Minimal Levels of HIV-1 in Vaccinated & Exposed Persons

在1997年底，我们开始了一项研究，以确定血清反应阴性者的最低艾滋病毒-1感染水平 报告重复与HIV-1感染者发生高危性接触的人，称为暴露者， 血清阴性（ES）。在10个明确定义的ES个体中，可检测到HIV-1特异性细胞毒性r- 淋巴细胞（CTL）反应，我们证明了在两个ES中存在HIV-1感染（附录A）， 非常低的水平（范围：0.1 - 0.01拷贝/百万细胞），远低于 常规测定。使用相同的方法，我们发现了低水平SIV的平行证据， 在通过常规检测获得短暂病毒血症或疫苗保护的猕猴中的感染。最 最近，我们在三名接种疫苗的人中发现了持续的低水平HIV-1感染， 通过常规测定法检测为一过性病毒血症。这些发现表明，最低水平的HIV-1感染 （MLHI）可以在ES和接种疫苗的人中证明，只有通过改进技术， 对大量细胞进行数千次PCR扩增和序列分析。在此，我们建议 扩展我们的初步结果，使过程机械化，并更严格地评估MLHI， 我们的ES和疫苗接受者队列来自NIH资助的疫苗试验，主要是HIV疫苗试验 网络（HVTN）。我们还将利用从我们的研究中分离的病毒或序列的可用性。 分析以表征突破性MLHI感染中的病毒基因型，并比较这些 疫苗株的特性。具体目标是：1。改进识别瞬态的方法 或在疫苗试验参与者和ES个体中持续最低水平的HIV-1感染。我们将 确定可能有短暂感染的人中HIV-1的频率和复制状态， 通过常规的病毒学或血清学或T细胞介导的免疫学测定来指示。2.表征 MLHI患者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对HIV-1最低水平的减弱有影响 感染 这些研究应有助于定义所需的病毒学和免疫学特征， 减弱和/或基本上控制HIV-1感染，这是AIDS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.