MHC TYPING OF MACAQUES USED IN AIDS RESEARCH

描述（改编自申请人的摘要）： 恒河猴动物模型在HIV疫苗开发和 发病机制的研究需要参考MHC分型的发展 这个物种的实验室。 在本提案中，申请人计划 建立两个恒河猴MHC分型参考实验室。 两 这些实验室将由个人谁拥有超过 有十年分析恒河猴MHC的经验 之一 这些实验室将在大卫博士的指导下在WRPRC工作 沃特金斯和另一个将位于生物医学灵长类动物研究所 中心（荷兰）在罗纳德邦特罗普博士的指导下。 的 申请人计划向北美和欧洲提供服务， 猕猴MHC分型的科学界。 最初，这将 包括编码MHC I类分子等位基因的PCR-SSP测试，所述MHC I类分子结合 来源于SIV和SHIV的肽。 申请人还将提供PCR-SSP MHC-II类等位基因的分型，其类似地编码MHC-II类分子 结合SIV或SHIV的肽。 研究人员建议开发 用于分析猕猴MHC 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）和恒河猴中MHC II类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，申请人向PFI提出了一项建议， PCR-SSP平板和开发恒河猴的RSCA。 另夕h 申请人将为希望建立MHC分型的个人提供培训 在他们自己的实验室里。 最后，申请人将有一个良好的小组- 特征性的，MHC定义的细胞系，这将是非常宝贵的分析 在恒河猴体内的主要组织相容性复合体 其中包括一组恒河猴 猕猴B淋巴母细胞样细胞系（BLCL），研究人员知道， 所有表达的MHC I类和II类分子。 这些会有用的 在绘制细胞毒性T细胞（CTL）和辅助性T细胞（HTL）应答中， 猕猴体内的抗原决定簇 申请人还将有一个小组， 表达恒河猴MHC I类和II类等位基因以及 一组从已知MHC类型的动物中提取的DNA样品。 这些将 在几个不同的实验室标准化MHC分型是有用的。 的 本提案的主要负责人在分析这两方面都有丰富的经验， 恒河猴和人MHC。 沃特金斯博士一直在研究非人类灵长类动物（NHP） MHC I类和II类等位基因，也是一种ASHI- 华盛顿大学医院和诊所HLA认证临床实验室主任 打字实验室。 Bontrop博士参与了两种人类的MHC分型 在过去的15年里，他是荷兰 灵长类研究中心。 PFI一直在为 在过去的15年里，人类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.