent Drugs 0.000 claims description 14
- 210000000170 cell membrane Anatomy 0.000 claims description 13
- 239000012472 biological sample Substances 0.000 claims description 12
- 230000001861 immunosuppressant effect Effects 0.000 claims description 10
- 230000036961 partial effect Effects 0.000 claims description 9
- 206010062016 Immunosuppression Diseases 0.000 claims description 8
- 239000004480 active ingredient Substances 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 102000006947 Histones Human genes 0.000 claims description 4
- 239000000032 diagnostic agent Substances 0.000 claims description 4
- 229940039227 diagnostic agent Drugs 0.000 claims description 4
- 210000004988 splenocyte Anatomy 0.000 claims description 4
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 229920000831 ionic polymer Polymers 0.000 claims 1
- 210000000056 organ Anatomy 0.000 abstract description 22
- 230000001900 immune effect Effects 0.000 abstract 1
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 41
- 239000002609 medium Substances 0.000 description 26
- 239000000243 solution Substances 0.000 description 23
- 238000002054 transplantation Methods 0.000 description 23
- 239000000872 buffer Substances 0.000 description 18
- 241000700159 Rattus Species 0.000 description 16
- 238000007799 mixed lymphocyte reaction assay Methods 0.000 description 16
- 210000004989 spleen cell Anatomy 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 210000004698 lymphocyte Anatomy 0.000 description 12
- 239000012228 culture supernatant Substances 0.000 description 11
- UZOVYGYOLBIAJR-UHFFFAOYSA-N 4-isocyanato-4'-methyldiphenylmethane Chemical compound C1=CC(C)=CC=C1CC1=CC=C(N=C=O)C=C1 UZOVYGYOLBIAJR-UHFFFAOYSA-N 0.000 description 10
- 230000007910 cell fusion Effects 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 238000001262 western blot Methods 0.000 description 9
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 8
- 206010035226 Plasma cell myeloma Diseases 0.000 description 8
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 201000000050 myeloid neoplasm Diseases 0.000 description 8
- 238000010186 staining Methods 0.000 description 8
- 239000000725 suspension Substances 0.000 description 8
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 8
- 210000004185 liver Anatomy 0.000 description 7
- 230000001235 sensitizing effect Effects 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 239000002202 Polyethylene glycol Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 239000006285 cell suspension Substances 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 229920001223 polyethylene glycol Polymers 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 229960001967 tacrolimus Drugs 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 230000000903 blocking effect Effects 0.000 description 5
- 210000002865 immune cell Anatomy 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 238000001114 immunoprecipitation Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 229960004857 mitomycin Drugs 0.000 description 4
- 238000007447 staining method Methods 0.000 description 4
- 230000004936 stimulating effect Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 239000002033 PVDF binder Substances 0.000 description 3
- 108010067902 Peptide Library Proteins 0.000 description 3
- 239000012979 RPMI medium Substances 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 239000012148 binding buffer Substances 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 210000002216 heart Anatomy 0.000 description 3
- 238000010255 intramuscular injection Methods 0.000 description 3
- 239000007927 intramuscular injection Substances 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004091 panning Methods 0.000 description 3
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000002062 proliferating effect Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 238000010254 subcutaneous injection Methods 0.000 description 3
- 239000007929 subcutaneous injection Substances 0.000 description 3
- 210000001541 thymus gland Anatomy 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 2
- 108010036949 Cyclosporine Proteins 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 108010047956 Nucleosomes Proteins 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 101710120037 Toxin CcdB Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229960002170 azathioprine Drugs 0.000 description 2
- -1 azathioprine Steroids Chemical class 0.000 description 2
- 229960004669 basiliximab Drugs 0.000 description 2
- 238000007413 biotinylation Methods 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229960001265 ciclosporin Drugs 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000013024 dilution buffer Substances 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 229960005150 glycerol Drugs 0.000 description 2
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 239000007951 isotonicity adjuster Substances 0.000 description 2
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 238000000691 measurement method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229920005615 natural polymer Polymers 0.000 description 2
- 210000001623 nucleosome Anatomy 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 238000003127 radioimmunoassay Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 210000003705 ribosome Anatomy 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 229920001059 synthetic polymer Polymers 0.000 description 2
- 239000002562 thickening agent Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 229930105110 Cyclosporin A Natural products 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 238000006852 Griffith reaction Methods 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- 108091036060 Linker DNA Proteins 0.000 description 1
- HZQDCMWJEBCWBR-UUOKFMHZSA-N Mizoribine Chemical compound OC1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 HZQDCMWJEBCWBR-UUOKFMHZSA-N 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000711408 Murine respirovirus Species 0.000 description 1
- 241000282520 Papio Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 239000006180 TBST buffer Substances 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 235000010724 Wisteria floribunda Nutrition 0.000 description 1
- BTKMJKKKZATLBU-UHFFFAOYSA-N [2-(1,3-benzothiazol-2-yl)-1,3-benzothiazol-6-yl] dihydrogen phosphate Chemical compound C1=CC=C2SC(C3=NC4=CC=C(C=C4S3)OP(O)(=O)O)=NC2=C1 BTKMJKKKZATLBU-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 230000000340 anti-metabolite Effects 0.000 description 1
- 229940100197 antimetabolite Drugs 0.000 description 1
- 239000002256 antimetabolite Substances 0.000 description 1
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 1
- LMEKQMALGUDUQG-UHFFFAOYSA-N azathioprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC=NC2=C1NC=N2 LMEKQMALGUDUQG-UHFFFAOYSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000006287 biotinylation Effects 0.000 description 1
- 210000002798 bone marrow cell Anatomy 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 230000009702 cancer cell proliferation Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229930182912 cyclosporin Natural products 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 102000054766 genetic haplotypes Human genes 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 239000012577 media supplement Substances 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229950000844 mizoribine Drugs 0.000 description 1
- 238000002715 modification method Methods 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 229940014456 mycophenolate Drugs 0.000 description 1
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 229920001308 poly(aminoacid) Polymers 0.000 description 1
- 229940057838 polyethylene glycol 4000 Drugs 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 239000012090 serum-supplement Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/808—Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/808—Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
- Y10S530/809—Fused cells, e.g. hybridoma
Definitions
- the present invention relates to an anti-histone HI monoclonal antibody and a polypeptide that specifically recognizes the anti-histone HI antibody and the anti-histone HI antibody. More specifically, the present invention relates to a monoclonal antibody and a hyperidoma and a polypeptide that produce the antibody, which are useful for suppressing, predicting or diagnosing rejection in organ transplantation.
- immunosuppressive agents In organ transplantation medicine, various immunosuppressive agents have been used in the past to suppress rejection after organ transplantation. Examples of such immunosuppressive agents include takuguchi limus (FK506) and cyclosporin A (Jpn J Pharmacoljl, 89-100, 1996).
- takuguchi limus FK506
- cyclosporin A Jpn J Pharmacoljl, 89-100, 1996.
- problems such as the promotion of cancer cell proliferation, strong side effects such as suppression of bone marrow function, infection, and the necessity of permanent administration (Transplantation, 58, 170—178, 1994).
- recipient PVG rat serum transplanted with DA rat liver (post-OLT serum) is administered once to a transplantation model system in which a rejection reaction occurs. It has been reported that the reaction is suppressed (J. Surg. Res., 80, 58-61, 1998).
- anti-histone HI polyclonal antibodies are administered postoperatively to DA (RTla) and LWIS lath HRT11) heart transplantation systems (in vivo), where rejection always occurs, and thus rejection is suppressed. It is disclosed that the ent survives (Transplantation, 77, 1595-1603, 2 004).
- MLR mixed lymphocyte reaction
- the present inventors have now found an anti-histone HI monoclonal antibody having a significant immunosuppressive activity and usable for suppressing, predicting or diagnosing transplant rejection, and a hyperidoma producing this antibody. Furthermore, the present inventors have found a specific amino acid sequence that is specifically recognized by the anti-histone HI monoclonal antibody. The present invention is based on strong knowledge.
- the present invention aims to provide an anti-histone HI monoclonal antibody that has remarkable immunosuppressive activity and can be used for suppression, prediction or diagnosis of transplant rejection, and a hyperprideoma producing this antibody.
- Another object of the present invention is to provide a polypeptide comprising a specific amino acid sequence that is specifically recognized by the anti-histone HI monoclonal antibody.
- the monoclonal antibody according to the present invention is characterized by recognizing a histone HI-like antigen present on the cell membrane of histone HI or spleen cells.
- the hyperidoma according to the present invention produces the monoclonal antibody.
- polypeptide according to the present invention comprises a specific amino acid sequence recognized by the monoclonal antibody according to the present invention.
- the monoclonal antibody according to the present invention has remarkable immunosuppressive activity and safety, and can be advantageously used as an excellent immunosuppressive agent. Furthermore, the monoclonal antibody according to the present invention has excellent specificity for a mammalian autoantigen protein, which is an index of transplant rejection, and therefore predicts or diagnoses transplant rejection in mammals in organ transplantation. Therefore, it can be advantageously used.
- the polypeptide according to the present invention when used as an antigen, can induce the production of anti-histone HI antibodies in a living body, and therefore can be advantageously used as an immunosuppressant. Furthermore, the polypeptide according to the present invention can be used to measure the amount of anti-histone HI antibody produced in mammals, and thus is advantageous for predicting or diagnosing transplant rejection in organ transplantation. Can be used.
- FIG. 1 shows the results of MLR inhibitory activity evaluation using a culture supernatant containing an anti-histone HI monoclonal antibody.
- Hypridoma 1F5, Hybridoma 3F2, Hybridoma 15F11, Neubridoma 17C2 or Hypridoma 16G9 has an original deposit date of August 19, 2004.
- Country Tsukuba Sakai Higashi Ibaraki Pref. 1— 1— 1 Central No. 6 receipt number ABP—10409, receipt number FERM ABP—10410, receipt number FERM ABP—10411, receipt number FER Deposited under M ABP—10412 or receipt number FERM ABP—10413.
- the monoclonal antibody according to the present invention is characterized by recognizing histone Hl or histone HI-like antigen present in splenocytes.
- the monoclonal antibody according to the present invention preferably recognizes an epitope in the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8.
- the anti-histone HI monoclonal antibody is a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. It recognizes peptides.
- the polypeptide preferably has a power of about 12 to 150 amino acid residues.
- the anti-histone HI monoclonal antibody is a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. It recognizes peptides.
- the anti-histone HI monoclonal antibody is at least one selected from the group force consisting of hyperpridoma 1F5, hybridoma 3F2, hybridoma 15F11, nob, hybridoma 17C2 and hyperpridoma 16G9. Produced by two hyperidomas.
- Histone Hl means a basic protein that exists in a eukaryotic cell and binds to a nucleosome linker DNA to form a nucleosome.
- histone HI examples are those derived from humans and comprising the amino acid sequence shown in SEQ ID NO: 1, and those derived from Ushi and comprising the amino acid sequence shown in SEQ ID NO: 2. Or those derived from mice and comprising the amino acid sequence shown in SEQ ID NO: 3.
- the "histone HI-like antigen" is also recognized on the cell membrane of spleen cells by monoclonal antibodies produced by Hypridoma 1F5, Hypridoma 3F2, Hybridoma 15F11, Hybridoma 17C2, and Hypridoma 16G9. Antigen.
- This histone HI-like antigen preferably constitutes a part of a protein showing a molecular weight of 31 kD on SDS-PAGE. Examples of such histone HI-like antigens include proteins having at least a part of the amino acid sequence of histone HI derived from mammals.
- the monoclonal antibody according to the present invention can be a chimeric antibody, a humanized antibody, or a fully human antibody, if desired.
- a chimeric antibody (Morrison, S ⁇ ., Oi, VT, “immu noglobulin genes” Academic Press (London), 260-274 (1989) ), A human-type antibody that embeds a complementary determining region (CDR), which is a sequence on the Fv domain directly involved in antigen binding of a mouse monoclonal antibody, in a human antibody frame by CDR grafting technology (Roguska, ML et.AL, Humanization of murine monoclonal an tibodies through variable domain resurfacing, Proc. Natl. Acad. Bci. USA, 91, 969-9 73 (1994)), and as a fully human antibody This is based on Tomizuka, K.
- CDR complementary determining region
- hybridoma 1F5 a hybridoma 3F2, a hybridoma 15F11, a hybridoma 17C2, or a hybridoma 16G9.
- the monoclonal antibody and hybridoma according to the present invention can be produced, for example, as follows. That is, first, the hyperidoma according to the present invention uses a histone HI or histone HI-like antigen or a polypeptide containing these epitopes as a sensitizing antigen, and the characteristics of a mammal immunized with the sensitizing antigen. Cells (immune cells) can be obtained by fusing with myeloma cells of mammals, cloning the resulting hybridomas, and selecting from the hybridomas. The monoclonal antibody according to the present invention can be obtained by culturing the hyperidoma according to the present invention and recovering the antibody produced by the culture.
- Histone HI or histone HI-like antigen used as a sensitizing antigen or a polypeptide containing these epitopes includes, for example, human leukemia bone marrow cells, human cervical cancer HeLa cells, Examples include thymus, thymus liver, and avian red blood cells.
- This sensitizing antigen is prepared in the form of a suspension containing, for example, PBS or physiological saline, and if desired, an adjuvant such as FCA (Freund's complete adjuvant) or KLH (keyhole limpet hemocyanin) and the like. Can be used for processing.
- FCA Red's complete adjuvant
- KLH keyhole limpet hemocyanin
- a general administration method in the art can be used. Specifically, intraperitoneal injection, intrasplenic injection, intramuscular injection, subcutaneous injection, intradermal injection, Examples include oral administration, transmucosal administration, and transdermal administration, and intraperitoneal injection and intrasplenic injection are preferred.
- the interval between doses of sensitizing antigen can be determined appropriately depending on the dose of sensitizing antigen and the type of mammal, for example, several times a month.
- the mammal to be immunized is not particularly limited, but is preferably selected in consideration of compatibility with the myeloma cells used for cell fusion, for example, mouse, rat, hamster, etc.
- spleen cells are preferably used as immune cells.
- the myeloma cells used in the present invention include, for example, P3 (P3X63Ag8.653) (J. Immunol., 123, 1548, 1978), p3-Ul (Current Topics in Microbiology and Immunology, 81, 1—7,1978), NS—1 (Eur. J. Immunol, 6, 511—519, 1976), MPC—11 (Cell, 8, 405-415, 1976), Sp2 / 0—Agl4 (Nature, 276 , 269-270, 1978), FO (J. Immun ol. Meth., 35, 1—21, 1980), S194 (J. Exp. Med., 148, 313—323, 1978), and R2 10 (Nature , 277, 131-133, 1979), etc., preferably P3 or p3-Ul, more preferably P3.
- P3 or p3-Ul more preferably P3.
- Cell fusion between immune cells and myeloma cells can be performed, for example, according to the method of Milstein et al. (Methods Enzymo L, 73, 3-46, 1981). Specifically, cell fusion can be performed, for example, by mixing immune cells and myeloma cells in a medium in the presence of a fusion promoter. In cell fusion, the operation of adding an appropriate medium and centrifuging can be repeated to produce nobridoma.
- Examples of the medium used for cell fusion include RPMI-1640 medium and MEM medium. Examples thereof include media usually used for cell fusion.
- serum supplements such as fetal bovine serum (FBS) can be used in combination as appropriate.
- the cell fusion temperature is preferably 25 to 37 ° C, more preferably 30 to 37 ° C.
- the mixing ratio of myeloma cells and immune cells is preferably about 1: 1 to 1:10.
- Examples of the fusion promoter include polyethylene glycol (PEG), Sendai virus (HVJ), and the like, and preferably PEG.
- PEG polyethylene glycol
- HVJ Sendai virus
- the molecular weight of PEG can be appropriately selected.
- the average molecular weight can be about 1,000 to 6,000.
- the concentration of PEG in the medium is preferably about 30 to 60% (WZV).
- an auxiliary such as dimethyl sulfoxide can be appropriately added to the medium.
- the selection of the hyperidoma according to the present invention is carried out by culturing the hyperidoma obtained by cell fusion in a normal selective medium such as HAT medium and using a normal limiting dilution method, for example, an antibody against histone HI. It can be carried out by screening as an index such as value.
- the culture period in the HAT medium is a time sufficient for the cells (unfused cells) other than the target hyperidoma to die, and can usually be several days to several weeks.
- the thus obtained hyperpridoma according to the present invention can be subcultured in a normal medium and can be stored for a long time in liquid nitrogen.
- the method for recovering the monoclonal antibody according to the present invention includes, for example, a method of culturing a hybridoma according to a conventional method to obtain a culture supernatant strength monoclonal antibody, or a hybridoma compatible with this. Examples thereof include a method for administering an antibody to a mammal and allowing it to proliferate to obtain its ascites monoclonal antibody.
- the former method is preferable for obtaining high-purity antibodies, while the latter method is preferable for producing antibodies in large quantities.
- the monoclonal antibody according to the present invention can be purified with high purity by a method such as salting-out, gel filtration, or affinity chromatography.
- the monoclonal antibody according to the present invention has a remarkable immunosuppressive activity.
- the monoclonal antibody according to the present invention can be used as an immunosuppressant as it is, together with a pharmaceutically acceptable carrier and the like, and can be used as a pharmaceutical composition, particularly as an immunosuppressive composition. Therefore, according to one aspect of the present invention, there is provided an immunosuppressive composition comprising the monoclonal antibody according to the present invention as an active ingredient. According to another aspect of the present invention, there is provided use of a monoclonal antibody according to the present invention in the manufacture of a composition for immunosuppression.
- the immunosuppressive composition according to the present invention is used for the treatment and prevention of rejection by transplantation of organs such as the heart, kidney, liver, bone marrow, and skin, and further for the treatment and prevention of autoimmune diseases and the like.
- organs such as the heart, kidney, liver, bone marrow, and skin
- the immunosuppressive composition according to the present invention can be prepared, for example, by dissolving the monoclonal antibody according to the present invention in physiological saline for injection, distilled water for injection, buffer solution for injection, or the like.
- composition for immunosuppression includes an appropriate solvent, a solubilizer, a preservative, a stabilizer, an emulsifier, a suspension, a soothing agent, an isotonic agent, a buffer, an excipient, Thickeners, colorants, known carriers (various ribosomes, polyamino acid carriers, synthetic polymers, natural polymers, etc.) can be included.
- the immunosuppressive composition according to the present invention can be administered systemically or locally.
- Specific administration methods include infusion, intravenous injection, intramuscular injection, subcutaneous injection, and intradermal. Examples include injection, oral administration, transmucosal administration, and transdermal administration.
- a method of treating a mammal in need of immunosuppression comprising administering to the mammal a therapeutically effective amount of a monoclonal antibody according to the present invention. Law is provided.
- the dose of the monoclonal antibody according to the present invention varies depending on the mammal's condition, age, etc. Usually, 0.05 to 40 mgZ body weight kgZ day, preferably 0.1 to 1. OmgZ body weight kgZ day once or several times. Can be administered separately. The administration can be performed only once, but can be repeated, for example, during 4 weeks.
- the monoclonal antibody according to the present invention specifically reacts with a self-antigen protein that serves as an index of transplant rejection during organ transplantation, transplant rejection in mammals can be used for prediction or diagnosis.
- the self-antigen protein is present in mammals and is preferably produced by Hypridoma 1F5, Hypridoma 3F2, Hybridoma 15F11, Hybridoma 17C2 or Hypridoma 16G9.
- a pharmaceutically acceptable carrier can be added to the composition as desired.
- the transplant rejection preferably occurs after organ transplantation, and more preferably occurs after administration of the immunosuppressant is stopped.
- Another aspect of the present invention provides the use of a monoclonal antibody according to the present invention as a predictive or diagnostic agent for transplant rejection in a mammal.
- a method for predicting or diagnosing transplant rejection in a mammal wherein the level of immunoreactivity between a biological sample derived from a mammal and the monoclonal antibody according to the present invention.
- a method is provided comprising measuring.
- the measured immunoreactivity level is a threshold value set in advance with reference to the immunoreactivity level between the biological sample of a mammal in which transplant rejection has occurred and the monoclonal antibody according to the present invention. Higher than that, predict or diagnose that the risk of transplant rejection is high.
- This threshold value is appropriately determined by those skilled in the art according to the species of mammals and donors, sex, type of transplanted organ, measurement method, and the like. According to the prediction or diagnosis method of the present invention, the physical and economic burden on the patient can be reduced by avoiding unnecessary administration of immunosuppressive agents.
- the biological sample is preferably blood, more preferably serum, and the immunosuppressive agent is not particularly limited as long as it is an immunosuppressive agent used for organ transplantation.
- Alkylating agents such as cyclophosphamide, antimetabolites such as azathioprine, methotrexate, mizoribine, inhibitors of T cell activity such as cyclosporine, tacrolimus, predo-zolone, methylpredo-zolone, mofuethyl mycophenolate, azathioprine Steroids such as basiliximab, lymphocyte surface function inhibitors such as basiliximab, muromonab, or combinations thereof.
- Examples of donors for mammals and transplanted organs include humans, pigs, and baboons, with humans being preferred.
- Examples of organs to be transplanted include liver, heart, kidney, and skin.
- Specific examples of the method for measuring the immunoreactivity level are not particularly limited as long as they use an antigen-antibody reaction. For example, fluorescent antibody method, chemical staining method, enzyme antibody method, ELSA method , Radioinometic, immunoprecipitation, western blot, modified Western plot (such as western, southwestern, northwestern, or western), or protein chip method .
- the immunoreactivity level is determined by fluorescent antibody method, chemical staining method, enzyme antibody method, ELISA, radioimmunoassay, immunoprecipitation method, Western plotting method, Measured by modified Western blot or protein chip method.
- kits for predicting or diagnosing transplant rejection in a mammal comprising at least an anti-histone HI monoclonal antibody.
- the transplant rejection preferably occurs after organ transplantation, more preferably after administration of the immunosuppressive agent is stopped.
- polypeptide according to the present invention comprises, in the amino acid sequence thereof, an epitope that recognizes the anti-histone HI monoclonal antibody according to the present invention as described above. And according to the preferable aspect of this invention, polypeptide consists of an amino acid sequence represented by sequence number 4, sequence number 5, sequence number 6, sequence number 7, or sequence number 8.
- one polypeptide is present in the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8.
- the amino acid sequence can be obtained by substitution, deletion or addition of several amino acids.
- the range of “1 or several” means preferably 1 to 3, more preferably about 1 to 2.
- the polypeptide comprises a partial sequence of the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. It will be.
- the partial sequence is preferably a partial sequence represented by Nos. 6 to 9 in SEQ ID No. 4, a partial sequence represented by Nos. 5 to 9 in SEQ ID No. 5, or a second sequence of SEQ ID No. 6
- the polypeptide comprises the amino acid sequence represented by SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, or a partial sequence thereof. It is what.
- this polypeptide preferably has a power of about 3 to 300 amino acid residues, more preferably about 12 to 150 amino acid residues.
- the polypeptide of the present invention has an amino acid sequence determined on the basis of analysis by a phage display peptide library kit using the monoclonal antibody of the present invention.
- the polypeptide according to the present invention can be synthesized by a known peptide synthesizer or the like based on its amino acid sequence.
- the polypeptide according to the present invention can be used for inducing production of an anti-histone HI antibody in a living body as it is or as a derivative by a known method.
- Sarasuko, histone HI or histone HI-like antigen can also be administered to a living body as a sensitizing antigen to produce an anti-histone HI antibody to exert an immunosuppressive action.
- an immunosuppressive composition comprising as an active ingredient a histone Hl, a histone HI-like antigen or a polypeptide according to the present invention.
- use of a histone Hl, a histone HI-like antigen or a polypeptide according to the present invention in the manufacture of a composition for immunosuppression.
- the immunosuppressive composition can be produced into a dosage form according to the administration method together with a pharmaceutically acceptable carrier.
- a pharmaceutically acceptable carrier for example, when the dosage form is a liquid, an appropriate solution is prepared.
- Agents, solubilizers, preservatives, stabilizers, emulsifiers, suspending agents, soothing agents, isotonic and buffering agents, excipients, thickeners, colorants, known carriers (various ribosomes, poly Amino acids, carriers, synthetic polymers, natural polymers), adjuvants, and the like can be appropriately contained.
- the immunosuppressive composition may be administered by a method that can be used in the art, for example, intraarterial injection, infusion, intravenous injection, intramuscular injection, subcutaneous injection, intradermal. Examples include injection, oral administration, transmucosal administration, and transdermal administration.
- a method for treating a mammal in need of immunosuppression wherein the mammal is treated with a therapeutically effective amount of histone HI, histone HI-like antigen or a polynucleotide according to the present invention.
- a method is provided comprising administering a peptide.
- the therapeutically effective amount varies depending on the severity of the disease state, sex, age, body weight, habits, etc. of the mammal, but for example, the amount of the active ingredient is from 0.005 ⁇ g to 2 gZ body weight kgZ. can do. In addition, by confirming antibody production in mammals, those skilled in the art can appropriately prepare such a dosage plan.
- histone Hl, histone HI-like antigen or the polypeptide according to the present invention specifically reacts with anti-histone HI antibody produced in mammals, it can be used for measurement of the amount of anti-histone HI antibodies in mammals. can do. Therefore, according to another aspect of the present invention, a histone Hl, histone HI-like antigen or a polypeptide according to the present invention is effective for measuring the amount of anti-histone HI antibody in a biological sample derived from a mammal. A composition is provided as an ingredient. Since the anti-histone HI antibody has a function of suppressing transplant rejection as described above, the amount of anti-histone HI antibody in mammals is an index for predicting or diagnosing transplant rejection.
- the yarn and adult can be used for prediction or diagnosis of transplant rejection in mammals.
- a histone Hl, histone HI-like antigen according to the present invention or a polypeptide according to the present invention as a predictive or diagnostic agent for transplant rejection in a mammal.
- the transplant rejection occurs after organ transplantation, more preferably after administration of the immunosuppressant is stopped.
- a method for predicting or diagnosing transplant rejection in a mammal comprising an anti-histone HI antibody in a biological sample in a mammal and histone H1, histone H1-like antigen, or There is provided a method comprising measuring the level of immunoreactivity with a polypeptide according to the invention.
- the measured immunoreactivity level force between the anti-histone HI antibody in the biological sample of the mammal in which transplant rejection occurred, and the histone HI, histone HI-like antigen, or the polypeptide according to the present invention Predict or diagnose that the risk of transplant rejection is low if it is higher than a preset threshold with reference to the immunoreactivity level.
- This threshold value is appropriately determined by those skilled in the art according to the species of mammals and donors, sex, type of transplanted organ, measurement method, and the like. According to the prediction or diagnosis method of the present invention, by avoiding unnecessary administration of immunosuppressive agents, the patient's Physical and economic burden can be reduced.
- the method for measuring the immunoreactivity level are not particularly limited as long as they use an antigen-antibody reaction.
- fluorescent antibody method for example, fluorescent antibody method, chemical staining method, enzyme antibody method, ELSA method , Radioinometics, immunoprecipitation, western blotting, modified Western plots (such as western, southwestern, northwestern, or western), protein chip methods, etc.
- the immunoreactivity level is a fluorescent antibody method, chemical staining method, enzyme antibody method, ELISA, radioimmunoassay, immunoprecipitation method, Western blotting method, Western blot modification method or protein chip method,
- the protein chip method is more preferable.
- the polypeptide of the present invention by carrying the polypeptide of the present invention on a protein chip, transplant rejection in a mammal can be predicted or diagnosed quickly and accurately.
- kits for measuring the amount of anti-histone HI antibody in a biological sample derived from a mammal comprising a histone H1, histone HI-like antigen or
- a kit comprising at least a polypeptide according to the invention.
- the kit is for predicting or diagnosing transplant rejection in a mammal.
- the transplant rejection preferably occurs after organ transplantation, more preferably after administration of the immunosuppressant is stopped.
- the biological samples, immunosuppressive agents, transplanted organs, mammals, and donors of transplanted organs described above can be used for the above-mentioned treatment methods and transplantation rejection prediction using the monoclonal antibody according to the present invention. It is the same as the diagnostic method.
- Untreated PVG rat-derived spleen lymphocytes (responsive cells) and DA rat-derived spleen lymphocytes (stimulatory cells) treated with mitomycin C (Kyowa Hakko Kogyo Co., Ltd.) were used.
- the responding cells were adjusted to 5 ⁇ 10 5 cells ZmL with 10% FCS—RPMI medium, and the stimulating cells were adjusted to 8 ⁇ 10 6 cells ZmL with 10% FCS—RPMI medium.
- MLR Mixed lymphocyte reaction
- the responder cells were adjusted to 5 ⁇ 10 5 cells ZmL with 10% FCS—AIM—V medium, and the stimulator cells were adjusted to 8 ⁇ 10 6 cells / mL with 10% F CS—RPMI medium.
- each of the responsive cell suspension and stimulated cell suspension is seeded in a 96-well round bottom plate (Nunc Brand Products), and anti-histone HI polyclonal IgG (Santa Cruz Biote chonology) Made: 0.1, 0.2, 0.4, 0.8, or 1.6 / z gZ uel) or Usagi IgG (.normal rabbit IgG: banta Cruz Biotechonology: 0.1, 0.2, 0.4, 0.8, or 1.6 ⁇ g / well), and cultured for 2.5 days under conditions of 37 ° C, 5% CO / 95% air
- ConA was added to a concentration of 10 gZmL, and it was confirmed that the stimulating cells had stopped proliferating and that the reaction cells caused cell growth by antigen stimulation. Then, each well was treated in the same manner as in Reference Example 1, and brodoxyuridine (BrdU) incorporated into intracellular DNA using BrdU Labeling & Detection Kit III (Roche Diagnostics). The degree of cell proliferation was measured using the amount as an index.
- MLR MLR was suppressed in a concentration-dependent manner by anti-histone polyclonal antibody. On the other hand, MLR was not suppressed by normal rabbit IgG.
- mice 0.2 mL of a solution in which antigen was dissolved in PBS (antigen concentration: 600 to: LOOOmg / mL) was finally administered intraperitoneally to mice.
- blood was collected from the fundus vein and the antibody titer was measured by ELISA.
- whole blood was collected, and the obtained blood was centrifuged (2000 rpm, 20 minutes) to obtain an antiserum, which was used as a control antiserum in the following experiments.
- the spleen was removed from the rat, and the obtained spleen cells were used for the following cell fusion.
- IMDM IMDM
- FCS manufactured by JRH BIOSCIENCES
- UPPLEMENT (X 50), manufactured by SIGMA) is dissolved in 10 mL of serum-free IMDM medium and diluted 50-fold with IMDM medium containing 10% FCS. ) With lOOmL, 37 ° C / 5% CO
- HT medium HT powder (manufactured by HT MEDIA SUPPLEMENT, SIGMA) in 10 mL of serum-free IMDM medium and dilute 50-fold with IMDM medium containing 10% FCS. ), And cultured for 3 days in a 37 ° CZ5% CO incubator. Every 2-3 days thereafter
- the medium (HT medium) was changed. After confirming cell growth with a microscope, the culture supernatant (about 100 mL) was recovered. The culture supernatant was used to screen for hyperidoma by measuring the titer against histone HI shown below.
- histone H 5mg, calf thymus histone Hl, Roche Diagnostics
- Bicarbonate buffer 100 mM NaHCO-NaOH, pH 9.2 to 9.5
- dilution buffer 10 mM Tris-HCl (pH 8.0), 0.9% (W / V) NaCl, 0.05% (W / V) Tween20
- IgG Biotion-labeled anti-mouse IgG, SIGMA
- the plate was washed 6 times, and a fluorescent substrate buffer (Attophos substrate buffer, manufactured by Roche Diagnostics) was added at 50 mL / well, and the plate was shaded to develop color.
- the fluorescence intensity was measured with CytoFluorll (manufactured by Perceptive).
- Test Example 1 Evaluation test of MLR inhibitory activity using pile supernatant containing HI histone HI monoclonal antibody
- Each hybridoma was cultured using IMDM medium containing 15% FCS 10% HSF (1 ⁇ 10 6 cells / mL). 15 mL of this culture supernatant was centrifuged (2000 g, 2.5 hours) using CENTRIPREP YM-10 (MILLIPORE) to obtain a culture supernatant concentrate containing anti-histone HI monoclonal antibody.
- P VGZPVG represents a mixed culture of PVG lymphocytes that have lost their proliferative ability upon stimulation with mitomycin C and PVG lymphocytes that are reactive cells
- DAZPVG is also a DA lymphocyte that has lost proliferative ability. Represents a mixed culture of PVG lymphocytes as reactive cells.
- Test Example 2 Identification of recognition site of anti-histone H 1 monoclonal antibody 1
- Spleen cells were removed from PVG rats and disrupted according to the method of Weissman (Weissman et al., Science, 239, 10 18-1021, 1988). Splenocytes were collected after centrifuging (1,500 rpm, 5 minutes) with PBS lmL in the spleen cells. The cells were suspended using a syringe and the same amount of 150 mM NaCl was collected. The obtained cell suspension was centrifuged (300 ⁇ g, 10 minutes) to obtain a precipitate and a supernatant. This precipitate was used as an insoluble fraction (containing a nuclear fraction), and the supernatant was used as a soluble fraction containing a cell membrane.
- the insoluble fraction is 5 times the volume (v / v) of the electrophoresis sample.
- Pull up the pull buffer (0.25 M Tris-HCl (pH 6.8) 25 mL, SDS 2. Og, ultrapure water 9 mL, gly cerol 10 mL, BPB 5 mg) and boil for 5 minutes. Kiyoshi was used as a measurement sample.
- EDTA was added to the cell membrane-containing soluble fraction to a final concentration of 5 mM. Of this cell membrane-containing soluble fraction, 100-200 1 ⁇ was recovered. The remaining soluble fraction containing cell membrane was treated by ultracentrifugation (4 ° C, 200000 Xg, 45 minutes).
- the obtained precipitate was used as a cell membrane fraction, and the obtained supernatant was used as a cell membrane-removed soluble fraction.
- 5 times (v / v) sample buffer for electrophoresis was added and boiled for 5 minutes, followed by centrifugation.
- the obtained supernatant was used as a measurement sample.
- HCL solution 1 mM HCL solution is fed to a HiTrap NHS column (HiTrap NHS-activated HP ⁇ Amersham Biosciences AB) at l ⁇ 2mL / min, and then histone HI solution (Histone HI (Roche Diagnostics) 1 0.5 mg, coupling buffer (0.2 M NaHC03, 0.5 M NaCl pH 8.3) ImL was fed at ImL / min, and immediately after feeding, the column was sealed and coupling was performed (15- Half an hour). After coupling, buffer A (0.5 M monoethanolamine, 0.5 M NaCl pH 8.3), nofer B (0.1 M sodium acetate, 0.5 M NaCl pH 4.0) and neutral buffer (1.0 M Tris-HCl, pH 9.0) The inside of the column was washed.
- buffer A 0.5 M monoethanolamine, 0.5 M NaCl pH 8.3
- nofer B 0.1 M sodium acetate, 0.5 M NaCl pH 4.0
- neutral buffer 1.0 M Tris-HCl, pH 9.0
- the anti-histone HI monoclonal antibody-containing culture supernatant obtained in Test Example 1 was fed into the column at 1 mL / min and circulated (4 ° C, overnight) 0
- Each measurement sample was processed by SDS-PAGE according to the electrophoresis method (Nature, 227, 680-685, 1970) in the Laemmli discontinuous buffer system.
- Histone Hl 5 mg, manufactured by Roche Diagno Status
- the resulting gel was used for the following Coomassie staining or Western blotting.
- the gel after SDS-PAGE was transferred to a PV DF membrane using a semi-dry transfer device (AE-6675, manufactured by ATTO).
- AE-6675 manufactured by ATTO
- the PVDF membrane after transfer was soaked in a blocking solution (5% skim milk 1% BSA in PBST) and shaken (at 4 ° C. overnight or at room temperature for 1 hour).
- the PVDF membrane was mixed with a solution containing purified anti-histone HI monoclonal antibody as a primary antibody (500-fold diluted with blocking solution), shaken at room temperature for 1 hour, and then PBST Washed for 15 min X l times and 5 min X 3 times.
- a secondary antibody HRP-anti-mouse IgG (manufactured by SIGMA)
- HRP-anti-mouse IgG manufactured by SIGMA
- a blocking solution was added to the PVDF membrane and shaken at room temperature for 1 hour. After shaking, the plate was washed with PBST for 15 minutes X I times and 5 minutes X 3 times.
- the ECL Western blotting detection system was used to detect the binding of specific antibodies and applied to the X-ray film RX-U (FUJI PHOTO FILM, Tokyo, Japan).
- PVG rat spleen cells were suspended in PBS solution containing 4% formalin and fixed at room temperature for 20 minutes. Further, the spleen cells were washed 3 times with a staining buffer (Staining buffer: 1% ( ⁇ / ⁇ ) FCS, 0.1% (w / v) Sodium azide-containing PBS solution, 4 ° C), and then stained. A mixture containing 2 ⁇ 10 6 cells in 100 mL of buffer was prepared. A primary antibody (2 mL of piotin-labeled anti-histone HI monoclonal antibody or 5 mL of normal mouse IgG labeled with piotin) was added to this mixture and reacted at 37 ° C for 1 hour.
- Staining buffer 1% ( ⁇ / ⁇ ) FCS, 0.1% (w / v) Sodium azide-containing PBS solution, 4 ° C
- the spleen cells were washed 3 times with a staining buffer (4 ° C), and further, the staining buffer lOOmL was added to the spleen cells, and FITC-labeled streptavidin (BD PharMingen) lmL was collected. The reaction was allowed to proceed for 30 hours at room temperature. After the reaction, the spleen cells were washed 3 times with a staining buffer (4 ° C), and 500 mL of PBS was added to the spleen cells to prepare a suspension. To this suspension, Propidum Iodide (SIGMA) was added to a final concentration of 5 mgZmL and allowed to react at room temperature for 20 minutes. The obtained cells were sealed with a PBS solution containing 50% glycerin and then observed with a fluorescence microscope.
- SIGMA Propidum Iodide
- peripheral (cell membrane) part of the spleen cells was specifically fluorescently stained.
- Test, Test Example 4 Pile histone HI monoclonal pile recognition site identification 3
- TBST 50 mM Tris, 150 mM NaCl, 0.1% Tween 20. 4 x 10 1 in the original library on the first Panjung. Individual phages were used for the staring jung. Unbound phages were removed by repeated washing with TBTS. The bound phage was eluted with 0.2M Glycine-HC1 (pH 2.2), lmg / ml BSA. The eluted phage was grown in 20 mL E Coli ER2738 culture. Obtained The resulting phages were precipitated using polyethylene glycol and used for the second panning. In addition, a third panning was performed according to the same operating procedure.
- Plaques obtained in the third round of panning were diluted 1: 100 and grown using ER2738 culture. The tubes containing these were incubated at 37 ° C with shaking for 4.5-5 hours. Single-stranded phage DNA was precipitated and purified with Iodide buffer (10 mM Tris-HC1, ImM EDTA, 4M Nal) and ethanol. Phage DNA was dissolved in 20 ⁇ L ⁇ buffer (10 mM Tris—HC1 pH 8.0, ImM EDTA) for DNA sequence analysis.
- 3F2 VTNNQTSPRWEI (SEQ ID NO: 5)
- Each peptide having an amino acid sequence determined from the phage DNA was synthesized by a conventional method. Competition ELISA was performed using the obtained peptide, each purified monoclonal antibody, and histone HI antigen (Roche, catalog # 1004875). At this time, EZ-Link Sulfo-NHS-Biotinylation kit (Pierce) was used for the biotinylation of histone HI antigen. ABTS solution (Sigma, A3219) was used as a reagent. Color development was detected at 405 nm using an ELISA measuring device (ThermoLabsyst em, Multiskan Ascent). The measured value was the average of the absorption values measured three times.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Transplantation (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2005800380989A CN101056979B (zh) | 2004-09-03 | 2005-09-05 | 抗组蛋白h1单克隆抗体及用于产生其的杂交瘤 |
US11/661,763 US7964554B2 (en) | 2004-09-03 | 2005-09-05 | Polypeptide that binds anti-histone H1 antibody |
EP05777004A EP1820855A4 (en) | 2004-09-03 | 2005-09-05 | MONOCLONAL ANTI-HISTON-H1 ANTIBODY AND ITS MANUFACTURE OF HYBRIDOMA |
JP2006532019A JP4855261B2 (ja) | 2004-09-03 | 2005-09-05 | 抗ヒストンh1モノクローナル抗体およびこれを産生するハイブリドーマ |
US12/762,649 US8187602B2 (en) | 2004-09-03 | 2010-04-19 | Anti-histone H1 monoclonal antibody and hybridoma for the production thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004-257528 | 2004-09-03 | ||
JP2004257528 | 2004-09-03 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/661,763 A-371-Of-International US7964554B2 (en) | 2004-09-03 | 2005-09-05 | Polypeptide that binds anti-histone H1 antibody |
US12/762,649 Division US8187602B2 (en) | 2004-09-03 | 2010-04-19 | Anti-histone H1 monoclonal antibody and hybridoma for the production thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2006025580A1 true WO2006025580A1 (ja) | 2006-03-09 |
Family
ID=36000214
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/016268 WO2006025580A1 (ja) | 2004-09-03 | 2005-09-05 | 抗ヒストンh1モノクローナル抗体およびこれを産生するハイブリドーマ |
Country Status (6)
Country | Link |
---|---|
US (2) | US7964554B2 (ja) |
EP (1) | EP1820855A4 (ja) |
JP (1) | JP4855261B2 (ja) |
KR (1) | KR20070073754A (ja) |
CN (1) | CN101056979B (ja) |
WO (1) | WO2006025580A1 (ja) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009001673A1 (ja) * | 2007-06-26 | 2008-12-31 | Josai University Corporation | Atp含有免疫アジュバント |
WO2009004900A1 (ja) * | 2007-06-29 | 2009-01-08 | Josai University Corporation | ユビキノン含有免疫アジュバント |
WO2009044555A1 (ja) * | 2007-10-04 | 2009-04-09 | Josai University Corporation | 製剤、当該製剤を用いたワクチンの投与方法及びイオントフォレーシス装置 |
WO2012023614A1 (ja) | 2010-08-20 | 2012-02-23 | 学校法人 城西大学 | 免疫抑制活性を有するモノクローナル抗体またはその抗原結合断片 |
WO2013125687A1 (ja) | 2012-02-22 | 2013-08-29 | 学校法人 城西大学 | 炎症疾患治療剤 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150077184A (ko) | 2013-12-27 | 2015-07-07 | 삼성전자주식회사 | 의료 영상의 병변 유사도 판단 장치 및 방법 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004149507A (ja) * | 2002-09-05 | 2004-05-27 | Hisamitsu Pharmaceut Co Inc | 免疫抑制剤 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6033864A (en) | 1996-04-12 | 2000-03-07 | The Regents Of The University Of California | Diagnosis, prevention and treatment of ulcerative colitis, and clinical subtypes thereof, using microbial UC pANCA antigens |
KR100261114B1 (ko) | 1998-01-24 | 2000-07-01 | 박종헌 | 히스톤을 함유하는 류마티스 관절염 치료제 조성물 |
WO2002051865A2 (fr) * | 2000-12-22 | 2002-07-04 | Aventis Pasteur | Antigenes proteiques inducteurs d'anticorps neutralisant le virus vih |
CN1459505A (zh) * | 2002-05-21 | 2003-12-03 | 赵平 | 中国汉族人基质金属蛋白酶-9基因重构、表达、纯化及其医学应用 |
US20040052780A1 (en) * | 2002-09-05 | 2004-03-18 | Hisamitsu Pharmaceutical Co., Inc. | Immunosuppresants |
-
2005
- 2005-09-05 US US11/661,763 patent/US7964554B2/en not_active Expired - Fee Related
- 2005-09-05 WO PCT/JP2005/016268 patent/WO2006025580A1/ja active Application Filing
- 2005-09-05 CN CN2005800380989A patent/CN101056979B/zh not_active Expired - Fee Related
- 2005-09-05 EP EP05777004A patent/EP1820855A4/en not_active Withdrawn
- 2005-09-05 KR KR1020077006607A patent/KR20070073754A/ko not_active Application Discontinuation
- 2005-09-05 JP JP2006532019A patent/JP4855261B2/ja active Active
-
2010
- 2010-04-19 US US12/762,649 patent/US8187602B2/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004149507A (ja) * | 2002-09-05 | 2004-05-27 | Hisamitsu Pharmaceut Co Inc | 免疫抑制剤 |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009001673A1 (ja) * | 2007-06-26 | 2008-12-31 | Josai University Corporation | Atp含有免疫アジュバント |
JPWO2009001673A1 (ja) * | 2007-06-26 | 2010-08-26 | 学校法人 城西大学 | Atp含有免疫アジュバント |
WO2009004900A1 (ja) * | 2007-06-29 | 2009-01-08 | Josai University Corporation | ユビキノン含有免疫アジュバント |
JPWO2009004900A1 (ja) * | 2007-06-29 | 2010-08-26 | 学校法人 城西大学 | ユビキノン含有免疫アジュバント |
WO2009044555A1 (ja) * | 2007-10-04 | 2009-04-09 | Josai University Corporation | 製剤、当該製剤を用いたワクチンの投与方法及びイオントフォレーシス装置 |
JPWO2009044555A1 (ja) * | 2007-10-04 | 2011-02-03 | 学校法人 城西大学 | 製剤、当該製剤を用いたワクチンの投与方法及びイオントフォレーシス装置 |
WO2012023614A1 (ja) | 2010-08-20 | 2012-02-23 | 学校法人 城西大学 | 免疫抑制活性を有するモノクローナル抗体またはその抗原結合断片 |
KR20130137612A (ko) * | 2010-08-20 | 2013-12-17 | 각코우호우진 조사이 다이가쿠 | 면역 억제 활성을 갖는 모노클로날 항체 또는 그것의 항원 결합 단편 |
RU2559550C2 (ru) * | 2010-08-20 | 2015-08-10 | Дзесаи Юниверсити Корпорейшн | Моноклональное антитело, имеющее иммуносупрессивную активность, или его антигенсвязывающий фрагмент |
JP5960596B2 (ja) * | 2010-08-20 | 2016-08-02 | 学校法人 城西大学 | 免疫抑制活性を有するモノクローナル抗体またはその抗原結合断片 |
US9701739B2 (en) | 2010-08-20 | 2017-07-11 | Josai University Corporation | Monoclonal antibody having immunosuppressive activity or antigen binding fragment thereof |
KR101863978B1 (ko) | 2010-08-20 | 2018-06-01 | 각코우호우진 조사이 다이가쿠 | 면역 억제 활성을 갖는 모노클로날 항체 또는 그것의 항원 결합 단편 |
WO2013125687A1 (ja) | 2012-02-22 | 2013-08-29 | 学校法人 城西大学 | 炎症疾患治療剤 |
Also Published As
Publication number | Publication date |
---|---|
US20080287352A1 (en) | 2008-11-20 |
EP1820855A4 (en) | 2010-05-12 |
JPWO2006025580A1 (ja) | 2008-05-08 |
CN101056979B (zh) | 2013-06-05 |
EP1820855A1 (en) | 2007-08-22 |
US20100196384A1 (en) | 2010-08-05 |
KR20070073754A (ko) | 2007-07-10 |
US7964554B2 (en) | 2011-06-21 |
JP4855261B2 (ja) | 2012-01-18 |
CN101056979A (zh) | 2007-10-17 |
US8187602B2 (en) | 2012-05-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
TW200936160A (en) | Monoclonal antibodies that bind to anexelekto and uses thereof | |
US20040047880A1 (en) | Component for vaccine | |
US8187602B2 (en) | Anti-histone H1 monoclonal antibody and hybridoma for the production thereof | |
US20220098290A1 (en) | Antibodies that bind to cleaved form of mutant calreticulin, and diagnostic, preventive, or therapeutic agent for myeloproliferative neoplasm | |
WO2018105560A1 (ja) | Claudin 5抗体、及びその抗体を含有する医薬 | |
US9309307B2 (en) | Antibody against amyloid precursor protein signal peptide | |
CN103534271B (zh) | 有免疫抑制活性的单克隆抗体或其抗原结合片段 | |
EP3731861A1 (en) | T cell receptors for tumor specific proteasome splice variants and uses thereof | |
CN113710700A (zh) | Rubicon RUN结构域的苏氨酸166和丝氨酸189作为LRRK2激酶抑制靶点 | |
WO1991012332A1 (fr) | Anticorps monoclonaux reconnaissant un peptide associe a un antigene majeur d'histocompatibilite | |
JP4651495B2 (ja) | Isg15タンパク質と特異的に反応するモノクローナル抗体及びそれを産生するハイブリドーマ並びに癌およびウイルス感染の検出方法 | |
JP2015117181A (ja) | 敗血症治療剤 | |
US5681700A (en) | Assay for pathogenicity of anti-DNA antibodies | |
US6433148B1 (en) | Monoclonal anti-idiotypic antibodies (AB2) and their uses | |
CN114605553B (zh) | 含有Treg细胞的药物组合物以及在自身免疫性疾病中的用途 | |
WO2022068895A1 (zh) | 抗SARS-CoV-2刺突蛋白膜外区的单克隆抗体及其应用 | |
JP2004149507A (ja) | 免疫抑制剤 | |
EP3947472A1 (en) | Therapeutic antibodies for treating lung cancer | |
CN114349857A (zh) | 一种Treg细胞制备方法以及在自身免疫性疾病方面的应用 | |
JP2004155745A (ja) | ペプチドおよびその利用 | |
JP2008056647A (ja) | Fcα/μレセプターに対する抗体 | |
JPWO2013125687A1 (ja) | 炎症疾患治療剤 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU LV MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2006532019 Country of ref document: JP |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1020077006607 Country of ref document: KR |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005777004 Country of ref document: EP Ref document number: 11661763 Country of ref document: US Ref document number: 2516/DELNP/2007 Country of ref document: IN |
|
WWE | Wipo information: entry into national phase |
Ref document number: 200580038098.9 Country of ref document: CN |
|
WWP | Wipo information: published in national office |
Ref document number: 2005777004 Country of ref document: EP |