CD4+T CELL EXPANSIONS IN HIV

此应用程序建议使用从两个ACTG收集的PBMC样本 回顾研究CD_4细胞克隆扩张的方案 HIV感染中出现的淋巴细胞库及其反应(S) 敬哈尔特。PI最近发现了大克隆/寡克隆 在数量有限的HIV患者中，CD4细胞的扩张。 PI之前的工作表明，CD4细胞的克隆性扩张 在147名健康人的大样本中没有发现 儿童和成人，来自广泛的年龄段，或处于自身免疫状态 疾病状态，如类风湿性关节炎。因此，这样的克隆 CD4细胞的扩张可能是HIV感染所特有的。这是假设的 扩增的CD4细胞克隆从增殖到特异性 HIV抗原，但由于 疾病过程。这项提议的目标是确定克隆 或表达特定TCR的CD4细胞亚群的寡克隆性 HIV患者治疗前和随访中的V-β决定因素 到80周的HAART。表示扩展的和非扩展的子集 将分析扩展的V-β表型以：1)比较它们的 体外增殖能力，2)确定TCR多样性的变化 在HAART之后，以及3)导出TCR序列以在 用于筛选HIV特异性反应性的转基因杂交瘤细胞模型 抗原表位。此外，还将进行实验，以确定 扩增的CD4细胞亚群是否含有增加的HIV DNA 与非扩展子集的比较，以及是否存在差异水平 在延长HAART后，病毒DNA中的DNA保持不变。

This application proposes to use PBMC specimens collected from two ACTG protocols to retrospectively study clonal expansions in the CD4 lymphocyte repertoire that occur in HIV infection and their response(s) to HAART. The PI has recently identified large clonal/oligoclonal expansions in CD4 cells in a limited number of patients with HIV. Previous work by the PI has shown that clonal expansions of CD4 cells in the periphery are not found in a large sampling of 147 healthy children and adults, from a broad range of ages, or in autoimmune disease states, such as rheumatoid arthritis. Therefore, such clonal CD4 cell expansions may be unique to HIV infection. It is hypothesized that the expanded CD4 cell clones arise from proliferation to specific HIV antigens, but they have been rendered functionally deficient by the disease process. The goal of this proposal is to determine the clonal or oligoclonal nature of CD4 cell subsets that express particular TCR V-beta determinants in HIV patients, prior to therapy and following up to 80 weeks of HAART. Subsets representing both expanded and non- expanded V-beta phenotypes will be analyzed to: 1) compare their proliferative capacity in vitro, 2) determine changes in TCR diversity following HAART, and 3) derive TCR sequences to be expressed in a transgenic hybridoma cell model to screen for specific reactivity to HIV antigen epitopes. In addition, experiments will be done to determine whether expanded subsets of CD4 cells harbor increased HIV DNA load in comparison to non-expanded subsets, and whether any differential levels in viral DNA persist following extended HAART.