An anti-alpha v-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice.
A series of murine monoclonal antibodies were raised against purified human alpha v beta 3 integrin and against M21 human melanoma cells. Five notable hybridomas were identified by ELISA on purified integrins, and the isolated antibodies bound the alpha v-chain. These antibodies, 17E6, 20A9, 23G5, 14D9.F8 and 10G2, recognised the extracellular domains of the integrin, and were shown to be reactive in FACS, immunoprecipitation, ELISA, and ELISA on fixed cells with M21, M21-L4, and UCLA-P3, but not with the alpha v-deficient M21-L or M21-L-IIb (M21-L transfected with GpIIb integrin). One antibody, 17E6, strongly perturbed cell attachment mediated by alpha v integrins, reacting at least with alpha v beta 3, alpha v beta 5, and alpha v beta 1, and strongly inhibiting cell attachment to alpha v-ligands vitronectin and fibronectin with an IC50 of approximately 0.1 microgram ml-1. Furthermore, 17E6 at this concentration could induce cell retraction from the substrate, while LM609 (anti-alpha v beta 3) and control antibody 14E2 (anti-200 kDa melanoma surface protein) at 1,000-fold higher concentrations had minimal effects on cell morphology. The action of 17E6 was reversible and was not due to toxic effects: in vitro 17E6 at 0.1 mg ml-1 did not affect either cell proliferation or DNA synthesis. In two nude-mouse tumour models, subcutaneous tumour development and a lung colonisation ('experimental metastasis') assay, injection of 17E6 strongly inhibited tumour development, while isotype-matched controls had no effect. There was no obvious mechanism of cell or of complement-mediated tumour cytotoxicity; the antibody did not mediate ADCC or AECDC, or complement fixation. The data strongly support previous studies which have indicated the importance of alpha v-integrins, and especially alpha v beta 3, in the tumour progression of human melanoma.
Mitjans
F
F
Merck Farma y Quimica, Laboratorio de Bioinvestigaciòn (LBI), Barcelona, Spain.
Sander
D
D
Adán
J
J
Sutter
A
A
Martinez
J M
JM
Jäggle
C S
CS
Moyano
J M
JM
Kreysch
H G
HG
Piulats
J
J
Goodman
S L
SL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000920:Antibody-Dependent Cell Cytotoxicity; D000936:Antigen-Antibody Complex; D002460:Cell Line; D004261:DNA Replication / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D007145:Immunoglobulin kappa-Chains / Q000494:pharmacology; D008545:Melanoma / Q000276:immunology / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008819:Mice, Nude; D011233:Precipitin Tests; D019030:Receptors, Vitronectin / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D019030:Receptors, Vitronectin