Cytotoxic T lymphocytes express a beta(3) integrin which can induce the phosphorylation of focal adhesion kinase and the related PYK-2
Cytotoxic T lymphocytes express a beta(3) integrin which can induce the phosphorylation of focal adhesion kinase and the related PYK-2
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DOI：
10.1002/eji.1830270147
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发表时间：
1997-01-01
影响因子：
5.4
通讯作者：
Ostergaard, HL
中科院分区：
文献类型：
--
作者：
Ma, EA;Lou, O;Ostergaard, HL
纤维连接蛋白已被证明可以刺激115-125 kDa范围内的许多蛋白质的酪氨酸磷酸化，并促进同种抗原特异性细胞毒性T淋巴细胞（CTL）克隆对亚刺激量的抗cd3或抗T细胞受体（TCR）的反应。目前的研究是为了进一步表征这些CTL克隆的整合素表达和使用。我们证明，玻璃体粘连蛋白和纤维蛋白原，而不是层粘连蛋白或胶原蛋白，也能够在亚刺激抗cd3存在的情况下促进脱粒，并刺激这些115-125 kda蛋白的酪氨酸磷酸化，其中115-kDa蛋白的磷酸化最为显著。这些结果暗示这些CTL克隆表达和使用玻璃体连接蛋白受体α - β 3整合素。我们通过流式细胞术和免疫沉淀证明CTL克隆确实表达β(3)整合素。固定的β(3)抗体刺激115-125-kDa蛋白的磷酸化，表明β(3)与纤维连接蛋白或玻璃体连接蛋白的结合将相同的信号传递到这些细胞中。纤维连接蛋白和玻璃体连接蛋白诱导的磷酸化以及与纤维连接蛋白或玻璃体连接蛋白的粘附可以被β(3)整合素抗体显著抑制。最后，我们能够用抗血清将115-kDa蛋白免疫沉淀到局灶黏附激酶和一种相关的激酶PYK-2， PYK-2在玻璃化粘连蛋白或固定化抗β蛋白的作用下被磷酸化(3)。综上所述，这些结果表明CTL表达并使用β(3)-整合素作为信号分子，可以增强tcr介导的刺激。
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 beta 3 integrin, by these CTL clones. We demonstrate by both flow cytometry and immunoprecipitation that CTL clones do in fact express beta(3) integrin. Immobilized antibody to beta(3) stimulates the phosphorylation of the 115-125-kDa proteins, suggesting that engagement of beta(3) 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 beta(3) 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-beta(3). Taken together, these results demonstrate that CTL express and use beta(3)-integrins as signaling molecules which can augment TCR-mediated stimulation.