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

HIV 进入过程中辅助受体在 T 细胞肌动蛋白屏障逃避中的作用

• 批准号: 9020980
• 负责人: Brian Thomas DeVree
• 金额: $ 5.23万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9020980
• 关键词: Acquired Immunodeficiency Syndrome; Actins; Anti-Retroviral Agents; Binding; CCR5 gene; CD4 Antigens; CXCR4 gene; Cell Line; Cell membrane; Cell surface; Cells; Cellular biology; Chemotaxis; Complex; Cytoskeleton; Development; Enzymes; Fluorescence Microscopy; Foundations; Future; Goals; HIV; Health; Image; Imaging Techniques; Infection; Kinetics; Knowledge; LIMK1 gene; Label; Laboratories; Leukocyte Chemotaxis; Life; Ligands; Mediating; Microscopy; Modeling; Movement; Natural Resistance; Outcome; Pathway interactions; Population; Process; Proteins; Proviruses; Receptor Signaling; Recruitment Activity; Reporting; Research; Resistance; Resistance to infection; Resolution; Rest; Role; Signal Transduction; Signaling Protein; Site; T-Lymphocyte; Techniques; Thick; Time; Total Internal Reflection Fluorescent; Transformed Cell Line; Viral; Viral Genome; Virion; Virus; Virus Diseases; Work; base; cofilin; depolymerization; env Gene Products; image reconstruction; interest; live cell imaging; mortality; new therapeutic target; particle; prevent; protective effect; rac1 GTP-Binding Protein; receptor; receptor-mediated signaling; signal processing; single molecule; therapeutic development; transmission process; virus envelope

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.

描述（由申请人提供）：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的静息T细胞，并且通常对病毒具有相当的抗性。这种抗性至少部分地由T细胞质膜附近的基于肌动蛋白的细胞骨架的厚层提供，所述细胞骨架通常阻止病毒颗粒与宿主细胞完全融合并将病毒基因组递送到细胞中。然而，病毒偶尔能够通过启动来绕过这种防御 这是一种细胞信号级联反应，激活细胞自身的肌动蛋白调节酶，削弱细胞骨架层。该项目的目标是（1）准确地找到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）确定每个酶被招募到HIV进入位点的顺序。这一知识是未来工作所需的基础，以便找到一种方法来加强T细胞自身的防御，作为一种新型的HIV/AIDS抑制剂。为了获得该信息，T细胞衍生的实验室细胞系和原代T细胞将用超分辨率荧光显微镜成像。各种已知或建议参与HIV用于拆除肌动蛋白进入屏障的信号传导过程的细胞蛋白质将被标记有荧光标签。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粒子跟踪和超分辨率成像技术以高分辨率确定。