del Pozo M A, Cabañas C, Montoya M C, Ager A, Sánchez-Mateos P, Sánchez-Madrid F
Servicio de Immunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Spain.
J Cell Biol. 1997 Apr 21;137(2):493-508. doi: 10.1083/jcb.137.2.493.
The recruitment of leukocytes from the bloodstream is a key step in the inflammatory reaction, and chemokines are among the main regulators of this process. During lymphocyte-endothelial interaction, chemokines induce the polarization of T lymphocytes, with the formation of a cytoplasmic projection (uropod) and redistribution of several adhesion molecules (ICAM-1,-3, CD43, CD44) to this structure. Although it has been reported that these cytokines regulate the adhesive state of integrins in leukocytes, their precise mechanisms of chemoattraction remain to be elucidated. We have herein studied the functional role of the lymphocyte uropod. Confocal microscopy studies clearly showed that cell uropods project away from the cell bodies of adhered lymphocytes and that polarized T cells contact other T cells through the uropod structure. Time-lapse videomicroscopy studies revealed that uropod-bearing T cells were able, through this cellular projection, to contact, capture, and transport additional bystander T cells. Quantitative analysis revealed that the induction of uropods results in a 5-10-fold increase in cell recruitment. Uropod-mediated cell recruitment seems to have physiological relevance, since it was promoted by both CD45R0+ peripheral blood memory T cells as well as by in vivo activated lymphocytes. Additional studies showed that the cell recruitment mediated by uropods was abrogated with antibodies to ICAM-1, -3, and LFA-1, whereas mAb to CD43, CD44, CD45, and L-selectin did not have a significant effect, thus indicating that the interaction of LFA-1 with ICAM-1 and -3 appears to be responsible for this process. To determine whether the increment in cell recruitment mediated by uropod may affect the transendothelial migration of T cells, we carried out chemotaxis assays through confluent monolayers of endothelial cells specialized in lymphocyte extravasation. An enhancement of T cell migration was observed under conditions of uropod formation, and this increase was prevented by incubation with either blocking anti-ICAM-3 mAbs or drugs that impair uropod formation. These data indicate that the cell interactions mediated by cell uropods represent a cooperative mechanism in lymphocyte recruitment, which may act as an amplification system in the inflammatory response.
白细胞从血流中募集是炎症反应的关键步骤,趋化因子是这一过程的主要调节因子之一。在淋巴细胞与内皮细胞相互作用期间,趋化因子诱导T淋巴细胞极化,形成细胞质突起(尾足),并使几种黏附分子(ICAM-1、-3、CD43、CD44)重新分布到该结构上。尽管已有报道称这些细胞因子调节白细胞中整合素的黏附状态,但其精确的趋化机制仍有待阐明。我们在此研究了淋巴细胞尾足的功能作用。共聚焦显微镜研究清楚地表明,细胞尾足从黏附淋巴细胞的细胞体伸出,极化的T细胞通过尾足结构与其他T细胞接触。延时视频显微镜研究显示,带有尾足的T细胞能够通过这种细胞突起接触、捕获并转运其他旁观T细胞。定量分析表明,尾足的诱导导致细胞募集增加5至10倍。尾足介导的细胞募集似乎具有生理相关性,因为CD45R0 +外周血记忆T细胞以及体内活化的淋巴细胞均可促进其发生。进一步的研究表明,尾足介导的细胞募集可被抗ICAM-1、-3和LFA-1抗体消除,而抗CD43、CD44、CD45和L-选择素的单克隆抗体则无显著作用,这表明LFA-1与ICAM-1和-3的相互作用似乎是这一过程的原因。为了确定尾足介导的细胞募集增加是否会影响T细胞的跨内皮迁移,我们通过专门用于淋巴细胞外渗的内皮细胞汇合单层进行了趋化性测定。在尾足形成的条件下观察到T细胞迁移增强,并且通过与阻断性抗ICAM-3单克隆抗体或损害尾足形成的药物孵育可阻止这种增加。这些数据表明,细胞尾足介导的细胞相互作用代表了淋巴细胞募集中的一种协同机制,可能在炎症反应中作为一种放大系统发挥作用。