Identification and functional characterization of mouse CD29 with a mAb.
The beta 1 integrin subfamily, alternatively called very late activation antigen (VLA), has been implicated in various cellular functions. In this study, we generated a mAb against the mouse beta 1 subunit (CD29) to examine the functional property of mouse VLA proteins. After immunization with affinity-purified mouse VLA-4 (alpha 4 beta 1), a hamster mAb, HM beta 1-1, was established by screening mAb that reacted with alpha 4-negative neuroblastoma C1300. The antigen defined by HM beta 1-1 was widely distributed in various mouse cell lines and HM beta 1-1 immunoprecipitated a 110-120 kDa protein common to VLA-1 and VLA-6, indicating that HM beta 1-1 recognizes the beta 1 subunit of mouse integrins. We then examined the inhibitory effect of HM beta 1-1 on VLA-dependent cell adhesion and activation. HM beta 1-1 blocked the adhesion of mouse tumor cell lines to extracellular matrix proteins including collagen, laminin and fibronectin. Moreover, splenic T cell proliferation induced by anti-CD3 mAb and allogeneic mixed lymphocyte response were strongly inhibited by HM beta 1-1 in combination with an anti-LFA-1 mAb. We conclude that HM beta 1-1 reactive with mouse CD29 can inhibit VLA-dependent cellular functions and, thus, would be useful for studying the physiological role of beta 1 integrins in vivo.
Noto
K
K
Department of Respiratory Medicine, Juntendo University School of Medicine, Tokyo.
Kato
K
K
Okumura
K
K
Yagita
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive; D017252:CD3 Complex / Q000276:immunology; D002448:Cell Adhesion / Q000276:immunology; D006224:Cricetinae; D019012:Integrin beta1 / Q000737:chemistry / Q000276:immunology* / Q000502:physiology; D007519:Isoantigens / Q000276:immunology; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D013601:T-Lymphocytes / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D019012:Integrin beta1; D007519:Isoantigens