High-throughput identification of common CD8+ T cell responses to SIV and M. tuberculosis in rhesus macaques

高通量鉴定恒河猴中常见 CD8 T 细胞对 SIV 和结核分枝杆菌的反应

基本信息

批准号：
10090668
负责人：
David H. O'Connor
金额：
$ 68.22万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10090668
关键词：
AIDS-Related Opportunistic Infections AIDS/HIV problem Address Algorithms Alleles Amino Acid Substitution Area Binding Biological Assay CD8-Positive T-Lymphocytes Cause of Death Communicable Diseases Communities Core Facility Cryopreservation Databases Deposition Development Ebola virus Epitopes Genbank Grant HIV HIV/TB Human Immune Influenza Institutes Interferon Type II Intervention Investments Ligands Macaca Macaca mulatta Major Histocompatibility Complex Measures Methods Modality Monkeys Mycobacterium tuberculosis National Institute of Allergy and Infectious Disease National Institute of Drug Abuse National Institute of Mental Health National Institute of Neurological Disorders and Stroke Pathogenesis Peptides Peripheral Blood Mononuclear Cell Price Proteome Reagent Research Research Personnel Research Priority Resources SIV Stains T cell response T-Lymphocyte Epitopes Technology Testing Time Tuberculosis United States National Institutes of Health Viral Genome Virus Work Zika Virus co-infection data submission enzyme linked immunospot assay high reward high risk improved in silico innovation new technology novel pathogen programs response screening

项目摘要

Project Summary/Abstract Research towards a cure for human immunodeficiency virus (HIV) and mitigation of the impact of HIV/ tuberculosis co-infections are NIH priorities. Both research areas rely on studies of macaque monkeys experimentally infected with simian immunodeficiency virus (SIV). Interventions to elicit effective CD8+ T cell responses to SIV and Mycobacterium tuberculosis (MTb) in macaques are under active develop- ment. These studies are limited by the dearth of SIV- and MTb-specific CD8+ T cell responses restricted by common macaque major histocompatibility complex (MHC) class I alleles. We began addressing this problem in the previous grant period and identified dozens of novel CD8+ T cell responses in SIV and MTb, as well as in Zika virus, ebolavirus, and influenza. We also realized that conventional T cell epitope discovery and characterization is unwieldy, slow, and cumbersome. Consequently, we demonstrated that MHC class I binding to millions of peptides can be measured si- multaneously, providing a transformative and extremely rapid way to define CD8+ T cell epitopes. The purpose of this competitive revision is to use this new technology to define CD8+ T cell epitopes in SIV and MTb restricted by 16 common macaque MHC class I alleles. Specifically, we will: Aim 1: Identify SIV and MTb CD8+ T cell responses restricted by 16 common macaque MHC class I molecules. We will assess peptide binding of each 8-, 9-, 10, and 11-mer peptide in the pro- teomes of every SIV and SHIV genome currently in Genbank and the MTb Erdman strain using an ul- tradense peptide array. MHC:peptide tetramers will be produced for responses that are experimentally validated. Aim 2: Define peptide binding motifs for the 16 common MHC class I molecules by determining the impact of every amino acid substitution at each residue in CD8+ T cell epitopes on peptide binding. The peptide binding motifs can be used to improve algorithms for in silico prediction of MHC:peptide binding. Research resources from this study will be made available to the research community through real-time sharing of peptide array data, deposition of experimentally validated CD8+ T cell epitopes in the Immune Epitope Database, and distribution of MHC:peptide tetramers through the NIH Tetramer Core Facility. Furthermore, the definition of CD8+ T cell epitopes using ultradense peptide arrays is generalizable and could revolutionize the identification of pathogen-specific epitopes in all species, including humans.

项目概要/摘要研究治愈人类免疫缺陷病毒 (HIV) 并减轻 HIV/病毒的影响结核病合并感染是 NIH 的优先事项。这两个研究领域都依赖于猕猴的研究实验性感染猿猴免疫缺陷病毒（SIV）。诱导有效 CD8+ 的干预措施猕猴中 T 细胞对 SIV 和结核分枝杆菌 (MTb) 的反应正在积极开发中。 ment。这些研究因缺乏 SIV 和 MTb 特异性 CD8+ T 细胞反应而受到限制由普通猕猴主要组织相容性复合体 (MHC) I 类等位基因组成。我们在上一个资助期开始解决这个问题，并确定了数十种新颖的 CD8+ SIV 和 MTb 以及寨卡病毒、埃博拉病毒和流感中的 T 细胞反应。我们也意识到传统的 T 细胞表位发现和表征笨拙、缓慢且繁琐。因此，我们证明了 MHC I 类与数百万个肽的结合可以通过 si- 进行测量。同时，提供了一种革命性且极其快速的方法来定义 CD8+ T 细胞表位。此次竞争性修订的目的是利用这项新技术来定义 CD8+ T 细胞表位 SIV 和 MTb 受 16 个常见猕猴 MHC I 类等位基因限制。具体来说，我们将：目标 1：识别受 16 种常见猕猴 MHC 限制的 SIV 和 MTb CD8+ T 细胞反应 I 类分子。我们将评估原蛋白中每个 8、9、10 和 11 聚体肽的肽结合。目前 Genbank 和 MTb Erdman 菌株中每个 SIV 和 SHIV 基因组的组组均使用 ul- 贸易肽阵列。 MHC：将产生肽四聚体用于实验反应已验证。目标 2：通过确定 16 种常见 MHC I 类分子的肽结合基序 CD8+ T 细胞表位中每个残基的每个氨基酸取代对肽的影响绑定。肽结合基序可用于改进计算机预测的算法 MHC：肽结合。这项研究的研究资源将通过实时方式提供给研究界共享肽阵列数据，在免疫系统中沉积经过实验验证的 CD8+ T 细胞表位表位数据库，以及 MHC：肽四聚体通过 NIH 四聚体核心设施的分布。此外，使用超密肽阵列的 CD8+ T 细胞表位的定义是可推广的并且可能会彻底改变包括人类在内的所有物种中病原体特异性表位的识别。