MHC TYPING OF MACAQUES USED IN AIDS RESEARCH

描述(改编自申请人的摘要)：增加的效用 HIV疫苗研发中的恒河猴动物模型和 发病机制研究需要发展参考MHC分型 这一物种的实验室。在这份提案中，申请者计划 建立两个恒河猴MHC分型参考实验室。两者都有 在这些实验室中，将由拥有超过 有十年分析恒河猴MHC的经验。其中之一 这些实验室将在WRPRC的David博士的指导下进行 沃特金斯和其他人将在生物医学灵长类研究中心工作 在罗纳德·邦特罗普博士的指导下，中心(荷兰)。这个 申请者计划同时向北美和欧洲提供服务 科学界对猕猴进行MHC分型。最初，这将是 包括编码结合MHC I类分子的等位基因的PCR-SSP测试 来源于SIV和SIV的多肽。申请者还将提供PCR-SSP 对类似编码MHC II类分子的MHC II类等位基因进行分型 结合来自SIV或SIV的多肽。调查人员建议开发一种 用于分析猕猴MHC第I类和第I类的其他分子技术 II类等位基因。这些将包括参考链构象分析 (RSCA)和恒河猴MHC类II等位基因的直接测序。 调查人员计划利用一家公司(PFI)的资源 开发和销售这些用于分析人类MHC的技术 等位基因。因此，申请人提议向PFI分包合同，以提供 建立适用于恒河猴的RSCA。此外， 申请者将为希望设置MHC打字的个人提供培训 在他们自己的实验室里。最后，申请者将有一组井- 特征化的、MHC定义的细胞系，对分析将是无价的 恒河猴体内的MHC。其中包括一组恒河猴 研究人员已知的猕猴B淋巴母细胞系(BLCL) 所有表达的MHC I类和II类分子。这些将是有用的 在定位细胞毒性T细胞(CTL)和辅助T细胞(HTL)反应和 猕猴的表位。申请者还将有一个小组 表达恒河猴MHC I、II类等位基因的转基因细胞 一组从已知MHC类型的动物中提取的DNA样本。这些遗嘱 在几个不同的实验室标准化MHC分型方面是有用的。这个 这份提案中的负责人在分析这两个方面都有丰富的经验 恒河猴和人类MHC。沃特金斯博士一直在研究非人灵长类(NHP) MHC I和II类等位基因，也是ASHI- 威斯康星大学医院和诊所的认证临床实验室主任 打字实验室。Bontrop博士参与了这两个人类的MHC分型工作 和猕猴在过去的15年里，是荷兰的董事 灵长类研究中心。PFI一直在提供试剂盒和试剂，用于 对过去15年人类体内MHC分子的分析。

DESCRIPTION (Adapted from the applicant's abstract): The increased utility of the rhesus macaque animal model in both HIV vaccine development and pathogenesis studies necessitates the development of reference MHC typing laboratories for this species. In this proposal the applicants plan to develop two reference laboratories for MHC typing of the rhesus macaque. Both of these laboratories will be directed by individuals who have had more than ten years of experience in analyzing the MHC of the rhesus macaque. One of these laboratories will be at the WRPRC under the direction of Dr. David Watkins and the other will be located at the Biomedical Primate Research Center (the Netherlands) under the direction of Dr. Ronald Bontrop. The applicants plan to offer services to both the North American and European scientific community for MHC typing of rhesus macaques. Initially, this will include PCR-SSP tests for alleles encoding MHC class I molecules that bind peptides derived from SIV and SHIV. The applicants will also offer PCR-SSP typing for MHC class II alleles that similarly encode MHC class II molecules that bind peptides from SIV or SHIV. The investigators propose to develop additional molecular techniques for analysis of the macaque MHC class I and class II alleles. These will include reference strand conformational analysis (RSCA) and direct sequencing of MHC class II alleles in the rhesus macaque. The investigators plan to utilize the resources of a company (PFI) that develops and markets these technologies for the analysis of the human MHC alleles. Thus, the applicants have proposed a subcontract to PFI to provide PCR-SSP plates and to develop RSCA for the rhesus macaque. Additionally, the applicants will offer training for individuals that wish to set up MHC typing in their own laboratories. Finally, the applicants will have a panel of well- characterized, MHC-defined cell lines that will be invaluable for the analysis of the MHC in the rhesus macaque. These will include a panel of rhesus macaque B-lymphoblastoid cell lines (BLCL) for which the investigators know all of the expressed MHC class I and class II molecules. These will be useful in mapping both cytotoxic T cell (CTL) and helper T cell (HTL) responses and epitopes in the macaque. The applicants will also have a panel of transfectants expressing rhesus macaque MHC class I and II alleles as well as a panel of extracted DNA samples from animals of known MHC type. These will be useful in standardizing MHC typing in several different laboratories. The principals in this proposal have extensive experience in the analysis of both rhesus and human MHC. Dr. Watkins has been studying nonhuman primate (NHP) MHC class I and II alleles for the last 15 years, and is also an ASHI- certified clinical laboratory Director at the UW Hospital and Clinics HLA Typing Laboratory. Dr. Bontrop has been involved in MHC typing in both humans and rhesus macaques for the last 15 years, and is Director of the Dutch Primate Research Center. PFI has been supplying kits and reagents for the analysis of MHC molecules in humans for the last 15 years.