DE4326573A1 - Polypeptides encoded by exon v5 of the CD44 gene as targets for immunotherapy and immunoscintigraphy of tumors - Google Patents
Polypeptides encoded by exon v5 of the CD44 gene as targets for immunotherapy and immunoscintigraphy of tumorsInfo
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- DE4326573A1 DE4326573A1 DE4326573A DE4326573A DE4326573A1 DE 4326573 A1 DE4326573 A1 DE 4326573A1 DE 4326573 A DE4326573 A DE 4326573A DE 4326573 A DE4326573 A DE 4326573A DE 4326573 A1 DE4326573 A1 DE 4326573A1
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Classifications
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- 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
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2884—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD44
-
- 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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
Die Erfindung betrifft die Verwendung von Antikörpern gegen bestimmte tumorassoziierte Antigene zur Behandlung oder zur in-vivo-Diagnostik von Tumoren, Mittel, die zur Be handlung oder in-vivo-Diagnostik von Tumoren geeignet sind, sowie die Verwendung von Polypeptiden, die durch Exon v5 kodierte Sequenzen enthalten, oder v5-spezifischen anti idiotypischen Antikörpern als Tumorvakzine.The invention relates to the use of antibodies against certain tumor-associated Antigens for the treatment or for in vivo diagnosis of tumors, agents which are used for the treatment or in vivo diagnosis of tumors are suitable, as well as the use of Polypeptides containing sequences encoded by exon v5 or v5-specific anti idiotypic antibodies as tumor vaccines.
Seit der Einführung der Hybridomatechnologie im Jahre 1975 (Köhler et Milstein) wurde intensiv nach tumorassoziierten bzw. -spezifischen Antigenen gesucht. Per definitionem wird ein tumorspezifisches Antigen in einer bestimmten malignen Erkrankung exprimiert, während es in anderen Tumortypen sowie in normalen adulten und fötalen Geweben abwe send ist. Für den Einsatz tumorspezifischer Antikörper in der Tumortherapie sowie zum Imaging von Tumoren ist dabei der zweite Aspekt, die Abwesenheit im Normalgewebe, ent scheidend. Bis heute wurde kein tumorspezifischer Marker gefunden, der die Definition voll erfüllt. Eine ganze Reihe von Antikörpern gegen tumorassoziierte Antigene wurde bis jetzt entwickelt und in klinischen Studien zur Behandlung von Tumoren oder zum Imaging von Tumoren eingesetzt (Mulshine et al., 1991; Vitetta et Thorpe, 1991; Larson et al., 1991). Das Prinzip dabei ist, daß der Antikörper als Sonde spezifisch an Tumorzellen bindet und entweder direkt oder über ein an ihn gekoppeltes Agens die Tumorzelle zerstört oder über den Nachweis eines mit dem Antikörper verknüpften Labels zur Darstellung des Tumors ge nutzt werden kann. Die mit dem Antikörper verknüpften Agentien können selbst zyto toxisch sein, beispielsweise radioaktive Isotope, bakterielle oder pflanzliche Toxine oder Zytostatika, immunmodulatorisch wirken wie z. B. Zytokine, oder auch beispielsweise En zyme sein, die ein Vorläufermolekül (Prodrug) in ein zytotoxisches Agens umwandeln kön nen. Bei der visuellen Darstellung von Tumoren in vivo (Imaging) können radioaktiv mar kierte Antikörper verwendet werden; seit einiger Zeit werden auch Kernspinresonanzmetho den entwickelt, bei denen z. B. mit chelatisierten dreiwertigen Kationen verknüpfte Antikör per als Kontrastmittel verwendet werden (Brasch, 1992; Hider et Hall, 1991; Saccavini et al., 1988). Bis heute konnte allerdings kein Antigen/Antikörperpaar gefunden werden, das eine breite Anwendung in der Tumortherapie und in-vivo-Diagnostik erlaubt. Die Suche nach tumorassoziierten Antigenen als geeignete Targets für Antikörper-gestützte Methoden der Tumortherapie und -diagnostik hält daher unvermindert an.Since the introduction of hybridoma technology in 1975 (Köhler et Milstein) intensively searched for tumor-associated or -specific antigens. By definition a tumor-specific antigen is expressed in a specific malignant disease, while it deviates in other types of tumors as well as in normal adult and fetal tissues is send. For the use of tumor-specific antibodies in tumor therapy and for Imaging of tumors is the second aspect, the absence in normal tissue outgoing. To date, no tumor-specific marker has been found that fully meets the definition Fulfills. A whole host of antibodies to tumor-associated antigens have so far been developed developed and in clinical studies for the treatment of tumors or for imaging Tumors used (Mulshine et al., 1991; Vitetta et Thorpe, 1991; Larson et al., 1991). The principle is that the antibody specifically binds to tumor cells and either directly or via an agent coupled to it, destroys the tumor cell or via the detection of a label linked to the antibody to represent the tumor can be used. The agents linked to the antibody can themselves be cytotoxic be toxic, for example radioactive isotopes, bacterial or vegetable toxins or Cytostatics, immunomodulatory such. B. cytokines, or for example En be enzymes that can convert a precursor molecule (prodrug) into a cytotoxic agent nen. When visualizing tumors in vivo (imaging), radioactive mar labeled antibodies are used; for some time also nuclear magnetic resonance methods the developed where z. B. linked antibody with chelated trivalent cations can be used as a contrast medium (Brasch, 1992; Hider et Hall, 1991; Saccavini et al., 1988). To date, however, no antigen / antibody pair has been found that wide use in tumor therapy and in vivo diagnostics allowed. The search for tumor-associated antigens as suitable targets for antibody-based methods tumor therapy and diagnosis therefore continues unabated.
Es wurde kürzlich gezeigt, daß die Expression von Varianten des Oberflächen-Glykopro teins CD44 notwendig und hinreichend ist, um sogenanntes spontanes metastatisches Ver halten sowohl in einer nicht-metastasierenden Pankreas-Adenokarzinom-Zellinie der Ratte als auch in einer nicht-metastasierenden Fibrosarkom-Zellinie der Ratte auszulösen (Günthert et al., 1991). Während die kleinste CD44-Isoform, die Standardform CD44s, in einer Reihe verschiedener Gewebe, darunter Epithelzellen, ubiquitär exprimiert wird, wer den bestimmte Spleißvarianten von CD44 (CD44v) nur auf einer Untergruppe von Epithel zellen exprimiert. Die CD44-Varianten werden durch alternatives Spleißen so erzeugt, daß die Sequenzen von 10 Exons (v1-v10) in CD44s komplett ausgeschnitten werden, jedoch in den größeren Varianten in verschiedenen Kombinationen vorkommen können (Screaton et al., 1992; Tölg et al., 1993; Hofmann et al., 1991). Die Varianten unterscheiden sich dadurch, daß an einer bestimmten Stelle des extrazellulären Teils des Proteins unterschied liche Aminosäuresequenzen inseriert sind. Solche Varianten konnten in verschiedenen menschlichen Tumorzellen und in menschlichem Tumorgewebe nachgewiesen werden. So wurde kürzlich die Expression von CD44-Varianten im Verlauf der kolorektalen Karzinoge nese untersucht (Heider et al., 1993). Die Expression von CD44-Varianten fehlt in norma lem menschlichem Kolonepithel und nur eine schwache Expression ist in den proliferieren den Zellen der Krypten nachweisbar. In späteren Stadien der Tumorprogression, z. B. in Adenokarzinomen, exprimieren alle malignen Entartungen Varianten von CD44. Gewebs expression von variantem CD44 auf hohem Niveau konnte auch in aggressiven Non- Hodgkin-Lymphomen gezeigt werden (Koopman et al., 1993).It has recently been shown that expression of variants of the surface glycopro teins CD44 is necessary and sufficient to avoid so-called spontaneous metastatic ver hold in a non-metastatic pancreatic adenocarcinoma cell line of the rat as well as in a non-metastatic fibrosarcoma cell line of the rat (Günthert et al., 1991). While the smallest CD44 isoform, the standard form CD44s, in a number of different tissues, including epithelial cells, are ubiquitously expressed the specific splice variants of CD44 (CD44v) only on a subset of epithelium cells expressed. The CD44 variants are produced by alternative splicing in such a way that the sequences of 10 exons (v1-v10) are completely cut out in CD44s, however in the larger variants can occur in different combinations (Screaton et al., 1992; Tölg et al., 1993; Hofmann et al., 1991). The variants differ characterized in that at a certain point in the extracellular part of the protein amino acid sequences are inserted. Such variants could be in different human tumor cells and in human tumor tissue are detected. So was recently the expression of CD44 variants in the course of colorectal carcinogen nese examined (Heider et al., 1993). The expression of CD44 variants is missing in norma Human colonic epithelium and only weak expression is proliferating in the detectable in the cells of the crypts. In later stages of tumor progression, e.g. B. in Adenocarcinomas, all malignancies express variants of CD44. Tissue expression of variant CD44 at a high level could also be achieved in aggressive non- Hodgkin lymphomas are shown (Koopman et al., 1993).
Aufgabe der vorliegenden Erfindung war es, neue Mittel zur Behandlung oder zur in-vivo- Diagnostik von Krebserkrankungen bereitzustellen, die auf dem Prinzip der immunologi schen Erkennung tumorassoziierter Antigene beruhen.The object of the present invention was to develop new agents for treatment or for in vivo To provide diagnosis of cancer based on the principle of immunology based detection of tumor-associated antigens.
Diese Aufgabe konnte mit der vorliegenden Erfindung gelöst werden. Die erfindungsgemä ßen Mittel sowie deren Verwendung beruhen auf Antikörpern gegen Epitope, die durch va riante Exons des CD44-Gens kodiert werden, insbesondere von Antikörpern gegen Epitope, die durch das variante Exon v5 des menschlichen CD44-Gens kodiert werden, sowie auf v5- spezifischen antiidiotypischen Antikörpern. Die Antikörper können polyklonal oder mono klonal sein, komplette Immunglobuline, Fab- oder F(ab′)₂-Fragmente von Immunglobulinen oder andere Derivate, bispezifische, chimäre oder humanisierte Antikörper oder rekombinant hergestellte Antikörper, z. B. single-chain-Antikörper (scFv), Fab-Fragmente, andere Frag mente oder komplette Immunglobuline. Die Antikörper können ohne zusätzliches Agens appliziert werden oder mit einer radioaktiven Substanz, einem zytotoxischen Agens, einem immunmodulierenden Agens, einer Substanz, mit deren Hilfe ein zytotoxisches Agens lokal erzeugt werden kann, einem Kernspinresonanz-Kontrastmittel oder einem anderen detek tierbaren Label verknüpft sein. Die Erfindung betrifft ferner die Verwendung von Polypepti den, die Sequenzen enthalten, die durch das variante Exon v5 des CD44-Gens kodiert wer den, und v5-spezifischen antiidiotypischen Antikörpern als Tumorvakzine.This object was achieved with the present invention. The invention ß agents and their use are based on antibodies against epitopes, which by va riante exons of the CD44 gene are encoded, in particular of antibodies against epitopes, which are encoded by the variant exon v5 of the human CD44 gene, as well as on v5- specific anti-idiotypic antibodies. The antibodies can be polyclonal or mono be clonal, complete immunoglobulins, Fab or F (ab ′) ₂ fragments of immunoglobulins or other derivatives, bispecific, chimeric or humanized antibodies or recombinant produced antibodies, e.g. B. single chain antibody (scFv), Fab fragments, other frag elements or complete immunoglobulins. The antibodies can be used without an additional agent be applied or with a radioactive substance, a cytotoxic agent, a immunomodulating agent, a substance with the help of which a cytotoxic agent locally can be generated, a magnetic resonance contrast agent or another detek animal label. The invention further relates to the use of polypepti those that contain sequences that are encoded by the variant exon v5 of the CD44 gene the, and v5-specific anti-idiotypic antibodies as tumor vaccines.
Trägt der Antikörper ein detektierbares Label, kann eine Detektion des Labels zu diagnosti schen Zwecken, z. B. Visualisierung des Tumores in vivo (Imaging), oder beispielsweise zur radiounterstützen Chirurgie (radioguided surgery) erfolgen.If the antibody bears a detectable label, the label can be diagnosed purposes, e.g. B. visualization of the tumor in vivo (imaging), or for example radio-assisted surgery (radioguided surgery).
Die Nuklein- und Aminosäuresequenz des varianten Teils des CD44-Gens ist bekannt (Hofmann et al., 1991; Screaton et al., 1992; Tölg et al., 1993). Die Existenz degenerierter oder alleler Varianten ist für die Ausführung der Erfindung nicht von Bedeutung; solche Varianten sind daher ausdrücklich mit eingeschlossen. Die Sequenz von Exon v5The nucleic and amino acid sequence of the variant part of the CD44 gene is known (Hofmann et al., 1991; Screaton et al., 1992; Tölg et al., 1993). The existence of degenerate or allelic variants is not important for the implementation of the invention; such Variants are therefore expressly included. The sequence of exon v5
ist besonders bevorzugt. Als Mittel, um die Erfindung auszuführen, können Antikörper dienen, insbesondere solche, die gegen Epitope innerhalb der Sequenz von Exon v5 gerich tet sind, oder v5-spezifische antiidiotypische Antikörper. Besonders bevorzugt sind mono klonale Antikörper. Für das erfindungsgemäße Verfahren können jedoch auch polyklonale Antikörper, Fab- oder F(ab′)₂-Fragmente von Immunglobulinen, rekombinant hergestellte Antikörper oder -fragmente, z. B. single-chain-Antikörper (scFv), bispezifische, chimäre oder humanisierte Antikörper oder äquivalente Moleküle dienen, die Exon-v5-kodierte Epitope spezifisch binden. Die Herstellung von Antikörpern gegen bekannte Aminosäurese quenzen kann nach an sich bekannten Methoden erfolgen (Catty, 1989). Beispielsweise kann ein Peptid dieser Sequenz synthetisch hergestellt und als Antigen in einem Immunisie rungsprotokoll eingesetzt werden. Ein anderer Weg ist die Herstellung eines Fusionspro teins, das die gewünschte Aminosäuresequenz enthält, indem eine Nukleinsäure (die syn thetisch oder z. B. durch Polymerase-Kettenreaktion (PCR) aus einer geeigneten Probe hergestellt werden kann), die für diese Sequenz kodiert, in einen Expressionsvektor inte griert und das Fusionsprotein in einem Wirtsorganismus exprimiert wird. Das gegebenen falls gereinigte Fusionsprotein kann dann als Antigen in einem Immunisierungsprotokoll eingesetzt und Insert-spezifische Antikörper oder, im Falle monoklonaler Antikörper, Hy bridome, die insertspezifische Antikörper exprimieren, mit geeigneten Verfahren selektiert werden (Wunderlich et al., 1992). Das komplette CD44-Gen oder ein Fragment davon (z. B. der extrazelluläre Anteil) können auch in einem geeigneten System exprimiert werden, ohne daß die exprimierten Sequenzen mit anderen Peptiden fusioniert sind, dann isoliert und als Antigene in Immunisierungsprotokollen eingesetzt werden. Solche Verfahren sind Stand der Technik. Heider et al. (1993) und Koopman et al. (1993) beschreiben die Herstellung von Antikörpern gegen variante Epitope von CD44.is particularly preferred. Antibodies can be used as a means to practice the invention serve, especially those that are directed against epitopes within the sequence of exon v5 tet, or v5-specific anti-idiotypic antibodies. Mono are particularly preferred clonal antibodies. However, polyclonal can also be used for the method according to the invention Antibodies, Fab or F (ab ′) ₂ fragments of immunoglobulins, recombinantly produced Antibodies or fragments, e.g. B. single-chain antibody (scFv), bispecific, chimeric or humanized antibodies or equivalent molecules that encode exon v5 Bind epitopes specifically. The production of antibodies against known amino acids sequences can be carried out according to methods known per se (Catty, 1989). For example A peptide of this sequence can be synthesized and used as an antigen in an immunization protocol are used. Another way is to make a fusion pro teins containing the desired amino acid sequence by adding a nucleic acid (the syn theoretical or z. B. by polymerase chain reaction (PCR) from a suitable sample can be produced), which codes for this sequence, into an expression vector inte freezes and the fusion protein is expressed in a host organism. The given if purified fusion protein can then be used as an antigen in an immunization protocol used and insert-specific antibodies or, in the case of monoclonal antibodies, Hy bridomes that express insert-specific antibodies are selected using suitable methods (Wunderlich et al., 1992). The entire CD44 gene or a fragment thereof (e.g. the extracellular portion) can also be expressed in a suitable system without that the expressed sequences are fused to other peptides, then isolated and as Antigens can be used in immunization protocols. Such procedures are state of the art Technology. Heider et al. (1993) and Koopman et al. (1993) describe the production of Antibodies against variant epitopes of CD44.
Statt eines intakten Immunglobulinmoleküls können auch Fab- oder F(ab′)₂-Fragmente oder andere Fragmente verwendet werden (Kreitman et al., 1993). Weiter können chimäre Anti körper verwendet werden, z. B. humanisierte Maus-Antikörper (Shin et al., 1989; Güssow et Seemann, 1991), bispezifische Antikörper (Weiner et al., 1993; Goodwin, 1989) oder single-chain-Antikörper/Toxin-Fusionsproteine (Friedman et al., 1993). Die Antikörper können aus Seren oder aus Hybridomaüberständen isoliert oder durch rekombinante Ex pression als single-chain-Antiköper (scFv, Johnson et Bird, 1991), komplette oder fragmen tarische Immunglobuline, gewonnen werden (Coloma et al., 1992; Nesbit et al., 1992; Bar bas et al., 1992). Die Antikörper können allein oder verknüpft mit einem Agens eingesetzt werden. Soll der Antikörper therapeutisch oder für die Immunszintigraphie verwendet werden kann er mit einem geeigneten radioaktiven Isotop z. B. ¹³¹I, ¹²⁵I, ¹¹¹In, ¹⁸⁶Re, ⁹⁰Y, 99mTc oder ²¹¹At verknüpft sein. Diese Verknüpfung kann direkt oder über ein Linker-Mole kül, z. B. einen Chelatbildner, erfolgen. Methoden der Radiomarkierung von Antikörpern sind Stand der Technik (Larson et al., 1991; Thomas et al., 1989; Greiner et al., 1993; Sri vastava, 1988; Rhodes et al., 1986). Für die therapeutische Verwendung kann der Antikör per auch mit einem nicht radioaktiven zytotoxischen Agens verknüpft sein. Dies kann ein Zytostatikum (Schrappe et al., 1992) oder ein zytotoxisches Polypeptid, z. B. ein bakteriel les oder pflanzliches Toxin, sein (Vitetta et al., 1991; Kreitman et al., 1993). Ein solches zytotoxisches Polypeptid kann kovalent, z. B. über Disulfidbrücken, mit dem Antikörper verknüpft sein (Theuer et al., 1993), oder in Form eines Fusionsproteins in einem einketti gen Immunotoxin (Chaudhary et al., 1990; Friedman et al., 1993) mit einem Antikörper verbunden sein. Der Antikörper kann ferner mit einem Zytokin oder einem anderen immun modulatorischen Polypeptid verknüpft sein, z. B. mit Tumornekrosefaktor oder Interleu kin-2. Der Antikörper kann auch mit einem Agens verknüpft sein, das zwar selbst nicht zytotoxisch ist, jedoch eine zytotoxische Substanz erzeugen kann, z. B. ein Enzym, das die Umwandlung eines inaktiven Vorläufermoleküls (Prodrug) in ein Zytostatikum katalysiert (Wang et al., 1992; Senteretal., 1989).Instead of an intact immunoglobulin molecule, Fab or F (ab ′) ₂ fragments or other fragments can also be used (Kreitman et al., 1993). Chimeric anti bodies can also be used, e.g. B. humanized mouse antibodies (Shin et al., 1989; Güssow et Seemann, 1991), bispecific antibodies (Weiner et al., 1993; Goodwin, 1989) or single-chain antibody / toxin fusion proteins (Friedman et al. , 1993). The antibodies can be isolated from sera or from hybridoma supernatants or obtained by recombinant expression as a single-chain antibody (scFv, Johnson et Bird, 1991), complete or fragile tar-like immunoglobulins (Coloma et al., 1992; Nesbit et al. , 1992; Bar bas et al., 1992). The antibodies can be used alone or linked to an agent. If the antibody is to be used therapeutically or for immunoscintigraphy, it can be used with a suitable radioactive isotope e.g. B. ¹³¹I, ¹²⁵I, ¹¹¹In, ¹⁸⁶Re, ⁹⁰Y, 99m Tc or ²¹¹At. This link can cool directly or via a linker mole, e.g. B. a chelating agent. Methods of radiolabeling antibodies are state of the art (Larson et al., 1991; Thomas et al., 1989; Greiner et al., 1993; Sri vastava, 1988; Rhodes et al., 1986). For therapeutic use, the antibody can also be linked to a non-radioactive cytotoxic agent. This can be a cytostatic (Schrappe et al., 1992) or a cytotoxic polypeptide, e.g. B. a bacterial or vegetable toxin, (Vitetta et al., 1991; Kreitman et al., 1993). Such a cytotoxic polypeptide can be covalently, e.g. B. be linked via disulfide bridges with the antibody (Theuer et al., 1993), or in the form of a fusion protein in a einketti gene immunotoxin (Chaudhary et al., 1990; Friedman et al., 1993) with an antibody. The antibody may also be linked to a cytokine or other immunomodulatory polypeptide, e.g. B. with tumor necrosis factor or Interleu kin-2. The antibody can also be linked to an agent which, while not itself cytotoxic, can produce a cytotoxic substance, e.g. B. an enzyme that catalyzes the conversion of an inactive precursor molecule (prodrug) into a cytostatic agent (Wang et al., 1992; Senteretal., 1989).
Die rekombinante Expression von Polypeptiden, die v5-kodierte Sequenzen enthalten, so wie die Herstellung von v5-spezifischen antiidiotypischen Antikörpern können mit Metho den aus dem Stand der Technik durchgeführt werden (Sambrook et al., 1989; Briles et Kearney, 1985).The recombinant expression of polypeptides which contain v5-encoded sequences, see above how the production of v5-specific antiidiotypic antibodies can with metho carried out from the prior art (Sambrook et al., 1989; Briles et Kearney, 1985).
Die erfindungsgemäße Verwendung der Antikörper gegen variante CD44-Epitope kann durch systemische oder topische Applikation erfolgen, beispielsweise durch intravenöse (als Bolus oder Dauerinfusion), intraperitoneale, intramuskuläre, subkutane o. a. Injek tion/Infusion. Es können auch einzelne Organe oder Glieder perfundiert werden. Protokolle für die Verabreichung von konjugierten oder nichtkonjugerten Antikörpern (sei es als kom plette Immunglobuline, Fragmente, rekombinante chimäre Moleküle o. ä.) sind Stand der Technik (Mulshine et al., 1991; Larson et al., 1991; Vitetta et Thorpe, 1991; Vitetta et al., 1991; Breitz et al., 1992; Press et al., 1989; Weiner et al., 1989; Chatal et al., 1989; Sears et al., 1982).The use according to the invention of the antibodies against variant CD44 epitopes can by systemic or topical application, for example by intravenous (as Bolus or continuous infusion), intraperitoneal, intramuscular, subcutaneous, etc. Injec tion / infusion. Individual organs or limbs can also be perfused. Logs for the administration of conjugated or non-conjugated antibodies (whether as a com plette immunoglobulins, fragments, recombinant chimeric molecules or the like) are the state of the art Technology (Mulshine et al., 1991; Larson et al., 1991; Vitetta et Thorpe, 1991; Vitetta et al., 1991; Breitz et al., 1992; Press et al., 1989; Weiner et al., 1989; Chatal et al., 1989; Sears et al., 1982).
Neben der therapeutischen Behandlung von Krebserkrankungen eignen sich die erfindungs gemäßen Mittel zur in-vivo-Diagnostik von Tumoren. Für die Verwendung von mit radio aktiven Isotopen konjugierten Antikörpern zur Immunszintigraphie (Imaging) gibt es eben falls eine Reihe von Protokollen auf deren Grundlage der Fachmann die Erfindung ausfüh ren kann (Siccardi et al., 1989; Keenan et al., 1987; Perkins et Pimm, 1992; Colcher et al., 1987; Thompson et al., 1984).In addition to the therapeutic treatment of cancer, the fiction Appropriate means for the in vivo diagnosis of tumors. For use with radio There are active isotope-conjugated antibodies for immunoscintigraphy (imaging) if a series of protocols on the basis of which the person skilled in the art will carry out the invention ren (Siccardi et al., 1989; Keenan et al., 1987; Perkins et Pimm, 1992; Colcher et al., 1987; Thompson et al., 1984).
Rekombinante Polypeptide, die durch Exon v5 kodierte Aminosäuresequenzen enthalten, sowie v5-spezifische antiidiotypische Antikörper können als Tumorvakzine eingesetzt wer den. Antiidiotypische Antikörper erkennen Epitope innerhalb der variablen Regionen von Immunglobulinen (Idiotope). Unter v5-spezifischen antiidiotypischen Antikörpern sind Antikörper zu verstehen, die gegen Idiotope von Antikörpern gerichtet sind, die v5-kodierte Aminosäuresequenzen erkennen. Setzt man v5-spezifische antiidiotypische Antikörper als Antigene in einem Immunisierungsprotokoll ein, werden gegen diese Antigene Antikörper gebildet, die zum Großteil auch an v5-kodierte Epitope binden. Polypeptide, die v5-kodierte Sequenzen enthalten, und v5-spezifische antiidiotypische Antikörper kann man also zur Immunisierung von Tumorpatienten verwenden. Die Antikörper, die dabei im Organismus des Patienten gegen v5-kodierte Epitope gebildet werden, erkennen Tumorzellen, die diese Epitope exprimieren, und unterstützen so die immunologische Abwehr des Tumors durch den Organismus.Recombinant polypeptides containing amino acid sequences encoded by exon v5 as well as v5-specific anti-idiotypic antibodies can be used as tumor vaccines the. Antiidiotypic antibodies recognize epitopes within the variable regions of Immunoglobulins (idiotopes). Among v5-specific anti-idiotypic antibodies are To understand antibodies directed against idiotopes of antibodies that encoded v5 Recognize amino acid sequences. If one sets v5-specific anti-idiotypic antibodies as Antigens in an immunization protocol are antibodies against these antigens formed, most of which also bind to v5-coded epitopes. Polypeptides that encoded v5 Contain sequences, and v5-specific anti-idiotypic antibodies can be used for Use immunization of tumor patients. The antibodies that are present in the organism of the patient against v5-encoded epitopes, tumor cells recognize these Express epitopes and thus support the immunological defense of the tumor the organism.
Variante CD44-Moleküle sind tumorassoziierte Antigene, die sich sehr gut als Targets für die Immuntherapie und -szintigraphie von Krebserkrankungen eignen. Besonders gut eignen sich dabei variante CD44-Moleküle, die die durch das variable Exon v5 kodierte Aminosäu resequenz, eine allele Variante oder ein Fragment dieser Sequenz enthalten, sowie v5-spezi fische antiidiotypische Antikörper.Variant CD44 molecules are tumor-associated antigens that work very well as targets for immunotherapy and scintigraphy of cancer. Particularly suitable variant CD44 molecules that contain the amino acid encoded by the variable exon v5 contain a sequence, an allelic variant or a fragment of this sequence, and v5-spec fish anti-idiotypic antibodies.
Immunhistochemische Untersuchungen mit monoklonalen Antikörpern gegen verschiedene CD44-Varianten an menschlichem Normal- und Tumorgewebe zeigen, daß viele Tumoren CD44v hoch exprimieren, während dies die entsprechenden Normalgewebe meist nur schwach oder nicht tun.Immunohistochemical studies with monoclonal antibodies against various CD44 variants on human normal and tumor tissue show that many tumors Highly express CD44v, while this usually only corresponds to the corresponding normal tissues weak or not doing.
Die Feinanalyse der Expression verschiedener CD44-Varianten zeigt, daß nicht alle Exons gleichartig in Tumoren überexprimiert sind. Tabelle 1 zeigt eine Untersuchung an Magen tumoren von insgesamt 42 Patienten. In 42/42 Fällen ist eine Expression von varianten CD44-Molekülen nachweisbar (polyklonales Antiserum gegen die Aminosäuresequenzen der Exons v3-v10). Untersuchungen mit Exon-spezifischen monoklonalen Antikörpern zeigt dann, daß Exons v3/v4, v7, v8-v10 nicht und v6 nur in 26/42 Tumoren (62%) exprimiert wird. v5 dagegen wird in 39/42 Tumoren (93%) exprimiert. Im Gegensatz zu v6 lassen sich mit v5-spezifischen Antikörpern sowohl Magentumoren vom diffusen (14/17) als auch vom intestinalen Typ (25/25) erfassen. In Tumoren, die beide Exons exprimieren, ist die Expres sion von v5 meist stärker als die von v6 (Tabelle 2).The fine analysis of the expression of different CD44 variants shows that not all exons are similarly overexpressed in tumors. Table 1 shows an examination on the stomach tumors from a total of 42 patients. In 42/42 cases there is an expression of variants Detectable CD44 molecules (polyclonal antiserum against the amino acid sequences the exons v3-v10). Studies with exon-specific monoclonal antibodies shows then that exons v3 / v4, v7, v8-v10 not and v6 only expressed in 26/42 tumors (62%) becomes. v5, on the other hand, is expressed in 39/42 tumors (93%). In contrast to v6 with v5-specific antibodies both gastric tumors from diffuse (14/17) and from capture intestinal type (25/25). In tumors that express both exons is the expres sion of v5 mostly stronger than that of v6 (Table 2).
Abb. 3 stellt die Ergebnisse einer Studie an 39 kolorektalen Karzinomen dar. Im Gegensatz zu allen anderen Exons wird v5 im Normalgewebe überhaupt nicht, im Tumorgewebe dage gen bereits im frühesten Tumorstadium (frühes Adenom) in über 80% der Tumoren expri miert. v3-Expression wurde überhaupt nicht, v6-Expression erst in späten Tumorstadien deutlich zunehmend und v8-v10 bereits im Normalgewebe exprimiert nachgewiesen. Auch in späteren Stadien der kolorektalen Tumorprogression wird v5 häufiger und in stärkerem Ausmaß exprimiert als v6.Fig. 3 shows the results of a study on 39 colorectal carcinomas. In contrast to all other exons, v5 does not appear at all in normal tissue and in tumor tissue express in the earliest tumor stage (early adenoma) in over 80% of the tumors lubricated. v3 expression was not at all, v6 expression only in late tumor stages significantly increased and v8-v10 already detected in normal tissue. Also in later stages of colorectal tumor progression becomes more common and v5 Extent expressed as v6.
Eine Untersuchung von Brusttumoren (62 invasive Karzinome, 4 in-situ-Karzinome, 9 loka le Rezidive, 16 Lymphknotenmetastasen) ergab ebenfalls, das v5 in diesen Tumoren sehr häufig exprimiert wird (82-100%), häufiger als v3 oder v6 (Tabelle 3).An examination of breast tumors (62 invasive carcinomas, 4 in situ carcinomas, 9 loca le recurrences, 16 lymph node metastases) also showed that v5 was very high in these tumors is expressed frequently (82-100%), more often than v3 or v6 (Table 3).
Für Verfahren der Immuntherapie und -szintigraphie ist es wichtig, daß das tumorassoziierte Antigen, das das Target des Antikörpers darstellt, in Tumor-, nicht jedoch in Normalgewe be exprimiert, andererseits aber in einer möglichst großen Zahl von Tumoren in einem mög lichst frühen Tumorstadium möglichst hoch exprimiert wird, um eine breite und zuverlässige Anwendung zu erlauben. Überraschenderweise erfüllen CD44-Varianten, die durch Exon v5 kodierte Aminosäuresequenzen enthalten, diese Voraussetzungen in hohem Maße. Die er findungsgemäßen Mittel und Verwendungen eignen sich somit hervorragend zur Immun therapie und in-vivo-Diagnostik/Immunszintigraphie von Tumoren, insbesondere von Kar zinomen. For methods of immunotherapy and scintigraphy, it is important that the tumor-associated Antigen, which is the target of the antibody, in tumor but not in normal tissues be expressed, but on the other hand possible in as large a number of tumors as possible As early as possible, the tumor stage is expressed as high as possible in order to achieve a broad and reliable Allow application. Surprisingly, CD44 variants fulfilled by Exon v5 encoded amino acid sequences contain these requirements to a high degree. Which he Agents and uses according to the invention are therefore outstandingly suitable for immune purposes therapy and in vivo diagnostics / immunoscintigraphy of tumors, especially of Kar zinomen.
Fig. 1 Schematische Darstellung einer CD44-Spleißvariante. Diese beispielhafte Variante trägt alle varianten Exonsequenzen an der einzigen Insertionsstelle. Dunkelgraue Kästen symbolisieren CD44-Standardsequenzen (CD44s). Die Lokalisierung der Epitope der monoklonalen Antikörper VFF4, VFF7, VFF8, VFF9, VFF11, VFF14, VFF16 und VFF18 ist durch Balken angezeigt. Alle monoklonalen Antikörper sind exonspezifisch. Fig. 1 Schematic representation of a CD44 splice variant. This exemplary variant carries all variant exon sequences at the single insertion site. Dark gray boxes symbolize CD44 standard sequences (CD44s). The location of the epitopes of the monoclonal antibodies VFF4, VFF7, VFF8, VFF9, VFF11, VFF14, VFF16 and VFF18 is indicated by bars. All monoclonal antibodies are exon-specific.
Fig. 2 Immunhistochemie von normaler Mucosa sowie Adenokarzinomen des Magens. Ei ne fokal betonte anti-CD44v-positive Reaktion zeigt sich in Tumorzellen eines mäßig diffe renzierten Adenokarzinoms (intestinaler Typ nach Lauren) des Magens (a) sowie in einer regionalen Lymphknotenmetastase (b). In normaler Mucosa des Magens mit chronischer Gastritis reagieren die Foci von intestinalen Metaplasien positiv mit mAb VFF4 (c, Pfeile) sowie mit mAb VFF8 (d, Pfeile), begleitet von einer zusätzlichen Reaktion auf der mucoi den Oberfläche und dem foveolären Epithel (d, Pfeilspitzen). Fast alle Becherzellkarzinome des Magens (diffuser Typ nach Lauren) zeigen eine negative Reaktion mit mAb VFF4 (e), und im Gegensatz zu Adenokarzinomen des intestinalen Typs ist das normale mucoide Epi thel negativ (e, Pfeilspitzen). In den meisten Fällen zeigt sich in diesen Becherzellkarzino men eine positive Reaktion mit mAb VFF8 (f), und auch das verbliebene normale mucoide Epithel zeigt Immunreaktivität (f; Pfeilspitzen). (ABC-Methode a, b: anti-CD44v polyklonales Serum, 140 x; c: VFF4, 80x; d: VFF8, 80x; e: VFF4, 210x; f: VFF8, 210x; Gegenfärbung Hämatoxylin). Fig. 2 Immunohistochemistry of normal mucosa and gastric adenocarcinomas. A focal stressed anti-CD44v-positive reaction manifests itself in tumor cells of a moderately differentiated adenocarcinoma (Lauren intestinal type) of the stomach (a) and in a regional lymph node metastasis (b). In normal gastric mucosa with chronic gastritis, the foci of intestinal metaplasias react positively with mAb VFF4 (c, arrows) and with mAb VFF8 (d, arrows), accompanied by an additional reaction on the mucosal surface and the foveolar epithelium (d, Arrowheads). Almost all goblet cell carcinomas of the stomach (Lauren diffuse type) show a negative reaction with mAb VFF4 (e), and in contrast to intestinal type adenocarcinomas, the normal mucoid epithelium is negative (e, arrowheads). In most cases, these goblet cell carcinomas show a positive reaction with mAb VFF8 (f), and the remaining normal mucoid epithelium also shows immunoreactivity (f; arrowheads). (ABC method a, b: anti-CD44v polyclonal serum, 140 x; c: VFF4, 80x; d: VFF8, 80x; e: VFF4, 210x; f: VFF8, 210x; counterstaining hematoxylin).
Fig. 3 Expression varianter CD44-Exons in verschiedenen Stadien der kolorektalen Tu morprogression. Ergebnisse aus immunhistochemischen Anfärbungen von Gewebsschnitten (Beispiel 2). Dunkelgraue Balken zeigen den Prozentsatz positiver Tumoren an. Hellgraue Balken zeigen Proben mit nur fokaler Anfärbung an. Fig. 3 Expression of variant CD44 exon morprogression in various stages of colorectal Tu. Results from immunohistochemical staining of tissue sections (Example 2). Dark gray bars indicate the percentage of positive tumors. Light gray bars show samples with only focal staining.
Kolon: Normale und pathologische Gewebe wurden aus den Beständen der Abteilung für Pathologie, Academic Medical Center, Universität Amsterdam, Niederlande, entnommen.Colon: Normal and pathological tissues were taken from the department's stocks Pathology, taken from Academic Medical Center, University of Amsterdam, The Netherlands.
Kolorektale Karzinome (n=39) wurden nach der Klassifikation von Dukes (1937, 1980) in Stadien eingeteilt, in Dukes A (n=9), Krankheit beschränkt auf die Darmwand; Dukes B (n=14), Ausdehnung über die Muskelschicht hinaus ohne Metastasierung; Dukes C/D (n=16), Tumoren mit regionalen bzw. Fernmetastasen. Adenome wurden unterteilt in frühe Adenome (Durchmesser (1 cm, n=11) und späte Adenome (Durchmesser) 1 cm, n=12) und wurden als niedrig oder hoch differenziert nach Standardkriterien gradiert.Colorectal carcinomas (n = 39) were classified according to the Dukes (1937, 1980) classification Stages divided into dukes A (n = 9), disease limited to the intestinal wall; Dukes B (n = 14), expansion beyond the muscle layer without metastasis; Dukes C / D (n = 16), tumors with regional or distant metastases. Adenomas were divided into early ones Adenomas (diameter (1 cm, n = 11) and late adenomas (diameter) 1 cm, n = 12) and were graded as low or high differentiated according to standard criteria.
Magen: Tumorproben und Normalgewebe wurden aus den Beständen der Abteilung Patho logie der Universität Würzburg, Deutschland, ausgewählt. Die Proben waren unmittelbar nach der chirurgischen Entnahme schockgefroren und bis zur Verwendung bei -80°C gela gert worden. Normalgewebe wurde von zwölf verschiedenen Tumor-Patienten sowohl aus der Korpus- als auch aus der Antrumregion des Magens entnommen. Pathologische Gewebe wurden von einer Gesamtzahl von 47 Patienten mit einem Durchschnittsalter von 63 Jahren erhalten. Von den Primärkarzinomen gehörten 29 zum intestinalen und 18 zum diffusen Typ nach Lauren (1965). Die Tumorstadien reichten von lokalisiert (pT1) bis ausgedehnt (pT4), die histologische Gradierung von gut differenzierten (G1) bis schlecht differenzierten (G3) Adenokarzinomen.Stomach: Tumor samples and normal tissues were taken from the Patho department of the University of Würzburg, Germany. The samples were immediate shock-frozen after surgical removal and gela at -80 ° C until use been tied. Normal tissue was made up of twelve different tumor patients both the body as well as the antrum region of the stomach. Pathological tissue were taken from a total of 47 patients with an average age of 63 years receive. Of the primary cancers, 29 were intestinal and 18 were diffuse after Lauren (1965). The tumor stages ranged from localized (pT1) to extensive (pT4), histological grading from well differentiated (G1) to poorly differentiated (G3) Adenocarcinomas.
Mamma: Gefrorene Gewebe (aufbewahrt bei -70°C) wurden von der Universitätsfrauenkli nik Heidelberg, Deutschland, erhalten. Eingeschlossen waren 62 Proben von primären Mammakarzinomen, 9 lokale Rezidive von Mammakarzinomen, 4 Fälle von reinen in-situ- Karzinomen und 16 axilläre Lymphknotenmetastasen (von den gleichen Patienten wie die Primärtumoren). Die Fälle wurden zufällig ausgewählt und schlossen eine repräsentative Auswahl von histologischen Tumortypen, Stadien und Gradierungen ein. Zum Vergleich wurden Proben von normalem Brustgewebe, duktalen Hyperplasien und Fibroadenomen ausgewählt. Mamma: Frozen tissues (stored at -70 ° C) were obtained from the University Women's Hospital nik Heidelberg, Germany. 62 samples from primary were included Breast cancer, 9 local recurrences of breast cancer, 4 cases of pure in-situ Carcinomas and 16 axillary lymph node metastases (from the same patient as that Primary tumors). The cases were chosen at random and included a representative one Selection of histological tumor types, stages and gradings. For comparison were samples of normal breast tissue, ductal hyperplasia and fibroadenomas selected.
Die gesamte variante Region des HPKII-Typs von CD44v (Hofmann et al., 1991) wurde aus menschlicher Keratinozyten-cDNA durch Polymerase-Kettenreaktion (PCR) amplifi ziert. Die beiden PCR-Primer 5′-CAGGCTGGGAGCCAAATGAAGAAAATG-3′, Posi tionen 25-52, und 5′-TGATAAGGAACGATTGACATTAGAGTTGGA-3′, Positionen 1013-984 der LCLC97-varianten Region, wie von Hofmann et al. beschrieben, enthielten eine EcoRI-Erkennungsstelle, die benutzt wurde, um das PCR-Produkt direkt in den Vektor pGEX-2T (Smith et al., 1988) zu klonieren. Das resultierende Konstrukt (pGEX CD44v HPKII, v3-v10) kodiert für ein Fusionsprotein von 70 kD.The entire variant region of the HPKII type of CD44v (Hofmann et al., 1991) was amplify from human keratinocyte cDNA by polymerase chain reaction (PCR) graces. The two PCR primers 5'-CAGGCTGGGAGCCAAATGAAGAAAATG-3 ', Posi tions 25-52, and 5′-TGATAAGGAACGATTGACATTAGAGTTGGA-3 ′, positions 1013-984 of the LCLC97 variant region as described by Hofmann et al. described an EcoRI recognition site that was used to insert the PCR product directly into the vector to clone pGEX-2T (Smith et al., 1988). The resulting construct (pGEX CD44v HPKII, v3-v10) codes for a fusion protein of 70 kD.
Um Subklone der varianten Regionen zu erhalten, die für Affinitätsreinigungen und Western-Blot-Analysen verwendet werden konnten, wurden Fragmente kloniert, die DI (v3), DII/III (v5, v6), und DIII (v6, v7) enthielten, wobei die passenden Restriktionsschnitt stellen verwendet wurden. Fusionsprotein DI enthält die CD44-Sequenz, die von Stamen kovic et al. (1989) beschrieben wurde, von Position 744 bis zur Position 142 der Sequenz von variantem CD44, wie sie von Hofmann et al. (1991) beschrieben wurde. Fusionsprotein DII/III enthält die variante Sequenz von Position 290-460, Fusionsprotein DIII die variante Sequenz von Position 378-638 (Hofmann et al., 1991). Die DI und DIII enthaltenden Fragmente wurden in das pGEX-Vektorsystem, das DII/III-Fragment in den pATH-Vektor (Angel et al., 1988) kloniert.In order to obtain subclones of the variant regions which are used for affinity purifications and Western blot analyzes could be used, fragments were cloned that DI (v3), DII / III (v5, v6), and DIII (v6, v7) included, with the appropriate restriction cut places were used. Fusion protein DI contains the CD44 sequence derived from stamen kovic et al. (1989), from position 744 to position 142 of the sequence of variant CD44, as described by Hofmann et al. (1991). Fusion protein DII / III contains the variant sequence from position 290-460, fusion protein DIII the variant Sequence from position 378-638 (Hofmann et al., 1991). The DI and DIII containing Fragments were in the pGEX vector system, the DII / III fragment in the pATH vector (Angel et al., 1988).
Die Herstellung und Reinigung des polyklonalen Antiserums gegen die variante Region des CD44-Moleküls ist in der Literatur beschrieben (Heider et al., 1993).The production and purification of the polyclonal antiserum against the variant region of the CD44 molecule is described in the literature (Heider et al., 1993).
Weibliche BALB/c-Mäuse wurden mit affinitätsgereinigtem Fusionsprotein immunisiert, das aus pGEX CD44v HPKJI (Exons v3-v10) wie oben beschrieben erhalten wurde. Milzzellen eine Tieres mit hohem Antikörpertiter wurde mit P3X63Ag8.653-Myelomzellen unter Ver wendung von Polyethylenglykol 4000 fusioniert. Hybridome wurden in HAT-Medium se lektiert (Kearney et al., 1979). Bestimmung der Antikörpertiter im Serum sowie das An tikörperscreening wurden mittels ELISA durchgeführt. Die Microtiterplatten wurden mit Fusionsprotein beschichtet, mit seriellen Verdünnungen von Serumproben oder Hybridoma überständen inkubiert, und spezifische Antikörper wurden mit Peroxidase-gekoppelten An tikörpern gegen Maus-IgG detektiert. Hybridome, die mit Glutathion-Transferase reagier ten, wurden eliminiert. Die verbleibenden Antikörper wurden mit ELISA-Tests weiter cha rakterisiert, wobei Fusionsproteine der variablen Domänen DI (Exon v3), DII/III (Exons v5, v6),. DIII (Exons v6, v7), DI-IV (Exons v3-v8), DIII-VI (Exons v7-v10) bzw. v6 (Exon v6) [v5 v6 v7] verwendet wurden. Die Reaktivität der Antikörper mit menschlichen Hautkeratinozyten wurde immunhistochemisch untersucht.Female BALB / c mice were immunized with affinity-purified fusion protein that from pGEX CD44v HPKJI (exons v3-v10) as described above. Spleen cells an animal with a high antibody titer was treated with P3X63Ag8.653 myeloma cells under Ver using polyethylene glycol 4000 fused. Hybridomas were se in HAT medium read (Kearney et al., 1979). Determination of the antibody titer in the serum and the An antibody screening was performed using ELISA. The microtiter plates were with Fusion protein coated, with serial dilutions of serum samples or hybridoma supernatants were incubated, and specific antibodies were coupled with peroxidase-coupled An antibodies against mouse IgG detected. Hybridomas that react with glutathione transferase were eliminated. The remaining antibodies were further cha with ELISA tests characterized, fusion proteins of the variable domains DI (exon v3), DII / III (exons v5, v6) ,. DIII (exons v6, v7), DI-IV (exons v3-v8), DIII-VI (exons v7-v10) and v6 (Exon v6) [v5 v6 v7] were used. The reactivity of the antibodies with human Skin keratinocytes were examined immunohistochemically.
Die Exonspezifität verschiedener verwendeter monoklonaler Antikörper (VFF4, VFF7, VFF8, VFF9, VFF11, VFF14, VFF16 und VFF18) ist in Fig. 1 dargestellt.The exon specificity of various monoclonal antibodies used (VFF4, VFF7, VFF8, VFF9, VFF11, VFF14, VFF16 and VFF18) is shown in FIG. 1.
Gefrierschnitte wurden in eisgekühltem Methanol 10 min. fixiert, in PBS (8 g/l NaCl, 0.2 g/l KCl, 1.44 g/l Na₂HPO₄, 0.24 g/l KH₂PO₄, pH 7.4) gewaschen und mit normalem Ziegen serum (10% in PBS) präinkubiert. Dann wurden sie 3× mit PBS gewaschen und für 1 Stun de mit dem Primärantikörper (in PBS, 1% BSA) inkubiert. Endogene Peroxidase wurde mit 0.3% H₂O₂ in Methanol blockiert und die Schnitte mit biotinyliertem Zweitantikörper (entweder anti-Maus oder anti-Kaninchen F(ab′)₂, DAKO Corp., Santa Barbara, CA, USA, abhängig vom verwendeten Primärantikörper) inkubiert. Der Immunkomplex wurde mit Meerrettich-Peroxidase visualisiert, die als Streptavidin-Biotin-Peroxidasekomplex an Bio tin gekoppelt wurde (DAKO). Nach dreißigminütiger Inkubation mit dem Streptavidin-Bio tin-Peroxidase-Komplex wurden die Schnitte mit 3,3-Amino-9-ethyl-carbazol (Sigma Che micals, Deisenhofen, Deutschland) für 5 bis 10 min. entwickelt und die Reaktion mit H₂O abgestoppt. Die Zellen wurden mit Hämatoxylin gegengefärbt, mit Glyzerin-Gelatine einge deckelt und mikroskopisch untersucht.Frozen sections were frozen in ice-cold methanol for 10 min. fixed, in PBS (8 g / l NaCl, 0.2 g / l KCl, 1.44 g / l Na₂HPO₄, 0.24 g / l KH₂PO₄, pH 7.4) washed and with normal goat serum (10% in PBS) preincubated. Then they were washed 3 times with PBS and for 1 hour de incubated with the primary antibody (in PBS, 1% BSA). Endogenous peroxidase was identified with 0.3% H₂O₂ blocked in methanol and the sections with biotinylated second antibody (either anti-mouse or anti-rabbit F (ab ′) ₂, DAKO Corp., Santa Barbara, CA, USA, depending on the primary antibody used). The immune complex was created with Horseradish peroxidase visualized as a streptavidin-biotin-peroxidase complex at Bio tin was coupled (DAKO). After 30 minutes of incubation with the streptavidin bio The tin-peroxidase complex was cut with 3,3-amino-9-ethyl-carbazole (Sigma Che micals, Deisenhofen, Germany) for 5 to 10 min. developed and the reaction with H₂O stopped. The cells were counterstained with hematoxylin and gelatinized with glycerin lids and examined microscopically.
Tumoren wurden als "positiv" bezeichnet, wenn mehr als 10% der Tumorzellen angefärbt waren. Waren weniger als 10% der Tumorzellen gefärbt, wurde dies als "fokal" bezeichnet. Tumors were said to be "positive" when more than 10% of the tumor cells stained were. If less than 10% of the tumor cells were stained, this was referred to as "focal".
Angel P, Allegretto E A, Okuio S T, Hatton K, Boyle W J, Hunter T, Karin M. Oncogene jun encodes a sequence-specific trans-activator similar to AP-1. Nature 332, 166 (1988).Angel P, Allegretto E A, Okuio S T, Hatton K, Boyle W J, Hunter T, Karin M. Oncogene jun encodes a sequence-specific trans-activator similar to AP-1. Nature 332, 166 (1988).
Barbas C F, Björling E, Chiodi F, Dunlop N, Cababa D, Jones T M, Zebedee S L, Persson M A A, Nara P L, Norrby E, Burton D R. Recombinant human Fab fragments neutralize human type 1 immunodeficiency virus in vifro. Proc. Natl. Acad. Sci. US.A. 89, 9339-9343 (1992).Barbas C F, Björling E, Chiodi F, Dunlop N, Cababa D, Jones T M, Zebedee S L, Persson M A A, Nara P L, Norrby E, Burton D R. Recombinant human Fab neutralize fragments human type 1 immunodeficiency virus in vifro. Proc. Natl. Acad. Sci. USA. 89, 9339-9343 (1992).
Brasch R C. New directions in the development of MR imaging contrast media. Radiology 183, 1-11 (1992).Brasch R C. New directions in the development of MR imaging contrast media. Radiology 183: 1-11 (1992).
Breitz H B, Weiden P L, Vanderheyden J-L, Appelbaum J W, Bjorn M J, Fer M F, Wolf S B, Ratcliff B A, Seiler C A, Foisie D C, Fisher D R, Schroff R W, Fritzberg A R, Abrams P G. Clinical experience with rhenium-186-labeled monoclonal antibodies for radioim munotherapy: results of phase I trials. J Nucl. Med. 33, 1099-1112 (1992).Breitz H B, Weiden P L, Vanderheyden J-L, Appelbaum J W, Bjorn M J, Fer M F, Wolf S B, Ratcliff B A, Seiler C A, Foisie D C, Fisher D R, Schroff R W, Fritzberg A R, Abrams P G. Clinical experience with rhenium-186-labeled monoclonal antibodies for radioim munotherapy: results of phase I trials. J Nucl. Med. 33: 1099-1112 (1992).
Briles D E, Kearney J F. Antiidiotypic antibodies. MethoasEnzymol. 116, 174-189 (1985).Briles D E, Kearney J F. Antiidiotypic antibodies. MethoasEnzymol. 116: 174-189 (1985).
Chatal J-F, Saccavini J-C, Gestin J-F, Th´drez P, Curtet C, Kremer M, Guerreau D, Nolib´ D, Fumoleau P, Guillard Y. Biodistribution of indium-111-labeled OC 125 monoclonal an tibody intraperitoneally injected into patients operated on for ovarian carcinomas. Cancer Res. 49, 3087-3094 (1989).Chatal J-F, Saccavini J-C, Gestin J-F, Th´drez P, Curtet C, Kremer M, Guerreau D, Nolib´ D, Fumoleau P, Guillard Y. Biodistribution of indium-111-labeled OC 125 monoclonal tibody intraperitoneally injected into patients operated on for ovarian carcinomas. Cancer Res. 49, 3087-3094 (1989).
Chaudhary V K, Batra J K, Galdo M G, Willingham M C, Fitzgerald D J, Pastan I. A rapid method of cloning functional variable-region antibody genes in Escherichia coli as single chain immunotoxins. Proc. Natl. Acad. Sci. U.S.A. 87, 1066 (1990).Chaudhary V K, Batra J K, Galdo M G, Willingham M C, Fitzgerald D J, Pastan I. A rapid method of cloning functional variable-region antibody genes in Escherichia coli as single chain immunotoxins. Proc. Natl. Acad. Sci. U.S.A. 87, 1066 (1990).
Colcher D, Esteban J, Carrasquillo J A, Sugarbaker P, Reynolds J C, Bryant G, Larson S M, Schlom J. Complementation of intracavitary and intravenous administration of a mono clonal antibody (B72.3) in patients with carcinoma. Cancer Res. 47, 4218-4224 (1987).Colcher D, Esteban J, Carrasquillo J A, Sugarbaker P, Reynolds J C, Bryant G, Larson S M, Schlom J. Complementation of intracavitary and intravenous administration of a mono clonal antibody (B72.3) in patients with carcinoma. Cancer Res. 47, 4218-4224 (1987).
Coloma M J, Hastings A, Wims L A, Morrison S L. Novel vectors for the expression of antibody molecules using variable regions generated by polymerase chain reaction. J. Im munol. Methods 152, 89-104 (1992).Coloma M J, Hastings A, Wims L A, Morrison S L. Novel vectors for the expression of antibody molecules using variable regions generated by polymerase chain reaction. J. Im munol. Methods 152: 89-104 (1992).
Dukes, C. E. Histological grading of rectal cancer. Proc. R. Soc. Med 30, 371-376 (1937)
Dukes, C. E. The classification of cancer of the rectum. Dis. Colon Rectum 23, 605-611
(1980).Dukes, CE Histological grading of rectal cancer. Proc. R. Soc. Med 30, 371-376 (1937)
Dukes, CE The classification of cancer of the rectum. Dis. Colon Rectum 23, 605-611 (1980).
Friedmann P N, McAndrew S J, Gawlak S L, Chace D, Trail P A, Brown J P, Siegall C B.Friedmann P N, McAndrew S J, Gawlak S L, Chace D, Trail P A, Brown J P, Siegall C B.
BR96 sFv-PE40, a potent single-chain immunotoxin that selectively kills carcinoma cells. Cancer Res. 53, 334-339 (1993). BR96 sFv-PE40, a potent single-chain immunotoxin that selectively kills carcinoma cells. Cancer Res. 53, 334-339 (1993).
Fritzberg A R, Abrams P G, Beaumier P L. Specific and stable labeling of antibodies with technetium-99m with a diamide dithiolate chelating agent. Proc. Natl. Acad. Sci. U.S.A. 85, 4025 (1988).Fritzberg A R, Abrams P G, Beaumier P L. Specific and stable labeling of antibodies with technetium-99m with a diamond dithiolate chelating agent. Proc. Natl. Acad. Sci. U.S.A. 85, 4025 (1988).
Goodwin D A. A new appoach to the problem of targeting specific monoclonal antibodies to human tumors using anti-hapten chimeric antibodies. J Nucl. Med. Biol. 16, 645 (1989).Goodwin DA A new appoach to the problem of targeting specific monoclonal antibodies to human tumors using anti-hapten chimeric antibodies. J Nucl. Med. Biol. 16, 645 (1989).
Greiner J W, Ullmann C D, Nieroda C, Qi C-F, Eggensperger D, Shimada S, Steinberg S M, Schlom J. Improved radioimmunotherapeutic efficacy of an anticarcinoma monoclonal antibody (¹³¹I-CC49) when given in combination with γ-interferon. Cancer Res. 53, 600- 608 (1993).Greiner J W, Ullmann C D, Nieroda C, Qi C-F, Eggensperger D, Shimada S, Steinberg S M, Schlom J. Improved radioimmunotherapeutic efficacy of an anticarcinoma monoclonal antibody (131 I-CC49) when given in combination with γ-interferon. Cancer Res. 53, 600- 608 (1993).
Günthert U, Hofmann M, Rudy W, Reber S, Zöller M, Haußmann I, Matzku S, Wenzel A, Ponta H, and Herrlich P. A new variant of glycoprotein CD44 confers metastatic potential to rat carcinoma cells. Cell 65, 13-24 (1991).Günthert U, Hofmann M, Rudy W, Reber S, Zöller M, Haußmann I, Matzku S, Wenzel A, Ponta H, and Herrlich P. A new variant of glycoprotein CD44 confers metastatic potential to rat carcinoma cells. Cell 65, 13-24 (1991).
Güssow D, Seemann G. Humanization of monoclonal antibodies. Methods Enzymol. 203, 99-121 (1991):Güssow D, Seemann G. Humanization of monoclonal antibodies. Methods Enzymol. 203, 99-121 (1991):
Heider K-H, Hofmann M, Horst E, van den Berg F, Ponta H, Herrlich P, and Pals S T. A human homologue of the rat metastasis-associated variant of CD44 is expressed in colo rectal carcinomas and adenomatous polyps. J Cell Biol. 120, 227-233 (1993).Heider K-H, Hofmann M, Horst E, van den Berg F, Ponta H, Herrlich P, and Pals S T. A human homologue of the rat metastasis-associated variant of CD44 is expressed in colo rectal carcinomas and adenomatous polyps. J Cell Biol. 120, 227-233 (1993).
Hider R C, Hall A D. Clinically useful chelators of tripositive elements. Prog. Med. Chem. 28, 41-173 (1991).Hider R C, Hall A D. Clinically useful chelators of tripositive elements. Prog. Med. Chem. 28: 41-173 (1991).
Hofmann M, Rudy W, Zöfler M, Tölg C, Ponta H, Herrlich P, and Günthert U. CD44 splice variants confer metastatic behavior in rats: homologous sequences are expressed in human tumor cell lines. Cancer Res. 51, 5292-5297 (1991).Hofmann M, Rudy W, Zöfler M, Tölg C, Ponta H, Herrlich P, and Günthert U. CD44 splice variants confer metastatic behavior in rats: homologous sequences are expressed in human tumor cell lines. Cancer Res. 51, 5292-5297 (1991).
Johnson S, Bird R E. Construction of single-chain derivatives of monoclonal antibodies and their production in Escherichia coli. Methods Enzymol. 203, 88-98 (1991).Johnson S, Bird R E. Construction of single-chain derivatives of monoclonal antibodies and their production in Escherichia coli. Methods Enzymol. 203: 88-98 (1991).
Kearney J F, Radbruch A, Liesegang B, Rajewski K. A new mouse myeloma cell line that has lost imunoglobulin expression but permits construction of antibody-secreting hybrid cell lines. J. Immunol. 123, 1548 (1979).Kearney J F, Radbruch A, Liesegang B, Rajewski K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits construction of antibody-secreting hybrid cell lines. J. Immunol. 123, 1548 (1979).
Keenan A M, Weinstein J N, Carrasquillo J A, Bunn P A, Reynolds J C, Foon K A et al. Immunolymphoscintigraphy and the dose dependence of ¹¹¹In-labeled T101 monoclonal an tibody in patients with cutaneous T-cell lymphoma. Cancer Res. 47, 6093-6099 (1987).Keenan A M, Weinstein J N, Carrasquillo J A, Bunn P A, Reynolds J C, Foon K A et al. Immunolymphoscintigraphy and the dose dependence of ¹¹¹In-labeled T101 monoclonal tibody in patients with cutaneous T-cell lymphoma. Cancer Res. 47, 6093-6099 (1987).
Köhler G, Milstein C: Continous cultures of fused cell secreting antibody of predefined specificity. Nature 256, 495-497 (1975).Köhler G, Milstein C: Continuous cultures of fused cell secreting antibody of predefined specificity. Nature 256: 495-497 (1975).
Koopman G, Heider K-H, Horts E, Adolf G R, van den Berg F, Ponta H, Herrlich P, Pals S T. Activated human lymphocytes and aggressive Non-Hodgkin′s lymphomas express a ho mologue of the rat metastasis-associated variant of CD44. J Exp. Med. 177, 897-904 (1993).Koopman G, Heider K-H, Horts E, Adolf G R, van den Berg F, Ponta H, Herrlich P, Pals S T. Activated human lymphocytes and aggressive non-Hodgkin's lymphomas express a ho mologue of the rat metastasis-associated variant of CD44. J Exp. Med. 177, 897-904 (1993).
Kreitman R J Hansen H J, Jones A L, Fitzgerald D J P, Goldenberg D M, Pastan I. Pseu domonas exotoxin-based immunotoxins containing the antibody LL2 or LL2-Fab′ induce regression of subcutaneous human B-cell lymphoma in mice. Cancer Res. 53, 819-825 (1993).Kreitman R J Hansen H J, Jones A L, Fitzgerald D J P, Goldenberg D M, Pastan I. Pseu domonas exotoxin-based immunotoxins containing the antibody LL2 or LL2-Fab ′ induce regression of subcutaneous human B-cell lymphoma in mice. Cancer Res. 53, 819-825 (1993).
Larson S M, Cheung N-K V, Leibel S A. Radioisotope Conjugates. In: DeVita V T, Hell man S, Rosenberg S A (Hrsg.). Biologic therapy of cancer. J. B. Lippincott Comp., Phila delphia, 496-511 (1991).Larson S M, Cheung N-K V, Leibel S A. Radioisotope Conjugates. In: DeVita V T, Hell man S, Rosenberg S A (ed.). Biologic therapy of cancer. J. B. Lippincott Comp., Phila delphia, 496-511 (1991).
Lauren P. The two histological main types of gastric carcinoma: diffuse and so-called inte stinal-type carcinoma. Actapath. microbiol. scand. 64, 31-49 (1965).Lauren P. The two histological main types of gastric carcinoma: diffuse and so-called inte stinal-type carcinoma. Actapath. microbiol. scand. 64, 31-49 (1965).
Mulshine J L, Magnani J L, Linnoila R I: Applications of monoclonal antibodies in the treatment of solid tumors. In: DeVita V T, Hellman S, Rosenberg S A (Hrsg.). Biologic therapy of cancer. J. B. Lippincott Comp., Philadelphia, 563-588 (1991).Mulshine J L, Magnani J L, Linnoila R I: Applications of monoclonal antibodies in the treatment of solid tumors. In: DeVita V T, Hellman S, Rosenberg S A (ed.). Biologic therapy of cancer. J. B. Lippincott Comp., Philadelphia, 563-588 (1991).
Nesbit M, Fu Z F, McDonald-Smith J, Steplewski Z, Curtis P J. Production of a functional monoclonal antibody recognizing human colorectal carcinoma cells from a baculovirus ex pression system. J Immunol. Methods 151, 201-208 (1992).Nesbit M, Fu Z F, McDonald-Smith J, Steplewski Z, Curtis P J. Production of a functional monoclonal antibody recognizing human colorectal carcinoma cells from a baculovirus ex pression system. J Immunol. Methods 151, 201-208 (1992).
Perkins A C, Pimm M V. A role for gamma scintigraphy in cancer immunology and immun otherapy. Eur. J Nucl. Med. 19, 1054-1063 (1992).Perkins A C, Pimm M V. A role for gamma scintigraphy in cancer immunology and immun otherapy. Eur. J Nucl. Med. 19, 1054-1063 (1992).
Press O W, Eary J F, Badger C C, Martin P J, Appelbaum F R, Levy R, Miller R, Brown S, Nelp W B, Krohn K A, Fisher D, DeSantes K, Porter B, Kidd P, Thomas E D, Bernstein I D. Treatment of reftactory Non-Hodgkin′s lymphoma with radiolabeled MB-1 (anti-CD37) antibody. J Clin. Oncol. 7, 1027-1038 (1989).Press O W, Eary J F, Badger C C, Martin P J, Appelbaum F R, Levy R, Miller R, Brown S, Nelp W B, Krohn K A, Fisher D, DeSantes K, Porter B, Kidd P, Thomas E D, Bernstein I D. Treatment of reftactory Non-Hodgkin's lymphoma with radiolabeled MB-1 (anti-CD37) antibody. J Clin. Oncol. 7: 1027-1038 (1989).
Rhodes B A, Zamora P O, Newell K D. Technetium-99m labeling of murine monoclonal antibody fragments. J: Nucl. Med. 27, 685 (1986).Rhodes B A, Zamora P O, Newell K D. Technetium-99m labeling of murine monoclonal antibody fragments. J: Nucl. Med. 27, 685 (1986).
Sambrook J, Fritsch E E, Maniatis I. Molecular cloning, Cold Spring Harbor Laboratory Press, Cold Spring Harbor (1989).Sambrook J, Fritsch E E, Maniatis I. Molecular cloning, Cold Spring Harbor Laboratory Press, Cold Spring Harbor (1989).
Saccavini J C, Curtet C, Bohy J, Tellier C, Bourgoin C, Lhoste J M. Magnetic resonance imaging studies on nude mice grafted with colorectal adenocarcinoma using gadolinium labeled monoclonal antibody. Invest. Radiol. 23, Suppl. 1, 292-293 (1988).Saccavini J C, Curtet C, Bohy J, Tellier C, Bourgoin C, Lhoste J M. Magnetic resonance imaging studies on nude mice grafted with colorectal adenocarcinoma using gadolinium labeled monoclonal antibody. Invest. Radiol. 23, Suppl. 1, 292-293 (1988).
Schrappe M, Bumol T F, Apelgren L D, Briggs S L, Koppel G A, Markowitz D D, Mueller B M, Reisfeld R A. Long-term growth suppression of human glioma xenografts by chemo immunoconjugates of 4-desacetylvinblastine-3 -carboxyhydrazide and monoclonal antibody 9.2.27. Cancer Res. 52, 3838-3844 (1992). Schrappe M, Bumol T F, Apelgren L D, Briggs S L, Koppel G A, Markowitz D D, Mueller B M, paddy field R A. Long-term growth suppression of human glioma xenografts by chemo immunoconjugates of 4-desacetylvinblastine-3-carboxyhydrazide and monoclonal antibody 9.2.27. Cancer Res. 52, 3838-3844 (1992).
Screaton G R, Bell M V, Jackson D G, Cornelis F, Gerth U, and Beil J I. Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons. Proc. Natl. Acad. Sci. U.S.A. 89, 12160-12164 (1992).Screaton G R, Bell M V, Jackson D G, Cornelis F, Gerth U, and Beil J I. Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons. Proc. Natl. Acad. Sci. U.S.A. 89, 12160-12164 (1992).
Sears H F, Mattis J, Herlyn D, Häyry P, Atkinson B, Ernst C, Steplewski Z, Koprowski H. Phase-I clinical trial of monoclonal antibody in treatment of gastrointestinal tumours. Lancet 1982 (1), 762-765 (1982).Sears H F, Mattis J, Herlyn D, Häyry P, Atkinson B, Ernst C, Steplewski Z, Koprowski H. Phase-I clinical trial of monoclonal antibody in treatment of gastrointestinal tumors. Lancet 1982 (1), 762-765 (1982).
Senter P D, Schreiber G J, Hirschberg D L, Ashe S A, Hellström K E, Hellström I. En hancement of the in vitro and in vivo antitumor activities of phosphoiylated mitomycin C and etoposide derivatives by monoclonal antibody-alkaline phosphatase conjugates. Cancer Res. 49, 5789-5792 (1989).Senter P D, Schreiber G J, Hirschberg D L, Ashe S A, Hellström K E, Hellström I. En enhancement of the in vitro and in vivo antitumor activities of phosphoiylated mitomycin C and etoposide derivatives by monoclonal antibody-alkaline phosphatase conjugates. Cancer Res. 49, 5789-5792 (1989).
Shin S-U, Morrison S L. Production and properties of chimeric antibody molecules. Methods Enzymol. 178, 459-476 (1989).Shin S-U, Morrison S L. Production and properties of chimeric antibody molecules. Methods Enzymol. 178: 459-476 (1989).
Siccardi A G, Buraggi G L, Callegaro L, Colella A C, DeFilippi P G et al. Immunoscinti graphy of adenocarcinomas by means of radiolabeled F(ab′)₂ fragments of an anti-carcino embryonic antigen monoclonal antibody: a multicenter study. Cancer Res. 49, 3095-3103 (1989).Siccardi A G, Buraggi G L, Callegaro L, Colella A C, DeFilippi P G et al. Immunoscinti graph of adenocarcinomas by means of radiolabeled F (ab ′) ₂ fragments of an anti-carcino embryonic antigen monoclonal antibody: a multicenter study. Cancer Res. 49, 3095-3103 (1989).
Smith D B, Johnson K S. Single-step purification of polypetides expressed in Eseherichia coli as fusions with glutathione S-transferase. Gene 67, 31 (1988).Smith D B, Johnson K S. Single-step purification of polypetides expressed in Eseherichia coli as fusions with glutathione S-transferase. Gene 67, 31 (1988).
Srivastava S C (Hrsg). Radiolabeled monoclonal antibodies for imaging and therapy. Lift Sciences Series A 152, Plenum New York (1988).Srivastava S C (ed.). Radiolabeled monoclonal antibodies for imaging and therapy. lift Sciences Series A 152, New York Plenum (1988).
Stamenkovic I, Amiot M, Pesando J M, and Seed B. A lymphocyte molecule implicated in lymph node homing is a member of the cartilage link protein family. Cell 56, 1057-1062 (1989).Stamenkovic I, Amiot M, Pesando J M, and Seed B. A lymphocyte molecule implicated in lymph node homing is a member of the cartilage link protein family. Cell 56, 1057-1062 (1989).
Theuer C P, Kreitman R J, Fitzgerald D J, Pastan I. Immunotoxins made with a recombi nant form of pseudomonas exotoxin A that do not require proteolysis for activity. Cancer Res. 53, 340-347 (1993).Theuer C P, Kreitman R J, Fitzgerald D J, Pastan I. Immunotoxins made with a recombi nant form of pseudomonas exotoxin A that do not require proteolysis for activity. Cancer Res. 53, 340-347 (1993).
Thomas G D, Dykes P W, Bradwell A R. Antibodies for tumour immunodetection and methods for antibody radiolabeling. In: Catty D (Hrsg.). Antibodies. IRL Press Oxford, 223-244 (1989).Thomas G D, Dykes P W, Bradwell A R. Antibodies for tumor immunodetection and methods for antibody radiolabeling. In: Catty D (ed.). Antibodies. IRL Press Oxford, 223-244 (1989).
Thompson C H, Stacker S A, Salehi N, Lichtenstein M, Leyden M J, Andrews J T. Immu noscintigraphy for detection of lymph node metastases from breast cancer. Lancet 1984 (2), 1245-1247 (1984).Thompson C H, Stacker S A, Salehi N, Lichtenstein M, Leyden M J, Andrews J T. Immu noscintigraphy for detection of lymph node metastases from breast cancer. Lancet 1984 (2), 1245-1247 (1984).
Tölg C, Hofmann M, Herrlich P, Ponta H. Splicing choice from ten variant exons establishes CD44 variability. Nueleic Acids Res. 21, 1225-1229 (1993). Tölg C, Hofmann M, Herrlich P, Ponta H. Splicing choice from ten variant exons establishes CD44 variability. Nueleic Acids Res. 21, 1225-1229 (1993).
Vitetta E S, Thorpe P E. Immunotoxins. In: DeVita V T, Hellman S, Rosenberg S A (Hrsg.). Biologie therapy of cancer. J. B. Lippincott Comp., Philadelphia, 482-495 (1991)Vitetta E S, Thorpe P E. Immunotoxins. In: DeVita V T, Hellman S, Rosenberg S A (Ed.). Biology therapy of cancer. J. B. Lippincott Comp., Philadelphia, 482-495 (1991)
Vitetta E S, Stone M, Amiot P, Fay J, May R, Till M, Newman J, Clark P, Collins R, Cun ningham D, Ghetie V, Uhr J W, Thorpe P E. Phase I immunotoxin trial in patients with B- cell lymphoma. Cancer Res. 51, 4052-4058 (1991).Vitetta E S, Stone M, Amiot P, Fay J, May R, Till M, Newman J, Clark P, Collins R, Cun ningham D, Ghetie V, Uhr J W, Thorpe P E. Phase I immunotoxin trial in patients with B- cell lymphoma. Cancer Res. 51, 4052-4058 (1991).
Wang S-M, Chern J-W, Yeh M-Y, Ng J C, Tung E, Roffler S R. Specific activation of glucuronide prodrugs by antibody-targeted enzyme conjugates for cancer therapy. Cancer Res. 52, 4484-4491 (1992).Wang S-M, Chern J-W, Yeh M-Y, Ng J C, Tung E, Roffler S R. Specific activation of glucuronide prodrugs by antibody-targeted enzyme conjugates for cancer therapy. Cancer Res. 52, 4484-4491 (1992).
Weiner L M, O′Dwyer J, Kitson J, Comis R L, Frankel A E, Bauer R J, Kopnrad M S, Groves E S. Phase I evaluation of an anti-breast carcinoma monoclonal antibody 260F9-re combinant ricin A chain immunoconjugate. Cancer Res. 49, 4062-4067 (1989).Weiner L M, O′Dwyer J, Kitson J, Comis R L, Frankel A E, Bauer R J, Kopnrad M S, Groves E S. Phase I evaluation of an anti-breast carcinoma monoclonal antibody 260F9-re combinant ricin A chain immunoconjugate. Cancer Res. 49, 4062-4067 (1989).
Weiner L M, Holmes M, Adams G P, LaCreta F, Watts P, Garcia de Palzzo I. A human tumor xenograft model of therapy with a bispecific monoclonal antibody targeting c-erbB-2 and CD16. Cancer Res. 53, 94-100 (1993).Weiner L M, Holmes M, Adams G P, LaCreta F, Watts P, Garcia de Palzzo I. A human tumor xenograft model of therapy with a bispecific monoclonal antibody targeting c-erbB-2 and CD16. Cancer Res. 53, 94-100 (1993).
Wunderlich D, Lee A, Fracasso R P, Mierz D V, Bayney R M, Ramabhadran T V. Use of recombinant fusion proteins for generation and rapid characterization of monoclonal anti bodies. J. Immunol Methods 147, 1-11 (1992).Wunderlich D, Lee A, Fracasso R P, Mierz D V, Bayney R M, Ramabhadran T V. Use of recombinant fusion proteins for generation and rapid characterization of monoclonal anti bodies. J. Immunol Methods 147, 1-11 (1992).
Claims (30)
DVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTST
oder eine allele Variante oder ein Fragment dieser Sequenz kodiert.3. Use of an antibody according to claim 2, characterized in that the exon v5 for the amino acid sequence
DVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTST
or encodes an allelic variant or a fragment of this sequence.
GATGTAGACAGAAATGGCACCACTGCTTATGAAGGAAACTGGAACCCAGAAGCACAC CCTCCCCTCATTCACCATGAGCATCATGAGGAAGAAGAGACCCCACATTCTACAAGC ACAA
oder eine degenerierte oder allele Variante oder ein Fragment dieser Sequenz enthält.4. Use of an antibody according to claim 3, characterized in that the exon v5 is the nucleic acid sequence
GATGTAGACAGAAATGGCACCACTGCTTATGAAGGAAACTGGAACCCAGAAGCACAC CCTCCCCTCATTCACCATGAGCATCATGAGGAAGAAGAGACCCCACATTCTACAAGC ACAA
or contains a degenerate or allelic variant or a fragment of this sequence.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4326573A DE4326573A1 (en) | 1993-08-07 | 1993-08-07 | Polypeptides encoded by exon v5 of the CD44 gene as targets for immunotherapy and immunoscintigraphy of tumors |
| AU74598/94A AU7459894A (en) | 1993-08-07 | 1994-07-21 | Polypeptides coded by exon v5 of the cd44 gene as targets for immunotherapy and immunoscintigraphy of tumours |
| PCT/EP1994/002398 WO1995004547A1 (en) | 1993-08-07 | 1994-07-21 | POLYPEPTIDES CODED BY EXON v5 OF THE CD44 GENE AS TARGETS FOR IMMUNOTHERAPY AND IMMUNOSCINTIGRAPHY OF TUMOURS |
| EP94924268A EP0713398A1 (en) | 1993-08-07 | 1994-07-21 | POLYPEPTIDES CODED BY EXON v5 OF THE CD44 GENE AS TARGETS FOR IMMUNOTHERAPY AND IMMUNOSCINTIGRAPHY OF TUMOURS |
| CA002168988A CA2168988A1 (en) | 1993-08-07 | 1994-07-21 | Polypeptides coded by exon v5 of the cd44 gene as targets for immunotherapy and immunoscintigraphy of tumours |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4326573A DE4326573A1 (en) | 1993-08-07 | 1993-08-07 | Polypeptides encoded by exon v5 of the CD44 gene as targets for immunotherapy and immunoscintigraphy of tumors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE4326573A1 true DE4326573A1 (en) | 1995-02-23 |
Family
ID=6494689
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE4326573A Withdrawn DE4326573A1 (en) | 1993-08-07 | 1993-08-07 | Polypeptides encoded by exon v5 of the CD44 gene as targets for immunotherapy and immunoscintigraphy of tumors |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0713398A1 (en) |
| AU (1) | AU7459894A (en) |
| CA (1) | CA2168988A1 (en) |
| DE (1) | DE4326573A1 (en) |
| WO (1) | WO1995004547A1 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997016557A1 (en) * | 1995-10-31 | 1997-05-09 | Boehringer Ingelheim International Gmbh | TREATMENT OF TUMOURS BY ADOPTIVE TRANSFER OF CD44v-SPECIFIC CYTOTOXIC T-LYMPHOCYTES |
| WO1997021104A1 (en) * | 1995-12-06 | 1997-06-12 | Boehringer Ingelheim International Gmbh | Method of diagnosing and treating epithelioma |
| WO1998022508A2 (en) * | 1996-11-21 | 1998-05-28 | Boehringer Ingelheim International Gmbh | Process for tumour cell depletion of cd34-positive cells |
| DE19708713A1 (en) * | 1997-03-04 | 1998-09-17 | Boehringer Ingelheim Int | Use of preparations containing anti-CD44 antibodies for the treatment of certain tumors and for the suppression of immune reactions |
| DE19911329A1 (en) * | 1998-03-27 | 2000-09-21 | Benes Ivan Friedrich | Radioimmunoconjugate which can be used in human therapy and process for its preparation |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7534605B2 (en) | 1999-06-08 | 2009-05-19 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | CD44 polypeptides, polynucleotides encoding same, antibodies directed thereagainst and method of using same for diagnosing and treating inflammatory diseases |
| EP1401472A4 (en) * | 2001-05-25 | 2005-04-06 | Univ Jefferson | Alternative splice forms of proteins as basis for multiple therapeutic modalities |
| PT2886126T (en) | 2013-12-23 | 2017-09-13 | Exchange Imaging Tech Gmbh | Nanoparticle conjugated to cd44 binding peptides |
| BR112017000710B1 (en) | 2014-07-15 | 2024-02-27 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | ISOLATED POLYPEPTIDE, COMPOSITION OF MATTER, AND, USE OF THE ISOLATED POLYPEPTIDE OR COMPOSITION OF MATTER |
| CN116496398B (en) * | 2022-10-31 | 2023-10-31 | 南京元迈细胞生物科技有限公司 | Antibody specifically binding to v5 exon of CD44 and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4014510A1 (en) * | 1990-05-07 | 1991-11-14 | Kernforschungsz Karlsruhe | VARIANT CD44 SURFACE PROTEINS, THESE ENCODING C-DNA SEQUENCES, ANTIBODIES AGAINST THESE PROTEINS AND THEIR USE IN DIAGNOSTICS AND THERAPY |
| DE4134982A1 (en) * | 1991-10-23 | 1993-04-29 | Kernforschungsz Karlsruhe | USE OF ANTIBODY-CONTAINING PREPARATIONS FOR IMMUNE SUPPRESSION |
-
1993
- 1993-08-07 DE DE4326573A patent/DE4326573A1/en not_active Withdrawn
-
1994
- 1994-07-21 CA CA002168988A patent/CA2168988A1/en not_active Abandoned
- 1994-07-21 AU AU74598/94A patent/AU7459894A/en not_active Abandoned
- 1994-07-21 WO PCT/EP1994/002398 patent/WO1995004547A1/en not_active Application Discontinuation
- 1994-07-21 EP EP94924268A patent/EP0713398A1/en not_active Ceased
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997016557A1 (en) * | 1995-10-31 | 1997-05-09 | Boehringer Ingelheim International Gmbh | TREATMENT OF TUMOURS BY ADOPTIVE TRANSFER OF CD44v-SPECIFIC CYTOTOXIC T-LYMPHOCYTES |
| WO1997021104A1 (en) * | 1995-12-06 | 1997-06-12 | Boehringer Ingelheim International Gmbh | Method of diagnosing and treating epithelioma |
| WO1998022508A2 (en) * | 1996-11-21 | 1998-05-28 | Boehringer Ingelheim International Gmbh | Process for tumour cell depletion of cd34-positive cells |
| WO1998022508A3 (en) * | 1996-11-21 | 1998-07-30 | Rupert Handgretinger | Process for tumour cell depletion of cd34-positive cells |
| DE19708713A1 (en) * | 1997-03-04 | 1998-09-17 | Boehringer Ingelheim Int | Use of preparations containing anti-CD44 antibodies for the treatment of certain tumors and for the suppression of immune reactions |
| DE19708713C2 (en) * | 1997-03-04 | 2002-11-28 | Boehringer Ingelheim Int | Use of preparations containing anti-CD44 antibodies for the treatment of certain tumors and for the suppression of immune reactions |
| DE19911329A1 (en) * | 1998-03-27 | 2000-09-21 | Benes Ivan Friedrich | Radioimmunoconjugate which can be used in human therapy and process for its preparation |
Also Published As
| Publication number | Publication date |
|---|---|
| EP0713398A1 (en) | 1996-05-29 |
| CA2168988A1 (en) | 1995-02-16 |
| AU7459894A (en) | 1995-02-28 |
| WO1995004547A1 (en) | 1995-02-16 |
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