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利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 为了全面评价恒河猴巨细胞病毒特异性细胞免疫功能，采用干扰素-γELISPOT、细胞内细胞因子染色和四聚体等多种技术对恒河猴巨细胞病毒免疫优势CD8+和CD4+T细胞表位进行了研究。已确定了4个MAMU-A*01限制性表位，并合成了它们的四聚体。此外，还构建了表达恒河猴CMV即刻早期1和2蛋白、pp65蛋白和IL-10蛋白的疫苗重组体，并合成了15个氨基酸重叠的11肽。到目前为止的结果表明，自然感染CMV的猕猴具有针对猕猴CMV即刻早期蛋白表位的循环CD8+T淋巴细胞的高频率(高达6%)。利用重叠多肽对免疫优势表位进行映射，发现在即刻早期和pp65基因中有几个新的CMV表位。这些表位的MHC I类限制正在确定中。这些研究将能够更准确地评估CMV特异性细胞免疫的保护性相关性，并有助于设计适当的疫苗策略来预防CMV疾病。与艾滋病有关。

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+ and CD4+ 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+ T lymphocytes (as high as 6%) 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.