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.

项目摘要/摘要治愈人类免疫缺陷病毒（HIV）的研究以及艾滋病毒/ 结核病共感染是NIH的优先级。这两个研究领域都依赖于猕猴的研究实验感染了猿猴免疫缺陷病毒（SIV）。引起有效CD8+的干预措施 T细胞对猕猴中SIV和结核分枝杆菌（MTB）的反应在主动发展下精神。这些研究受到SIV和MTB特异性CD8+ T细胞反应的缺乏的限制通过普通猕猴的主要组织相容性复合物（MHC）I类等位基因。我们开始在上一个赠款期间解决这个问题，并确定了数十个新颖的CD8+ SIV和MTB中的T细胞反应以及Zika病毒，埃博氏病毒和流感。我们也意识到传统的T细胞表位发现和表征是笨拙，缓慢且繁琐的。因此，我们证明了MHC I类与数百万肽的结合可以测量多种多样，提供了一种定义CD8+ T细胞表位的变革性且极快的方法。这种竞争性修订的目的是使用这项新技术定义CD8+ T细胞表位 SIV和MTB受16个常见的猕猴MHC I类等位基因限制。具体来说，我们将： AIM 1：识别SIV和MTB CD8+ T细胞响应受16个常见猕猴MHC的限制 I类分子。我们将评估促值中每种8，9-，10和11-mer肽的肽结合 GenBank和MTB Erdman菌株目前每个SIV和SHIV基因组的TEOMES都使用UL- Tradense肽阵列。 MHC：将生产肽四聚体的反应经过验证。 AIM 2：通过确定16个常见MHC I类分子的肽结合基序每种氨基酸取代CD8+ T细胞表位对肽中的每个残基的影响结合。肽结合基序可用于改善用于硅预测的算法 MHC：肽结合。这项研究的研究资源将通过实时提供给研究社区共享肽阵列数据，在免疫中实验验证的CD8+ T细胞表位沉积 EpiTope数据库和MHC的分布：通过NIH四聚体核心设施的肽四聚体。此外，使用超知肽阵列的CD8+ T细胞表位定义是可推广的，并且可以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鉴定。