CYTOTOXIC LYMPHOCYTE RESPONSES IN ACUTE HIV1 INFECTION

急性期血病毒血症最初下降的原因 人们对HIV-1感染的了解还不完全。HIV-1病毒的出现 特异性细胞毒性淋巴细胞(CTL)在时间上与 病毒血症的下降，在可检测到的中和出现之前 抗体，这表明细胞免疫反应最初可能 包含病毒复制。或者，数学建模 表明病毒血症的下降是预期的结果 一种嗜淋巴细胞病态逆转录病毒的种群动力学 病毒血症的急剧下降是由于感染病毒的人数减少 易感染的活化的CD4细胞。几乎所有关于CTL反应的研究 在原发感染期间已被局限于识别的分析 实验室病毒株的广度和特异性 最初的反应是什么还不清楚。尽管最近的纵向 对单个原发感染者的研究表明，CTL可以 施加免疫压力，序列的出现证明了这一点 主要CTL表位内的变异，CTL或CD8细胞的能力 要实际控制当前初始病毒的复制还没有 已经被证明了。我建议进行详细的分析 HIV-1原发感染患者的抗病毒细胞免疫反应。我们有 开发的体外检测方法，允许直接检查 CD8细胞和CTL克隆的抗病毒活性。我们也在开发 一种适用于快速表达PCR扩增产物的新技术 利用牛痘病毒感染-质粒组合的病毒基因 转染法，以便可以评估对自体病毒的反应 效率很高。利用这些技术，我将进行详细的研究 关键时期CTL和CD8细胞的抗病毒活性 病毒载量下降的初次感染期，依次为 以确定免疫系统对这种下降的贡献。 通过波士顿的一群初次感染者和通过 与加州大学旧金山分校建立了既定的职能合作，我们可以访问 在初次感染期间对人的血液和组织样本进行检测， 在血清转换之前和在建立病毒集之前 指向。这些疫苗的抗病毒免疫反应分析 个人将提供有关相关人员的重要信息 并帮助指导治疗性免疫系统的设计 战略和疫苗。具体地说，我建议：1.描述 自体CTL反应的大小、广度和特异性 初次感染时血液和组织中的病毒，高峰期 病毒血症正在减少。2.检测CD8细胞和CTL的能力 克隆以抑制自体病毒的复制 急性感染期间病毒血症下降。

The factors responsible for the initial fall in viremia during acute HIV-1 infection are incompletely understood. The appearance of HIV-1 specific cytotoxic lymphocytes (CTL) has been temporally correlated with the decline in viremia, before the appearance of detectable neutralizing antibodies, suggesting that the cellular immune response may initially contain viral replication. Alternatively, mathematical modeling suggests that the decline in viremia is the result of expected population dynamics of a lymphotropic cytopathic retrovirus, and that the sharp decline in viremia is a result of diminished numbers of infectable activated CD4 cells. Almost all studies of CTL responses during primary infection have been limited to analysis of recognition of laboratory strains of virus, such that the breadth and specificity of the initial response is not known. Although recent longitudinal studies in a single person with primary infection suggest that CTL can exert immune pressure, as evidenced by the emergence of sequence variation within a dominant CTL epitope, the ability of CTL or CD8 cells to actually control replication of the initial virus present has not been demonstrated. I propose to perform a detailed analysis of the antiviral cellular immune response in primary HIV-1 infection. We have developed in vitro assays which allow for the direct examination of antiviral activity of CD8 cells and CTL clones. We are also developing an adapted novel technique for the rapid expression of PCR amplified virus genes using a combination of vaccinia infection-plasmid transfection, such that responses to autologous virus can be assessed efficiently. Using these techniques, I will conduct a detailed study of the antiviral activity of CTL and CD8 cells during the critical period of primary infection when the viral load is declining, in order to determine the contribution of the immune system to this decline. Through a cohort of persons with primary infection in Boston and through an established and functional collaboration with UCSF, we have access to blood and tissue specimens from persons during primary infection, prior to seroconversion and prior to the establishment of the viral set point. An analysis of the antiviral immune response in these individuals will provide important information regarding the correlates of protective immunity and help guide the design of therapeutic strategies and vaccines. Specifically, I propose to: 1. Characterize the magnitude, breadth and specificity of the CTL response to autologous virus in blood and tissues at the time of primary infection, when peak viremia is declining. 2. Determine the ability of CD8 cells and CTL clones to inhibit replication of autologous virus at the time of declining viremia during acute infection.