HIV-1 Nef Inhibits Ruffles, Induces Filopodia, and Modulates Migration of Infected Lymphocytes
HIV-1 Nef Inhibits Ruffles, Induces Filopodia, and Modulates Migration of Infected Lymphocytes
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DOI：
10.1128/jvi.02230-09
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发表时间：
2010-03-01
影响因子：
5.4
通讯作者：
Sol-Foulon, Nathalie
中科院分区：
文献类型：
--
作者：
Nobile, Cinzia;Rudnicka, Dominika;Sol-Foulon, Nathalie
HIV-1 Nef 蛋白是调节受感染细胞行为的致病因子。 Nef 诱导肌动蛋白细胞骨架变化并损害细胞向趋化因子的迁移。我们进一步表征了 HIV-1 感染淋巴细胞的形态、细胞骨架动力学和运动性。通过使用扫描电子显微镜、共焦免疫荧光显微镜和结合流式细胞术和自动成像的 ImageStream 技术，我们报告 HIV-1 诱导肌动蛋白细胞骨架的特征性重塑。在受感染的淋巴细胞中，褶皱的形成受到抑制，而长而细的丝状足样突起被诱导。感染 HIV 且删除了 nef 的细胞表现出正常的表型，并且在没有其他病毒蛋白的情况下，单独表达 Nef 会诱导形态变化。我们还使用创新的成像系统来固定和可视化悬浮液中的活个体细胞。当与共焦“轴向断层扫描”相结合时，该技术极大地提高了三维光学分辨率。通过这项技术，我们证实了在未固定的表达 Nef 的淋巴细胞中诱导了长丝状足结构。 Nef 诱导的细胞骨架重组与细胞运动的重要损伤有关。感染细胞对纤连蛋白的粘附和扩散、自发运动以及向趋化因子（CXCL12、CCL3 和 CCL19）的迁移均显着降低。因此，Nef 对淋巴细胞肌动蛋白细胞骨架和细胞形态产生复杂的影响，这可能会影响受感染细胞循环以及与旁观者细胞相遇和交流的能力。
The HIV-1 Nef protein is a pathogenic factor modulating the behavior of infected cells. Nef induces actin cytoskeleton changes and impairs cell migration toward chemokines. We further characterized the morphology, cytoskeleton dynamics, and motility of HIV-1-infected lymphocytes. By using scanning electron microscopy, confocal immunofluorescence microscopy, and ImageStream technology, which combines flow cytometry and automated imaging, we report that HIV-1 induces a characteristic remodeling of the actin cytoskeleton. In infected lymphocytes, ruffle formation is inhibited, whereas long, thin filopodium-like protrusions are induced. Cells infected with HIV with nef deleted display a normal phenotype, and Nef expression alone, in the absence of other viral proteins, induces morphological changes. We also used an innovative imaging system to immobilize and visualize living individual cells in suspension. When combined with confocal "axial tomography," this technique greatly enhances three-dimensional optical resolution. With this technique, we confirmed the induction of long filopodium-like structures in unfixed Nef-expressing lymphocytes. The cytoskeleton reorganization induced by Nef is associated with an important impairment of cell movements. The adhesion and spreading of infected cells to fibronectin, their spontaneous motility, and their migration toward chemokines (CXCL12, CCL3, and CCL19) were all significantly decreased. Therefore, Nef induces complex effects on the lymphocyte actin cytoskeleton and cellular morphology, which likely impacts the capacity of infected cells to circulate and to encounter and communicate with bystander cells.