Finkelstein Lisa D, Shimizu Yoji, Schwartzberg Pamela L
National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
J Immunol. 2005 Nov 1;175(9):5923-30. doi: 10.4049/jimmunol.175.9.5923.
T cells deficient in the Tec kinases Itk or Itk and Rlk exhibit defective TCR-stimulated proliferation, IL-2 production, and activation of phospholipase C-gamma. Evidence also implicates Tec kinases in actin cytoskeleton regulation, which is necessary for cell adhesion and formation of the immune synapse in T lymphocytes. In this study we show that Tec kinases are required for TCR-mediated up-regulation of adhesion via the LFA-1 integrin. We also demonstrate that the defect in adhesion is associated with defective clustering of LFA-1 and talin at the site of interaction of Rlk-/-Itk-/- and Itk-/- T cells with anti-TCR-coated beads. Defective recruitment of Vav1, protein kinase Ctheta, and Pyk2 was also observed in Rlk-/-Itk-/- and Itk-/- T cells. Stimulation with ICAM-2 in conjunction with anti-TCR-coated beads enhanced polarization of Vav1, protein kinase Ctheta, and Pyk2 in wild-type cells, demonstrating a role for integrins in potentiating the recruitment of signaling molecules in T cells. Increased recruitment of signaling molecules was most pronounced under conditions of low TCR stimulation. Under these suboptimal TCR stimulation conditions, ICAM-2 could also enhance the recruitment of signaling molecules in Itk-/-, but not Rlk-/-Itk-/- T cells. Thus, Tec kinases play key roles in regulating TCR-mediated polarization of integrins and signaling molecules to the site of TCR stimulation as well as the up-regulation of integrin adhesion.
缺乏Tec激酶Itk或同时缺乏Itk和Rlk的T细胞在TCR刺激下表现出增殖缺陷、白细胞介素-2产生缺陷以及磷脂酶C-γ激活缺陷。有证据表明Tec激酶也参与肌动蛋白细胞骨架调节,这对于T淋巴细胞中的细胞黏附和免疫突触形成是必需的。在本研究中,我们表明Tec激酶是TCR介导的通过淋巴细胞功能相关抗原-1(LFA-1)整合素上调黏附所必需的。我们还证明,Rlk基因敲除/Itk基因敲除和Itk基因敲除的T细胞与抗TCR包被珠子相互作用部位的LFA-1和踝蛋白聚集缺陷与黏附缺陷相关。在Rlk基因敲除/Itk基因敲除和Itk基因敲除的T细胞中也观察到Vav1、蛋白激酶Cθ和黏着斑激酶2(Pyk2)募集缺陷。用细胞间黏附分子-2(ICAM-2)联合抗TCR包被珠子刺激可增强野生型细胞中Vav1、蛋白激酶Cθ和Pyk2的极化,表明整合素在增强T细胞中信号分子募集中的作用。信号分子募集增加在低TCR刺激条件下最为明显。在这些次优TCR刺激条件下,ICAM-2也可增强Itk基因敲除T细胞中信号分子的募集,但不能增强Rlk基因敲除/Itk基因敲除T细胞中信号分子的募集。因此,Tec激酶在调节TCR介导的整合素和信号分子向TCR刺激部位的极化以及整合素黏附上调中起关键作用。