The Immunogenetics of Macaques Used in Biodefense Research

描述（由申请人提供）：恒河猴（Macaca mulatta）和食蟹猴（Macaca fascicularis）被广泛用作人类疾病发病机制的模型，并用于评估针对微生物病原体的疫苗和治疗方法，包括新出现的疾病和潜在的生物恐怖病原体。解释这些研究结果的能力受到准确定量其细胞免疫反应的有限能力的阻碍。本研究的重点是开发评估中国恒河猴和食蟹猴细胞介导免疫的技术和工具。我们的策略依赖于鉴定常见的主要组织相容性（MHC） I类等位基因，确定其肽结合基序，开发生物信息学工具来预测MHC：肽相互作用，并通过鉴定拉沙病毒、马尔堡病毒和流感衍生的表位来验证我们的方法。在Specific Aim 1中，我们将分别从食蟹猴和中国恒河猴中定义五种常见MHC I类分子的肽结合基序。可用于实验研究的动物队列中常见的A和B位点MHC I类等位基因频率数据将由合作者提供，并由内部生成数据。对研究的五个最常表达的MHC等位基因中的每一个都将产生稳定的转染物，从而允许可溶性MHC的生产和纯化。然后，我们将建立定量肽：MHC肽结合测定，并确定等位基因特异性MHC结合基序。这种定量MHC：肽结合数据将用于算法开发。在Specific Aim 2中，我们将开发并使MHC：肽预测算法可供科学界使用。通过Specific Aim 1获得的大量结合数据将用于生成预测多肽MHC结合能力的算法。这些工具将允许从任何感兴趣的A-C类病原体中快速预测潜在的中国起源和食蟹猴表位。所有结合数据和算法将提交给免疫表位数据库，从而使这些工具免费提供给科学界。在具体目标3中，我们将在拉沙、马尔堡和流感病毒模型系统中实验验证我们的方法。我们将测试在特异性目标1和2中进行的研究过程中发现的来自拉沙、马尔堡和流感病毒的表位的免疫原性。我们现在扩大了我们的工作范围，不仅包括最初提交的拉沙病毒，而且由于我们的计算算法质量的改进，还包括马尔堡病毒和流感病毒。这些研究将与合作者合作进行。这些实验将验证所开发的方法和工具，并确定可用于监测这些A-C病原体系统反应的表位。

DESCRIPTION (provided by applicant): Rhesus macaques (Macaca mulatta) and Cynomolgus macaques (Macaca fascicularis), are widely used as models of disease pathogenesis in humans and for evaluating vaccines and therapeutics against microbial pathogens, including emerging diseases and potential bioterror pathogens. The ability to interpret the results of these studies is hampered by a limited capacity to accurately quantitate their cellular immune responses. This proposal is focused on developing technology and tools to evaluate cell-mediated immunity in Chinese-origin rhesus macaques and Cynomolgus macaques. Our strategy relies on the identification of common major histocompatibility (MHC) class I alleles, determining their peptide binding motifs, developing bioinformatic tools to predict MHC:peptide interactions, and validating our approach by the identification of Lassa virus, Marburg virus and influenza-derived epitopes. In Specific Aim 1, we will define peptide binding motifs for five common MHC class I molecules each from Cynomolgus macaques and Chinese rhesus macaques. Common A and B loci MHC class I allele frequency data in animal cohorts available for use in experimental research will be provided by collaborators and data generated in-house. A stable transfectant will be produced for each of the five most frequently expressed MHC allele studied, allowing the production and purification of soluble MHC. We will then establish quantitative peptide:MHC peptide binding assays and determine allele specific MHC binding motifs. This quantitative MHC:peptide binding data will be used for algorithm development. In Specific Aim 2, we will develop and make MHC:peptide prediction algorithms available to the scientific community. The large amount of binding data obtained as a result of Specific Aim 1 will be utilized to generate algorithms to predict the MHC binding capacity of peptides. These tools will permit rapid predictions of potential Chinese-origin and Cynomolgus macaque epitopes from any Category A-C pathogen of interest. All binding data and algorithms will be submitted to the Immune Epitope Database, thereby making these tools freely available to the scientific community. In Specific Aim 3, we will experimentally validate our approach in the Lassa, Marburg and influenza virus model systems. We will test the immunogenicity of epitopes derived from Lassa, Marburg and influenza viruses identified in the course of the studies performed in Specific Aim 1 and 2. We now extended our efforts to encompass not only Lassa virus, as proposed in the original submission, but also Marburg and influenza viruses due to improvements in the quality of our computational algorithms. These studies will be performed in collaboration with collaborators. These experiments will validate the approach and tools developed and also identify epitopes that can be utilized to monitor responses in these A-C pathogen systems. Project Narrative: Rhesus macaques and Cynomolgus macaques are widely used as models of disease pathogenesis in humans and for evaluating vaccines and therapeutics against microbial pathogens, including emerging diseases and potential bioterror pathogens. The ability to interpret the results of these studies is hampered by a limited capacity to accurately quantitate their cellular immune responses. This proposal is focused on developing technology and tools to evaluate cell-mediated immunity in these animal models.