HIV Restriction in CD4+ T cells: Host and Viral Factors

描述（由申请人提供）：在我的R 01中，我提议确定HIV是否可以整合到静息的CD 4 + T细胞中，并研究T细胞的活化如何影响该过程。我证明了，与教条相反，整合确实发生在静息细胞中。这项工作导致了HIV潜伏期的体外模型，并形成了HIV储库形成的新范例的基础-储库可以在没有T细胞活化的情况下形成。作为这些研究的一部分，我开发了一种新的，定量的，高灵敏度的HIV整合检测方法。该测定是第一个足够灵敏以测量来自患者的PBMC中的整合的测定。先前的整合测定缺乏必要的灵敏度，因此我的测定是一个显著的进步。在这里，我建议使用新的检测方法来测量体内CD 4+细胞亚群的整合（目的1）。比较HAART患者与HAART患者的整合水平可能提供另一种评估治疗疗效的方法。 在我的R 01研究过程中，我还对HIV的静息T细胞限制产生了兴趣和专业知识。在这里，我建议研究HIV在CD 4+细胞中的限制，检查细胞和病毒因素。我的整合分析对于高度限制的细胞来说足够灵敏。使用我在R 01研究中开发的工具，我将描述血液来源的CD 4+细胞中HIV感染的限制（目标2），并检查辅助病毒蛋白在克服静息T细胞中的限制中所起的作用（目标3）。识别参与限制HIV感染的细胞途径和病毒蛋白质可以为抗逆转录病毒疗法提供新的靶点。 我的长期目标是成为一名独立的医生科学家，从事与临床相关的基础研究。我的短期目标是获得这个奖项，因为它将保证我有75%的受保护时间，我需要这些时间来专注于我的研究并为该领域做出重大贡献。自从成为助理教授以来，我是五本出版物的第一或资深作者。我最近招收了一名本科生，两名研究生和一名博士后，与他们一起投稿，因此我的工作效率提高了。在艾滋病毒研究中发展自己的利基并获得资金（6笔赠款，包括R 01）的能力证明了我作为一名独立的医生科学家取得成功的潜力。最后，来自我的系主任、部门负责人、导师和合作者的推荐信传达了我在宾夕法尼亚大学得到的大力支持。

DESCRIPTION (provided by applicant): In my R01, I proposed to determine if HIV could integrate in resting CD4+ T cells and to study how activation of T cells affects that process. I showed that, contrary to dogma, integration does occur in resting cells. This work led to an in vitro model of HIV latency and formed the basis for a new paradigm of HIV reservoir formation - that reservoirs may form without T cell activation. As part of these studies, I developed a new, quantitative, highly sensitive assay for HIV integration. This assay is the first that is sensitive enough to measure integration in PBMCs from patients. Prior integration assays lack the necessary sensitivity, thus my assay is a significant advance. Here, I propose to use the new assay to measure integration in subsets of CD4+ cells in vivo (Aim 1). Comparing integration levels in patients off HAART to patients on HAART may provide another way to evaluate the efficacy of therapy. In the process of my R01 research, I also developed an interest and expertise in resting T cell restriction of HIV. Here, I propose to study the restriction of HIV in CD4+ cells, examining both the cellular and viral factors. My integration assay is sensitive enough for use in highly restricted cells. Using the tools that I developed during my R01 studies, I will characterize the restrictions to HIV infection in blood-derived CD4+ cells (Aim 2), and examine the role that auxiliary viral proteins play in overcoming the restriction in resting T cells (Aim 3). Identification of cellular pathways and viral proteins involved in restriction of HIV infection could provide new targets for antiretroviral therapies. My long-term goal is to be an independent physician-scientist who conducts basic research that has clinical relevance. My short-term goal is to obtain this award, because it will guarantee me 75% protected time, which I need in order to focus on my research and make significant contributions to the field. Since becoming an Assistant Professor, I am first or senior author on five publications. I have recently recruited one undergraduate student, two graduate students and a postdoc, with whom I have submitted manuscripts, thus my productivity is increasing. The ability to develop my own niche in HIV research and to obtain funding (6 grants, including an R01) demonstrate my potential for success as an independent physician-scientist. Finally, the letters of recommendation from my department chair, division chief, mentor, and collaborators convey the strong support that I receive at Penn.