D'Souza-Schorey C, Boettner B, Van Aelst L
Department of Cell Biology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Mol Cell Biol. 1998 Jul;18(7):3936-46. doi: 10.1128/MCB.18.7.3936.
Leukocyte adhesion to the extracellular matrix (ECM) is tightly controlled and is vital for the immune response. Circulating lymphocytes leave the bloodstream and adhere to ECM components at sites of inflammation and lymphoid tissues. Mechanisms for regulating T-lymphocyte-ECM adhesion include (i) an alteration in the affinity of cell surface integrin receptors for their extracellular ligands and (ii) an alteration of events following postreceptor occupancy (e.g., cell spreading). Whereas H-Ras and R-Ras were previously shown to affect T-cell adhesion by altering the affinity state of the integrin receptors, no signaling molecule has been identified for the second mechanism. In this study, we demonstrated that expression of an activated mutant of Rac triggered dramatic spreading of T cells and their increased adhesion on immobilized fibronectin in an integrin-dependent manner. This effect was not mimicked by expression of activated mutant forms of Rho, Cdc42, H-Ras, or ARF6, indicating the unique role of Rac in this event. The Rac-induced spreading was accompanied by specific cytoskeletal rearrangements. Also, a clustering of integrins at sites of cell adhesion and at the peripheral edges of spread cells was observed. We demonstrate that expression of RacV12 did not alter the level of expression of cell surface integrins or the affinity state of the integrin receptors. Moreover, our results indicate that Rac plays a role in the regulation of T-cell adhesion by a mechanism involving cell spreading, rather than by altering the level of expression or the affinity of the integrin receptors. Furthermore, we show that the Rac-mediated signaling pathway leading to spreading of T lymphocytes did not require activation of c-Jun kinase, serum response factor, or pp70(S6 kinase) but appeared to involve a phospholipid kinase.
白细胞与细胞外基质(ECM)的黏附受到严格调控,对免疫反应至关重要。循环中的淋巴细胞离开血液,在炎症部位和淋巴组织处黏附于ECM成分。调节T淋巴细胞与ECM黏附的机制包括:(i)细胞表面整合素受体对其细胞外配体的亲和力改变;(ii)受体占据后发生的事件改变(如细胞铺展)。尽管先前已表明H-Ras和R-Ras通过改变整合素受体的亲和力状态来影响T细胞黏附,但尚未鉴定出参与第二种机制的信号分子。在本研究中,我们证明激活的Rac突变体的表达以整合素依赖的方式引发T细胞的显著铺展及其在固定化纤连蛋白上黏附增加。Rho、Cdc42、H-Ras或ARF6的激活突变体形式的表达未模拟此效应,表明Rac在此事件中的独特作用。Rac诱导的铺展伴随着特定的细胞骨架重排。此外,在细胞黏附部位和铺展细胞的周边边缘观察到整合素聚集。我们证明RacV12的表达未改变细胞表面整合素的表达水平或整合素受体的亲和力状态。此外,我们的结果表明,Rac通过涉及细胞铺展的机制而非通过改变整合素受体的表达水平或亲和力来调节T细胞黏附。此外,我们表明导致T淋巴细胞铺展的Rac介导的信号通路不需要c-Jun激酶、血清反应因子或pp70(S6激酶)的激活,但似乎涉及一种磷脂激酶。