Characterization of HIV infection in resting CD4 T-cells

静止的CD4 t细胞对生产性HIV感染具有抵抗力，但在疾病进展过程中大量耗尽，是HIV感染者体内病毒的主要储存库。该合作和跨学科研究计划的第二个资助期的目标是揭示和表征静止CD4 t细胞中HIV复制和细胞死亡的障碍和调节因子。基于第一个资助期的激动人心的结果，以及新建立的高效基因敲除（KO）方案和优化的静息CD4 t细胞培养条件，我们现在的目标是在四个工作包中：(i)对这种原代细胞类型中HIV-1感染的宿主依赖因子进行基本表征，（ii）从基因上评估新型细胞Vpx相互作用物的效力-通过蛋白质组学方法鉴定-用于HIV进入后早期阶段。（iii）描述参与限制和感知HIV的假定细胞因子的特征和功能相关性，以及（iv）将KO方法应用于扁桃体组织培养离体模型，以消除复杂淋巴组织中HIV-1感染期间至关重要的免疫耗竭的传感机制和信号通路。总的来说，这些研究将增加我们对HIV-1与静止CD4 t细胞多方面相互作用的分子和病理生理学理解。

Resting CD4 T-cells are resistant to productive HIV infection, yet are massively depleted during disease progression and represent a major reservoir for the virus in HIV-infected individuals. The goal of the second funding period of this collaborative and interdisciplinary research proposal is to unveil and characterize barriers to and regulators of HIV replication and cell death in resting CD4 T-cells. Building on exciting results from the first funding period and newly established protocols for efficient gene knockouts (KO) and optimized cultivation conditions in resting CD4 T-cells, we now aim in four work packages to (i) perform a fundamental characterization of host dependency factors for HIV-1 infection in this primary cell type, (ii) genetically evaluate the potency of novel cellular Vpx interactors - identified by proteomic approaches - for the early post-entry phase of HIV, (iii) characterize the profile and functional relevance of putative cellular factors involved in restriction and sensing of HIV, and (iv) adapt the KO approach to the tonsil histoculture ex vivo model to dismantle sensing machineries and signaling pathways that critically contribute to immunodepletion during HIV-1 infection in complex lymphoid tissues. Collectively, these studies will increase our molecular and pathophysiological understanding of the multi-faceted interactions of HIV-1 with resting CD4 T-cells.