Minimal Levels of HIV-1 in Vaccinated & Exposed Persons

1997年底，我们启动了一项研究，以确定血清阴性者感染hiv-1病毒的最低水平。 世卫组织报告称，反复与感染艾滋病毒的L感染的伴侣发生高危性接触，称为接触 血清阴性。在10名明确的ES患者中，可检测到HIV-L特异性细胞毒r- 淋巴细胞(CTL)反应，我们证明在两个ES(附录A)中存在HIV-1感染 极低的水平(范围：每百万个细胞0.1-0.01个拷贝)，远远低于 常规化验。使用相同的方法，我们确定了低水平SIV的类似证据 通过常规检测获得短暂病毒感染或疫苗保护的猕猴的感染。多数 最近，我们发现在三名接种疫苗的人中，艾滋病毒-1的感染水平持续较低。 用常规的化验方法检测出瞬时病毒。这些发现表明，最低水平的艾滋病毒-1感染 (MLHI)可以在ES和接种疫苗的人身上展示，只有通过数百或 对大量细胞进行数千次聚合酶链式反应扩增和序列分析。在此，我们建议 扩展我们的初步结果，使流程机械化，并更严格地评估MLHI 我们的ES和来自NIH资助的疫苗试验的疫苗接受者的队列，主要是HIV疫苗试验 网络(HVTN)。我们还将利用从我们的 突破性MLHI感染病毒的基因分型方法及其比较 疫苗毒株的特征。具体目标是：1.改进识别瞬变的方法 或在疫苗试验参与者和ES个人中持续最低水平的HIV-1感染。我们会 确定HIV-1在可能有短暂感染的人群中的频率和复制状态 由传统病毒学、血清学或T细胞介导的免疫学检测表明。2.刻画 MLHI患者外周血和组织中HIV-1毒株的基因分型 接种疫苗或反复暴露于HIV-1对HIV-1减弱的最低水平有影响 感染。 这些研究应该有助于定义病毒学和免疫学特征所需的 减少和/或基本上控制艾滋病毒-1感染，这是艾滋病疫苗开发的关键目标。

In late 1997, we initiated a study to identify minimal levels of HIV-1 infection in seronegative persons who reported repeated high-risk sexual exposures to HIV-l-infected partners, termed as exposed seronegative (ES). Among ten well-defined ES individuals with detectable HiV-l-specific cytotoxic r- lymphocytes (CTL) responses, we demonstrated presence of HIV-1 infection in two ES (Appendix A) at extraordinarily low levels (range: 0.1 - 0.01 copies per million cells) that are well below the detection limit of conventional assays. Using same methodologies, we identified parallel evidence for low levels of SIV infection in macaques that were transiently viremic or vaccine-protected by conventional testing. Most recently, we have found persistent low levels of HIV-1 infection in three vaccinated persons who were transiently viremic by conventional assays. These findings indicate that minimal levels of HIV-1 infection (MLHI) can be demonstrated in ES and vaccinated persons, only by improved technologies with hundreds or thousands of PCR amplifications and sequence analyses on large amounts of cells. Here, we propose to extend our preliminary results to mechanize the processes and to assess for MLHI more rigorously among our cohort of ES and vaccine recipients from NIH-funded vaccine trials, mainly the HIV Vaccine Trails Network (HVTN). We will also take advantage of the availability of virus or sequences isolated from our assays to characterize the genotype of virus in breakthrough MLHI infection and compare these characteristics with vaccine strains. The Specific Aims are: 1. To improve methodologies to identify transient or persistent minimal levels of HIV-1 infection in vaccine trial participants and ES individuals. We will ascertain the frequency and replication state of HIV-1 in persons who may have transient infection as indicated by conventional virologic, or serologic, or T cell-mediated immunologic assays. 2. To characterize the genotype of HIV-1 strains present in peripheral blood and tissues in MLHI persons and to determine if there is impact of vaccination or repeated exposures to HIV-1 on the attenuated minimal levels of HIV-1 infection. These studies should assist definition of virologic and immunologic characteristics required to attenuate and/or substantially control HIV-1 infection, a critical goal of AIDS vaccine development.