Cytotoxic T lymphocytes express a beta3 integrin which can induce the phosphorylation of focal adhesion kinase and the related PYK-2.
Fibronectin has been shown to stimulate tyrosine phosphorylation of a number of proteins in the 115-125 kDa range and facilitate degranulation by alloantigen-specific cytotoxic T lymphocyte (CTL) clones in response to substimulatory amounts of anti-CD3 or anti-T cell receptor (TCR). The current study was initiated to further characterize integrin expression and usage by these CTL clones. We demonstrate that vitronectin and fibrinogen, but not laminin or collagen, are also able to both facilitate degranulation in the presence of substimulatory anti-CD3 and stimulate tyrosine phosphorylation of these 115-125-kDa proteins, with a 115-kDa protein being the most prominently phosphorylated. These results implicate the expression and usage of the vitronectin receptor, alpha beta3 integrin, by these CTL clones. We demonstrate by both flow cytometry and immunoprecipitation that CTL clones do in fact express beta3 integrin. Immobilized antibody to beta3 stimulates the phosphorylation of the 115-125-kDa proteins, suggesting that engagement of beta3 transmits the same signal into these cells as fibronectin or vitronectin. The fibronectin and vitronectin-induced phosphorylation as well as adhesion to either fibronectin or vitronectin can be significantly inhibited with antibodies to beta3 integrins. Finally, we are able to immunoprecipitate 115-kDa proteins with antiserum to focal adhesion kinase and a related kinase, called PYK-2, that becomes phosphorylated in response to vitronectin or immobilized anti-beta3. Taken together, these results demonstrate that CTL express and use beta3-integrins as signaling molecules which can augment TCR-mediated stimulation.
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Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
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Ostergaard
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D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D015550:Cell Degranulation; D002999:Clone Cells; D005340:Fibrinogen / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D051417:Focal Adhesion Kinase 1; D051418:Focal Adhesion Kinase 2; D051416:Focal Adhesion Protein-Tyrosine Kinases; D016130:Immunophenotyping; D039661:Integrin beta3; D008213:Lymphocyte Activation; D051379:Mice; D008970:Molecular Weight; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D015398:Signal Transduction; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism*; D019096:Vitronectin / Q000378:metabolism
D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D005353:Fibronectins; D039661:Integrin beta3; D010750:Phosphoproteins; D010980:Platelet Membrane Glycoproteins; D019096:Vitronectin; D019000:Phosphotyrosine; D005340:Fibrinogen; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051418:Focal Adhesion Kinase 2; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; C496053:Ptk2b protein, mouse