STUDIES OF HIV-1 ISOLATES AND SEQUENCES IN WOMEN

病毒学和免疫学因素，包括HIV-1病毒载量、序列 多样性、病毒表型和中和抗体滴度， 与控制HIV-1感染和疾病进展有关。 我们 建议研究这些因素在CD 4耗竭中的作用， 确定发生的病毒学和免疫学事件的时间进程 随着对HIV-1感染的控制减弱和疾病进展。 我们将 分析布朗克斯黎巴嫩的女性血液样本 - 山 西奈-贝斯以色列WIHS队列，这将反映种族和 美国女性HIV-1感染的种族分布。 妇女将 被选中的人显示广泛的CD 4计数，记录模式 CD 4细胞下降和临床状态。 我们计划进行集中的病毒学、分子学和免疫学分析 从这些特征鲜明的女性身上挑选出的系列样本。 我们的目标 目的：1）确定主要中和抗体的核苷酸序列， HIV-1包膜基因（V3环）的决定子，以测量 每个时间点存在的序列多样性程度; 2）定量 外周血单个核细胞体外培养HIV-1 稀释度与确定多样性之间的关系 和单个患者样本内的病毒载量; 3）确定病毒 系列HIV-1分离株的表型;以及4）研究 个体HIV-1准种通过测定自体血清。 我们将 检查在两种栽培中单个准种的中和作用 病毒和插入感染性病毒的V3环序列的分子克隆 矢量，并应开始表征的性质， 逃避中和的病毒的逃逸。 然后我们将 将分子、病毒和中和数据与 疾病进展和CD 4耗竭。 这些分析旨在 在阐明HIV-1发病机制，感染控制， 疫苗设计;他们还可以识别信号HIV-1的病毒学事件 疾病进展，并作为即将发生的临床 感染者的恶化。

Virologic and immunologic factors including HIV-1 viral load, sequence diversity, virus phenotype, and titer of neutralizing antibody are relevant to the control of HIV-1 infection and disease progression. We propose to examine the role of these factors in CD4 depletion and to determine the time course of virologic and immunologic events that occur as control of HIV-1 infection diminishes and disease progresses. We will analyze serial blood specimens from selected women in the Bronx Lebanon - Mt. Sinai - Beth Israel WIHS cohort, which will reflect the racial and ethnic distribution of HIV-1 infection in women in the US. Women will be selected who display a broad range of CD4 counts, documented patterns of CD4 cell decline, and clinical status. We plan to perform focused virologic, molecular, and immunologic analyses on selected serial samples from these well-characterized women. We aim to: 1) Determine the nucleotide sequence of the principal neutralizing determinant of the HIV-1 envelope gene (V3 loop) in order to measure the degree of sequence diversity present at each time point; 2) quantitate HIV-1 cultured from peripheral blood mononuclear cells by limiting dilution and determine the relationship between the degree of diversity and viral load within single patient specimens; 3) determine the viral phenotype of serial HIV-1 isolates; and 4) study neutralization of individual HIV-1 quasispecies by assaying autologous sera. We shall examine neutralization of individual quasispecies in both cultivated virus and molecular clones of V3 loop sequences inserted into infectious vectors and shall begin to characterize the nature of the neutralization escape of the viruses which escape neutralization. We then will correlate the molecular, viral, and neutralization data with the pace of disease progression and CD4 depletion in women. These analyses are aimed at elucidating issues in HIV-1 pathogenesis, control of infection, and vaccine design; they also may identify virologic events that signal HIV-1 disease progression and serve as useful markers of impending clinical deterioration in infected individuals.