SIV-specific Mamu-E-restricted CD8+ T cells

描述（由申请人提供）：拟议研究的目的是确定非经典MHC I类分子Mamu-E是否可以将siv衍生肽呈现给印度恒河猴的免疫系统。然后，我们将确定是否可以通过疫苗接种引起对这些肽的反应，以及它们是否可以控制SIV复制。结核分枝杆菌(Mtb)-、巨细胞病毒(CMV)-和伤寒沙门氏菌特异性CD8+ T细胞已被证明能够识别HLA-E结合的肽。此外，小鼠接种了转运蛋白与抗原加工相关（TAP）抑制的树突状细胞（DCs），产生了Qa-1b- （HLA-E的功能同源物）限制性CD8+ T细胞，从而保护小鼠免受多种肿瘤的侵袭。因此，识别HLA-E或其小鼠同源Qa-1b的CD8+ T细胞可能在病原体和肿瘤的免疫监视中发挥关键作用。有趣的是，HLA-E与其他经典的MHC I类分子不同，在非人类灵长类物种中具有很好的保守性。这种程度的保守在这些非人灵长类动物MHC-E分子的肽结合槽中最为明显，这表明MHC-E在人类和非人灵长类动物的肽结合中都起着重要作用。我们在SIV感染的印度恒河猴中进行了初步研究，结果表明，符合HLA-E结合基序的SIV衍生肽在IFN3 ELISPOT检测中被识别。我们正在提出一系列实验来探索SIV特异性Mamu-E限制性CD8+ T细胞在SIV免疫监视中发挥作用的创新假设。我们将从Mamu-E中洗脱肽以确定其肽结合基序，扫描SIVmac239蛋白质组以预测Mamu-E结合物，然后使用IFN3 ELISPOT和PBMC从SIV感染的印度恒河猴中筛选这些肽。然后，我们将培养受活性肽刺激的细胞系，并使用Mamu-E转染剂和Mamu-E四聚体确定它们是否确实受Mamu-E限制。在本研究的第二部分中，我们假设通过疫苗接种产生mamu - e特异性CD8+ T细胞反应将扩大抗病毒反应的范围，并有助于减少致病性异源病毒攻击后的病毒复制。！

DESCRIPTION (provided by applicant): The goal of the proposed research is to determine if the non-classical MHC class I molecule, Mamu-E, can present SIV-derived peptides to the immune system of the Indian rhesus macaque. We would then determine if the responses to these peptides can be elicited by vaccination and if they can control SIV replication. Mycobacterium tuberculosis (Mtb)-, Cytomegalovirus (CMV)- and Salmonella typhi-specific CD8+ T cells have been shown to recognize peptides bound by HLA-E. Furthermore, mice vaccinated with Transporter associated with antigen processing (TAP)-inhibited dendritic cells (DCs) develop Qa-1b- (the functional homologue of HLA-E) restricted CD8+ T cells that confer protection from challenge with a variety of tumors. CD8+ T cells that recognize HLA-E or its murine homologue Qa-1b may, therefore, play a critical role in immunological surveillance of both pathogens and tumors. Interestingly, HLA-E, unlike other classical MHC class I molecules, is well conserved across non-human primate species. This degree of conservation is most pronounced in the peptide-binding groove of these non-human primate MHC-E molecules suggesting that MHC-E plays an important role in binding peptides in both humans and non-human primate species. We have conducted preliminary studies in SIV-infected Indian rhesus macaques and have shown that SIV- derived peptides conforming to the HLA-E binding motif are recognized in IFN3 ELISPOT assays. We are proposing a series of experiments to explore the innovative hypothesis that SIV-specific Mamu-E- restricted CD8+ T cells play a role in immune surveillance against SIV. We will elute peptides from Mamu-E to define its peptide-binding motif, scan the SIVmac239 proteome for predicted Mamu-E binders and then screen these peptides using IFN3 ELISPOT with PBMC from SIV- infected Indian rhesus macaques. We will then grow cell lines that are stimulated by the reactive peptides and determine whether they are indeed Mamu-E-restricted using Mamu-E transfectants and Mamu-E tetramers. In the second part of our application we hypothesize that the generation of Mamu-E-specific CD8+ T cell responses by vaccination will broaden the repertoire of antiviral responses and aid in reducing viral replication after challenge with a pathogenic heterologous virus. ! PUBLIC HEALTH RELEVANCE: Finding a vaccine for HIV is probably one of the most important goals of biomedical research. We will use approaches derived from cancer research to try to find new targets for an HIV vaccine. !