Kiener P A, Mittler R S
Bristol-Myers Company, Department of Immunology, Wallingford, CT 06492-7660.
J Immunol. 1989 Jul 1;143(1):23-8.
Activation of peripheral blood T cells, and the leukemic T cell line Jurkat, as measured by mobilization of intracellular calcium, by an anti-TCR antibody is blocked by mAb (T191) to the leukocyte common Ag (CD45). T191 also blocked down-regulation of the CD3-TCR complex induced by an anti-CD3 mAb. Vanadate, a phosphotyrosine phosphatase inhibitor, partially blocks the effect of T191 and restored mobilization of intracellular calcium. Assays of the immunoprecipitates of T191 and CD45 from both Jurkat-BM1 and peripheral T cells showed that the immune complexes had intrinsic phosphatase activity. A parallel immunoprecipitate using a mAb (4-10) against HLA class I showed no such activity. Further analysis of the T191 immunocomplex revealed activity against phosphotyrosine, p-nitrophenylphosphate, and [32P-poly-glu-tyr, but not against phosphoserine. Phosphatase activity was inhibited by Vanadate, but not by Zn2+ or F-. These results show that CD45 is a phosphotyrosine phosphatase, and strongly suggest that tyrosine phosphorylation/dephosphorylation is critically involved in activation of T cells through the TCR-CD3 complex.
通过细胞内钙动员来检测,抗TCR抗体对外周血T细胞和白血病T细胞系Jurkat的激活作用,被针对白细胞共同抗原(CD45)的单克隆抗体(T191)所阻断。T191也阻断了抗CD3单克隆抗体诱导的CD3 - TCR复合物的下调。钒酸盐是一种磷酸酪氨酸磷酸酶抑制剂,它部分阻断了T191的作用并恢复了细胞内钙的动员。对来自Jurkat - BM1和外周T细胞的T191和CD45免疫沉淀物的检测表明,免疫复合物具有内在的磷酸酶活性。使用针对HLA I类的单克隆抗体(4 - 10)进行的平行免疫沉淀未显示出这种活性。对T191免疫复合物的进一步分析揭示了其对磷酸酪氨酸、对硝基苯磷酸酯和[32P - 聚 - 谷 - 酪]的活性,但对磷酸丝氨酸无活性。磷酸酶活性被钒酸盐抑制,但不被Zn2+或F - 抑制。这些结果表明CD45是一种磷酸酪氨酸磷酸酶,并强烈提示酪氨酸磷酸化/去磷酸化在通过TCR - CD3复合物激活T细胞过程中起关键作用。