CD4+T CELL EXPANSIONS IN HIV

本申请建议使用从两个ACTG中收集的PBMC标本， 回顾性研究CD 4细胞克隆扩增的方案 HIV感染中出现的淋巴细胞库及其反应 到HAART。 PI最近发现了大型克隆/寡克隆 在有限数量的HIV患者中，CD 4细胞扩增。 PI先前的工作表明，CD 4细胞的克隆扩增 在147名健康人的大样本中没有发现 儿童和成人，从广泛的年龄范围，或在自身免疫性 疾病状态，如类风湿性关节炎。 这样的克隆 CD 4细胞扩增可能是HIV感染所特有的。 据推测 扩增的CD 4细胞克隆是由增殖到特定的 HIV抗原，但它们已被赋予功能缺陷， 疾病过程。 这项提案的目标是确定克隆 或表达特定TCR的CD 4细胞亚群的寡克隆性质 HIV患者中的V-β决定因素，治疗前和随访 到80周的HAART 代表扩展和非扩展的子集 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，和3）导出待在一个或多个细胞中表达的TCR序列， 转基因杂交瘤细胞模型用于筛选对HIV的特异性反应性 抗原表位 此外，将进行实验以确定 扩增的CD 4细胞亚群是否携带增加的HIV DNA载量， 与非扩展子集的比较，以及是否存在任何差异水平 在延长的HAART后病毒DNA中持续存在。

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.