MAPPING OF IMMUNODOMINANT CD8+ AND CD4+ T LYMPHOCYTE EPITOPES IN RHESUS CMV

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 为了对恒河猴CMV特异性细胞免疫进行全面评价，正在使用多种技术，包括干扰素-γ ELISPOT测定、细胞内细胞因子染色测定和四聚体，对恒河猴CMV中的免疫显性CD 8阳性和CD 4阳性T淋巴细胞表位进行定位。 已经鉴定了四个Mamu-A*01限制性表位，并合成了它们的四聚体。 此外，已经构建了表达恒河猴CMV立即早期1和2蛋白、pp 65蛋白和IL-10蛋白的牛痘重组体，并且已经合成了跨越这些恒河猴CMV蛋白的15个氨基酸重叠的11个肽。 迄今为止的结果表明，自然CMV感染的种猴具有高频率的循环CD 8阳性T淋巴细胞（高达6%）靶向恒河猴CMV立即早期蛋白表位。 使用重叠肽的免疫显性表位的映射揭示了几个新的CMV表位在立即早期和pp 65基因。 这些表位的MHC I类限制正在确定中。 这些研究将能够更精确地评估CMV特异性细胞免疫的保护相关性，并有助于设计预防CMV疾病的适当疫苗策略。艾滋病相关

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In order to conduct a comprehensive evaluation of CMV-specific cellular immunity in rhesus macaques, immunodominant CD8-positive and CD4-positive T lymphocyte epitopes in rhesus CMV are being mapped using a variety of techniques including interferon-gamma ELISPOT assays, intracellular cytokine staining assay and tetramers. Four Mamu-A*01 restricted epitopes have been identified and their tetramers synthesized. In addition vaccinia recombinants expressing the rhesus CMV immediate early 1 and 2 proteins, pp65 protein and IL-10 protein have been constructed, and 15 aa overlapping 11 peptides spanning these rhesus CMV proteins have been synthesized. Results to date show that naturally CMV-infected breeder rhesus macaques have high frequencies of circulating CD8-positive T lymphocytes (as high as 6 percent) targeting epitopes in the rhesus CMV immediate early proteins. Mapping of immunodominant epitopes using overlapping peptides has revealed several new CMV epitopes in the immediate early and pp65 genes. The MHC Class I restriction of these epitopes is being determined. These studies will enable more precise assessment of the protective correlates of CMV-specific cellular immunity and help in the design of appropriate vaccine strategies for prevention of CMV disease. AIDS related.