HIV-1 evolution in the female genital tract and trafficking to the blood

HIV-1 在女性生殖道中的进化和贩运到血液中

• 批准号: 8081383
• 负责人: Lisa M Frenkel
• 金额: $ 1.76万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-21 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8081383
• 关键词: Address; Biopsy; Blood; Cell Line; Cell Proliferation; Cells; Cervical; Cervix Uteri; Clonal Expansion; Coculture Techniques; Coitus; DNA; Detection; Drug resistance; Evolution; Female; Frequencies; Genital system; Genome; HIV; HIV Infections; HIV-1; Half-Life; Individual; Infection; Infection prevention; Intervention; Length; Longevity; Mediating; Minority; Modeling; Mucous Membrane; Mutation; Nucleosides; Phylogenetic Analysis; Plasma; Population; Production; Proliferating; Proviruses; Reading Frames; Role; Shapes; Site; Specimen; Sputum; Tampons; Time; Tissues; Trauma; Variant; Viral; Viral Genome; Viral Load result; Virion; Virus; Virus Replication; Visit; Woman; antiretroviral therapy; cell type; innovation; non-nucleoside reverse transcriptase inhibitors; novel; novel strategies; trafficking

DESCRIPTION (provided by applicant): In the mid-1990's, modeling of viral decay predicted that antiretroviral therapy (ART) would result in clearance of HIV infection with ~3 years of treatment. However, HIV was found to persist during suppressive ART. The half-life of infection was estimated to extend for many years, such that viral clearance would not be expected to occur during a normal life span. Our and others' studies of HIV that persists during suppressive ART have observed low-level viral replication in a minority (~20%) of individuals. We have studied HIV persisting in blood, sputum and genital tissues, all of which could serve as potential viral reservoirs during effective ART. Utilizing single genome amplification (SGA), we independently-derived and directly sequenced (to avoid detection of PCR-mediated mutations) sufficient viral templates from multiple specimens to characterize viral populations and their changes during suppressive ART. We detected sizable populations of identical viral genomes, here termed monotypic virus, in various cell types. SGA of low-level plasma viral blips (<500c/mL) revealed monotypic viruses in individuals that phylogenetic and drug resistance analyses showed no evidence of viral replication. We reason that blips of monotypic virus could result from production of virions without full- cycles of replication, as nucleoside- and non-nucleoside reverse transcriptase inhibitors (NRTI and NNRTI) would prevent infection of additional cells. As estimates of viral burst size suggest that a single HIV infected cell could not produce sufficient virions to generate a viral load of 50-500c/mL, this gave further support to a developing hypothesis that HIV proviruses are amplified in proliferating cells, and that at times these cells produce virions without full rounds of infection. If our hypothesis is correct, cellular proliferation could be an important mechanism that over time perpetuates HIV infection, a concept that has not been previously characterized. The hypotheses of this proposal are that: (1) Clonal expansion (i.e., proliferation) of HIV infected cells results in multiple cells, each with an identical viral sequence integrated at the same site in the host's genome. (2) A substantial proportion of viruses detected in the female genital mucosa and blood are derived from the proliferation of HIV infected cells, and these viruses become more prominent during suppressive ART. (3) Monotypic proviral sequences include templates that are replication competent. We Aim to: 1: Determine if monotypic (identical) HIV env sequences in the uterine cervix result from proliferation of infected cells or from bursts of viral replication 2: Determine if whole monotypic HIV genomes include replication competent viruses Our proposed studies exploring the role of cellular proliferation in perpetuating HIV infection could alter current paradigms explaining viral persistence and shape interventions needed to cure HIV. PUBLIC HEALTH RELEVANCE: We have observed that HIV populations may grow in the genital tract because cells with virus appear to proliferate, probably due to infections and traumas (e.g., sexual intercourse, tampons, etc.). If a significant fraction of genital tract HIV is derived from proliferation of infected cells, this phenomenon may present a major obstacle to curing HIV infection; unless novel approaches can be developed to eliminate HIV infection from the body.

描述（由申请人提供）：在 20 世纪 90 年代中期，病毒衰变模型预测抗逆转录病毒疗法 (ART) 将通过约 3 年的治疗清除 HIV 感染。然而，在抑制性抗逆转录病毒治疗期间，艾滋病毒被发现持续存在。据估计，感染的半衰期会延长很多年，因此在正常寿命期间不会发生病毒清除。我们和其他人对抑制性 ART 期间持续存在的 HIV 的研究观察到少数（约 20%）个体中病毒复制水平较低。我们研究了血液、痰液和生殖器组织中持续存在的艾滋病毒，所有这些都可以在有效的抗逆转录病毒治疗期间作为潜在的病毒储存库。利用单基因组扩增 (SGA)，我们从多个样本中独立衍生并直接测序（以避免检测 PCR 介导的突变）足够的病毒模板，以表征病毒群体及其在抑制性 ART 期间的变化。我们在不同的细胞类型中检测到大量相同的病毒基因组，这里称为单型病毒。低水平血浆病毒 blip (<500c/mL) 的 SGA 显示个体中存在单型病毒，系统发育和耐药性分析显示没有病毒复制的证据。我们推断，单型病毒的斑点可能是由于没有完整复制周期的病毒颗粒的产生而导致的，因为核苷和非核苷逆转录酶抑制剂（NRTI 和 NNRTI）会阻止其他细胞的感染。由于对病毒爆发大小的估计表明，单个 HIV 感染细胞无法产生足够的病毒粒子来产生 50-500c/mL 的病毒载量，这进一步支持了一个发展中的假设，即 HIV 前病毒在增殖细胞中扩增，并且在这些细胞在没有进行全轮感染的情况下产生病毒粒子。如果我们的假设是正确的，细胞增殖可能是随着时间的推移使艾滋病毒感染永久存在的重要机制，这一概念以前从未被描述过。该提议的假设是：(1) HIV 感染细胞的克隆扩增（即增殖）导致多个细胞，每个细胞都具有整合在宿主基因组中同一位点的相同病毒序列。 (2)女性生殖器粘膜和血液中检测到的病毒有很大一部分来自HIV感染细胞的增殖，这些病毒在抑制性ART期间变得更加突出。 (3)单型原病毒序列包括具有复制能力的模板。我们的目标是： 1：确定子宫颈中的单型（相同）HIV env 序列是否是受感染细胞增殖或病毒复制爆发的结果 2：确定整个单型 HIV 基因组是否包含有复制能力的病毒 我们提出的研究探索了使艾滋病毒感染永久化的细胞增殖可能会改变当前解释病毒持久性的范式，并形成治愈艾滋病毒所需的干预措施。 公共卫生相关性：我们观察到，艾滋病毒群体可能会在生殖道中增长，因为带有病毒的细胞似乎会增殖，这可能是由于感染和创伤（例如性交、卫生棉条等）造成的。如果生殖道艾滋病毒的很大一部分来自受感染细胞的增殖，这种现象可能会成为治愈艾滋病毒感染的主要障碍；除非能够开发出新的方法来消除体内的艾滋病毒感染。