Minimal Levels of HIV-1 in Vaccinated & Exposed Persons

为了超出所提供的空间，我们在1997年底启动了一项研究，以确定血清阴性者中报告多次与感染艾滋病毒的伴侣发生高危性接触的艾滋病毒-1感染的最低水平，称为暴露的血清阴性。在10名明确的ES患者中，可检测到HIV-L特异性细胞毒性r淋巴细胞(CTL)，我们在两个ES(附录A)中发现了HIV-1感染，水平极低(范围：每百万个细胞0.1-0.01拷贝)，远低于常规检测的检测下限。使用相同的方法，我们确定了SIV感染水平低的猕猴的平行证据，这些猕猴的病毒感染是短暂的病毒感染或通过常规检测获得疫苗保护的。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，这些人按常规检测为一过性病毒携带者。这些发现表明，只有通过数百或数千次聚合酶链式反应扩增和对大量细胞进行序列分析的改进技术，才能在ES和接种疫苗的人中证明最低水平的艾滋病毒-1感染(MLHI)。在这里，我们建议扩展我们的初步结果，使过程机械化，并在我们的ES队列和NIH资助的疫苗试验(主要是HIV疫苗试验网络(HVTN))的疫苗接受者中更严格地评估MLHI。我们还将利用从我们的检测中分离出的病毒或序列的可用性来表征突破性MLHI感染中病毒的基因型，并将这些特征与疫苗毒株进行比较。具体目标是：1.改进方法学，以确定疫苗试验参与者和ES个人中短暂或持久的最低水平的HIV-1感染。我们将确定传统病毒学、血清学或T细胞介导的免疫学检测表明可能患有一过性感染的人中HIV-1的频率和复制状态。2.确定MLHI人群外周血和组织中存在的HIV-1毒株的基因型，并确定接种疫苗或反复暴露于HIV-1是否对HIV-1感染的最低水平产生影响。这些研究应有助于确定减轻和/或实质控制HIV-1感染所需的病毒学和免疫学特征，这是艾滋病疫苗开发的关键目标。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED, 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. PERFORMANCE SITE ========================================Section End===========================================