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

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。为了全面评估恒河猴CMV特异性细胞免疫，利用多种技术绘制恒河猴CMV免疫优势CD8+和CD4+ T淋巴细胞表位，包括干扰素- γ ELISPOT测定、细胞内细胞因子染色测定和四聚体。鉴定出4个Mamu-A*01限制性表位并合成了它们的四聚体。此外，还构建了表达恒河巨细胞病毒早期1和2蛋白、pp65蛋白和IL-10蛋白的痘苗重组体，并合成了跨越这些巨河巨细胞病毒蛋白的15个aa重叠11个肽段。迄今为止的结果表明，自然感染CMV的繁殖恒河猴具有高频率的循环CD8+ T淋巴细胞（高达6%）靶向恒河猴CMV即时早期蛋白的表位。利用重叠肽对免疫显性表位进行定位，揭示了直接早期和pp65基因中几个新的CMV表位。正在确定这些表位的MHC I类限制。这些研究将能够更精确地评估巨细胞病毒特异性细胞免疫的保护性相关因素，并有助于设计预防巨细胞病毒疾病的适当疫苗策略。

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 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.