Dynamic redistribution of the activating 2B4/SAP complex at the cytotoxic NK cell immune synapse
Dynamic redistribution of the activating 2B4/SAP complex at the cytotoxic NK cell immune synapse
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DOI：
10.4049/jimmunol.173.6.3640
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发表时间：
2004-09-15
影响因子：
4.4
通讯作者：
Fernández-Ruiz, E
中科院分区：
文献类型：
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作者：
Roda-Navarro, P;Mittelbrunn, M;Fernández-Ruiz, E
2B 4分子（CD 244）已被描述为人NK细胞活化中的辅助受体。然而，2134在细胞毒性NK细胞免疫突触（NK-IS）形成过程中的行为仍不确定。在这项研究中，我们证明了在静息NK细胞和EBV感染的721.221人细胞之间形成缀合物后，2B 4和信号转导衔接分子（信号转导淋巴细胞活化分子相关蛋白（SAP））向细胞毒性NK-IS的再分布。共聚焦显微镜显示2B 4定位于中心超分子活化簇，周围为NK细胞内含有talin的外周超分子活化簇和靶细胞上含有ICAM-1的外周超分子活化簇。使用2B 4-GFP转染的NK细胞的视频显微镜研究显示，一旦细胞接触发生，2B 4就会重新分布至细胞毒性NK-IS。同时，SAP-GFP也聚集在接触位点，并在相互作用期间保留在那里。即使在NK细胞脱离后，2B 4分子簇仍与靶细胞结合。这些结果强调了2B 4作为粘附分子的功能，并表明在初始结合、靶细胞扫描和细胞毒性NK-1 S形成中的相关作用。最后，这些发现表明活化2B 4/信号淋巴细胞活化分子相关蛋白复合物在识别EBV感染细胞过程中的重要作用。
The 2B4 molecule (CD244) has been described as a coreceptor in human NK cell activation. However, the behavior of 2134 during the cytotoxic NK cell immune synapse (NK-IS) formation remains undetermined. In this study, we demonstrate the redistribution of 2B4 and the signaling adaptor molecule, signaling lymphocyte activation molecule-associated protein (SAP), to the cytotoxic NK-IS upon formation of conjugates between resting NK cells and EBV-infected 721.221 human cells. Confocal microscopy showed that 2B4 localized at the central supramolecular activation cluster, surrounded by a peripheral supramolecular activation cluster containing talin within NK cell and ICAM-1 on target cells. Videomicroscopy studies with 2B4-GFP-transfected NK cells revealed that 2B4 redistributed to cytotoxic NK-IS as soon as the cell contact occurred. Simultaneously, a SAP-GFP also clustered at the contact site, where it remained during the interaction period. The 2B4 molecular clusters remained bound to the target cell even after NK cell detachment. These results underscore the function of 2B4 as an adhesion molecule and suggest a relevant role in the initial binding, scanning of target cells, and formation of cytotoxic NK-IS. Finally, these findings are indicative of an important role of the activating 2B4/signaling lymphocyte activation molecule-associated protein complex during the recognition of EBV-infected cells.