Persistent Reservoirs of HIV-1 Infection

HIV-1 感染的持续储存

• 批准号: 6882086
• 负责人: DANIEL SETH FIERER
• 金额: $ 14.81万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882086
• 关键词: HIV infections; antiviral agents; cell sorting; clinical research; disease reservoirs; flow cytometry; human therapy evaluation; immunotherapy; lymphocyte; monocyte; nucleic acid sequence; patient oriented research; polymerase chain reaction; provirus; virus DNA; virus load; virus replication

DESCRIPTION (Provided by applicant): Despite long-term suppression of HIV-1 viral load in plasma to undetectable levels, HIV-1 has not been eradicated from patients treated with potent antiretroviral regimens. The available evidence suggests this inability to eradicate HIV-1 infection is due to ongoing low-level viral replication and persistent reservoirs of infection despite potent therapy. We posit that extra-chromosomal circular HIV DNA (DNA circles), which is detectable in peripheral blood monocytic cells (PBMC) despite long-term suppression of HIV viral load in plasma to undetectable levels, is a i marker for continued viral replication in these patients and may demonstrate the source(s) of persistent viral replication and potential persistent reservoirs of infection. In this application, we propose 3 studies to test these hypotheses and to try to answer these basic questions about HIV-1 pathogenesis: 1) Evolution of HIV sequences occurs during HIV-1 replication and so is a marker for ongoing viral replication. We will evaluate for sequence evolution over time between HIV DNA circles (representing new cellular infection) and integrated proviral DNA (archival sequence) in PBMC from patients who have had long-term suppression of viral replication with potent antiretroviral regimens; 2) Despite long-term suppression of plasma HIV-1 viral load to undetectable levels with potent antiretroviral medication, upon discontinuation of the medication, plasma viremia quickly returns. The exact source of this "rebounding" virus is not known, however. We will therefore determine the genetic relatedness between the low-level replicating virus found in PBMC during long-term antiretroviral therapy, which we posit is the main source of the "rebounding" virus (represented by DNA circles) versus the virus that rebounds after discontinuation of antiretroviral therapy, and further, in comparison with proviral DNA sequences found in seminal mononuclear cells, a putative persistent reservoir of infection; and 3) The sub-population(s) of lymphocytes that are infected during ongoing low-level HIV-1 replication in patients receiving potent antiretroviral therapy have not been identified. Identification of these newly infected lymphocytes would supply clues as to the origin of the persistent reservoirs of infection in these patients. We will therefore combine cell sorting techniques with our assay for DNA circles to identify specific sub-population(s) within PBMC and seminal mononuclear cells that harbor DNA circles. Identification of the reservoirs of persistent HIV-1 infection is crucial for understanding the mechanisms of persistence of HIV-1 infection in patients treated with potent antiretroviral medications and may lead to the development of new therapeutic agents and approaches for the eradication of HIV-1 from infected patients. These studies will also provide me with the opportunity to develop the required skills to reach my long-term goal: a career as an independent investigator and academician performing patient-oriented translational research.

描述（由申请人提供）：尽管长期抑制了HIV-1 血浆中的病毒载荷至无法检测到的水平，HIV-1尚未从中消除 接受有效抗逆转录病毒方案治疗的患者。可用证据 表明这种无法消除HIV-1感染是由于持续的 尽管 有效的疗法。我们认为该外染色体圆形HIV DNA（DNA圆）， 尽管在外周血单核细胞（PBMC）中可检测 长期抑制血浆中HIV病毒载量至无法检测到的水平，是一个 我标记了这些患者的持续病毒复制，并可能证明 持续病毒复制和潜在持久性的来源 感染的水库。在此应用中，我们提出了3项研究以测试 这些假设，并试图回答有关HIV-1的这些基本问题 发病机理：1）HIV序列的进化发生在HIV-1复制期间 因此，持续的病毒复制的标志也是如此。我们将评估顺序 HIV DNA圆圈之间的演变（代表新的细胞 感染）和PBMC中的综合病毒DNA（档案序列） 长期抑制病毒复制的患者 抗逆转录病毒方案； 2）尽管长期抑制了血浆HIV-1病毒 通过有效的抗逆转录病毒药物的负载到无法检测到的水平， 停用药物，血浆病毒血症很快就会恢复。确切的 但是，这种“反弹”病毒的来源尚不清楚。因此，我们会 确定发现的低级复制病毒之间的遗传相关性 在长期抗逆转录病毒疗法期间的PBMC中，我们认为这是主要的 “反弹”病毒的来源（由DNA圈代表）与病毒 在中止抗逆转录病毒疗法后，反弹，进一步 与在精液单核细胞中发现的病毒DNA序列的比较，A 推定的持续感染库； 3） 在持续的低级HIV-1复制中感染的淋巴细胞 尚未确定接受有效抗逆转录病毒疗法的患者。 鉴定这些新感染的淋巴细胞将提供有关 这些患者的持续感染库的起源。我们将 因此，将细胞分选技术与我们对DNA圆的测定 确定PBMC和精确单核细胞中的特定亚群 那个藏有DNA圈子。 鉴定持续性HIV-1感染的储层对于 了解患者HIV-1感染持续性的机制 用有效的抗逆转录病毒药物治疗，并可能导致发展 从 感染的患者。这些研究还将为我提供机会 发展所需的技能以实现我的长期目标：作为一个职业 独立的研究员和院士表演以患者为导向的 翻译研究。