The Role of Co-receptors in T-cell Actin Barrier Evasion during HIV Entry

描述(申请人提供)：最近的研究表明，人类免疫缺陷病毒(HIV)能够感染宿主中大量存在并通常对病毒具有相当抵抗力的静息T细胞。这种抵抗力至少部分是由T细胞质膜附近一层厚厚的基于肌动蛋白的细胞骨架提供的，这通常会阻止病毒颗粒与宿主细胞完全融合，并将病毒基因组传递到细胞内。然而，病毒偶尔能够通过启动来绕过这一防御 激活细胞自身的肌动蛋白调节酶以削弱细胞骨架层的细胞信号级联。该项目的目标是(1)准确地找出HIV使用这些酶中的哪一种来削弱T细胞的肌动蛋白层，以及(2)确定每种酶被招募到HIV进入部位的顺序。这些知识是使未来的工作能够找到一种方法来加强T细胞作为一种新型的艾滋病毒/艾滋病抑制物自身防御所需的基础。为了获得这一信息，来自实验室的T细胞系和原代T细胞将用超分辨率荧光显微镜成像。已知或可能参与HIV用来拆除肌动蛋白进入屏障的信号过程的各种细胞蛋白质将被标记上荧光标签。这些标记蛋白的定位将在活细胞中观察，并使用先进的基于单分子的颗粒跟踪和超分辨率成像技术以高分辨率确定。

DESCRIPTION (provided by applicant): Recent research has shown that the human immunodeficiency virus (HIV) is able to infect resting T cells, which are present in large number in the host and generally quite resistant to the virus. This resistance is at least partially provied by a thick layer of actin-based cytoskeleton near the T cell's plasma membrane that usually prevents the viral particles from fully fusing with the host cell and delivering the viral genome into the cell. However, viruses are able to occasionally circumvent this defense through initiation of a cellular signaling cascade that activates the cell's own actin-regulating enzymes to weaken the cytoskeleton layer. The goal of this project is to (1) find exactly which of these enzymes HIV uses to weaken the T cell's actin layer and (2) determine the order that each enzyme is recruited to the site of HIV entry. This knowledge is the foundation that is needed to enable future work on finding a way to strengthen the T cells' own defense as a new type of HIV/AIDS inhibition. To obtain this information, T-cell derived laboratory cell lines and primary T cells wil be imaged with super-resolution fluorescence microscopy. Various cellular proteins that are known or proposed to be involved with the signaling process that HIV uses to dismantle the actin barrier to entry will be labeled with fluorescent tags. The localization of these tagged proteins will be watched in live cells and determined with high resolution using advanced single- molecule based particle tracking and super-resolution imaging techniques.