ICAM-3 REGULATES LYMPHOCYTE MORPHOLOGY AND INTEGRIN-MEDIATED T-CELL INTERACTION WITH ENDOTHELIAL-CELL AND EXTRACELLULAR-MATRIX LIGANDS
ICAM-3 REGULATES LYMPHOCYTE MORPHOLOGY AND INTEGRIN-MEDIATED T-CELL INTERACTION WITH ENDOTHELIAL-CELL AND EXTRACELLULAR-MATRIX LIGANDS
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DOI：
10.1083/jcb.127.3.867
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发表时间：
1994-11-01
影响因子：
7.8
通讯作者：
SANCHEZMADRID, F
中科院分区：
文献类型：
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作者：
CAMPANERO, MR;SANCHEZMATEOS, P;SANCHEZMADRID, F
白细胞活化是一个复杂的过程，涉及多个交叉调节的细胞粘附事件。在这份报告中，我们研究了细胞间粘附分子-3（ICAM-3），β 2整合素白细胞功能相关抗原-1（LFA-1）的第三种已鉴定配体，在调节白细胞粘附ICAM-1、血管细胞粘附分子-1（VCAM-1）和纤连蛋白38-和80-kD片段（FN 40和FN 80）中的作用。当与非常低剂量的抗CD 3 mAb组合时，活化性抗ICAM-3 HP 2/19而不是其它抗ICAM-3 mAb能够增强T淋巴母细胞与这些蛋白的粘附，所述抗CD 3 mAb本身不能诱导这种现象。与此相反，抗ICAM-1单克隆抗体并没有增强T细胞附着到这些基质。抗LFA-1、抗VLA α 4和抗VLA α 5 mAb分别特异性阻断T细胞与ICAM-1、VCAM-1、FN 40和FN 80的粘附。活化性抗ICAM-3 HP 2/19还能够特异性增强VLA-4和VLA-5介导的白血病T Jurkat细胞与VCAM-1、FN 40和FN 80的结合，即使在不存在CD 3-TcR复合物共占有的情况下。我们还通过免疫荧光显微镜研究了ICAM-3、LFA-1和VLA β 1整合素在与ICAM-1、VCAM-1和FN 80相互作用的细胞上的定位。我们发现，抗ICAM-3 HP 2/19单克隆抗体特异性地促进了T淋巴母细胞的形态发生显著变化时，这些细胞被允许与这些粘附配体相互作用。在这些条件下，观察到一个大的细胞接触面积，从该面积向外环境投射一个尾足类结构（头尾足类）。然而，当用其他粘附促进剂如活化性抗VLA β 1 TS 2/16 mAb或佛波醇酯刺激T母细胞时，未检测到这种结构。抗ICAM-3 TP 1/24 mAb也不能诱导这种现象。值得注意的是，HP 2/19 mAb特异性地诱导ICAM-3的显著细胞再分布，但其他抗ICAM-3 mAb或其他促粘附剂不诱导。因此，ICAM-3几乎完全集中在最远端部分的头尾足，而无论是LFA-1或VLA β 1整联蛋白均匀分布在整个大的接触面积。此外，当用HP 2/19 mAb特异性刺激T细胞与TNF α激活的内皮细胞相互作用时，也观察到这种现象。我们发现了肌球蛋白的线性阵列内的标题尾足动物的本地化。相反，肌动蛋白为基础的细胞骨架呈现出均匀的分布在广泛的接触面积与基板。此外，丁二酮单肟，肌球蛋白破坏药物，取消了形态学细胞的变化和ICAM-3聚集。总之，这些结果表明ICAM-3对T细胞粘附和形态的多个途径具有调节作用。
Leukocyte activation is a complex process that involves multiple cross-regulated cell adhesion events. In this report, we investigated the role of intercellular adhesion molecule-3 (ICAM-3), the third identified ligand for the beta 2 integrin leukocyte function-associated antigen-1 (LFA-1), in the regulation of leukocyte adhesion to ICAM-1, vascular cell adhesion molecule-1 (VCAM-1), and the 38- and 80-kD fragments of fibronectin (FN40 and FN80). The activating anti-ICAM-3 HP2/19, but not other anti-ICAM-3 mAb, was able to enhance T lymphoblast adhesion to these proteins when combined with very low doses of anti-CD3 mAb, which were unable by themselves to induce this phenomenon. In contrast, anti-ICAM-1 mAb did not enhance T cell attachment to these substrata. T cell adhesion to ICAM-1, VCAM-1, FN40, and FN80 was specifically blocked by anti-LFA-1, anti-VLA alpha 4, and anti-VLA alpha 5 mAb, respectively. The activating anti-ICAM-3 HP2/19 was also able to specifically enhance the VLA-4- and VLA-5-mediated binding of leukemic T Jurkat cells to VCAM-1, FN40, and FN80, even in the absence of cooccupancy of the CD3-TcR complex. We also studied the localization of ICAM-3, LFA-1, and the VLA beta 1 integrin, by immunofluorescence microscopy, on cells interacting with ICAM-1, VCAM-1 and FN80. We found that the anti-ICAM-3 HP2/19 mAb specifically promoted a dramatic change on the morphology of T lymphoblasts when these cells were allowed to interact with those adhesion ligands. Under these conditions, it was observed that a large cell contact area from which an uropod-like structure (heading uropod) was projected toward the outer milieu. However, when T blasts were stimulated with other adhesion promoting agents as the activating anti-VLA beta 1 TS2/16 mAb or phorbol esters, this structure was not detected. The anti-ICAM-3 TP1/24 mAb was also unable to induce this phenomenon. Notably, a striking cell redistribution of ICAM-3 was induced specifically by the HP2/19 mAb, but not by the other anti-ICAM-3 mAb or the other adhesion promoting agents. Thus, ICAM-3 was almost exclusively concentrated in the most distal portion of the heading uropod whereas either LFA-1 or the VLA beta 1 integrin were uniformly distributed all over the large contact area. Moreover, this phenomenon was also observed when T cells were specifically stimulated with the HP2/19 mAb to interact with TNF alpha-activated endothelial cells. We found the localization of linear arrays of myosin within the heading uropod. In contrast, actin-based cytoskeleton presented a uniform distribution over the broad contact area with the substrate. In addition, butanedione monoxime, a myosin-disrupting drug, abolished both the morphological cell change and ICAM-3 clustering. Altogether, these results demonstrate that ICAM-3 has a regulatory role on multiple pathways of T cell adhesion and morphology.