CD8 Responses in situations of limited T-cell repertoire

T 细胞库有限情况下的 CD8 反应

基本信息

批准号：
8376416
负责人：
Michael J Bevan
金额：
$ 41.32万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8376416
关键词：
Ablation Affinity Age Animals Antigens Automobile Driving Avidity Binding Biological Assay CD8B1 gene Cells Epitopes Failure Genetic Goals Histocompatibility Antigens Class I Immune Immune system Immunity Individual Infection Ligands Listeria monocytogenes Lymphocytic choriomeningitis virus Lymphoid Memory Mus OVA-8 Outcome Ovalbumin Pacific Northwest Peptide/MHC Complex Peptides Phenotype Recombinants Role Staining method Stains T cell response T-Lymphocyte Transgenic Organisms Variant adaptive immunity aged cytokine in vivo killings novel pathogen research study response

Ablation Affinity Age Animals Antigens Automobile Driving Avidity Binding Biological Assay CD8B1 gene Cells Epitopes Failure Genetic Goals Histocompatibility Antigens Class I Immune Immune system Immunity Individual Infection Ligands Listeria monocytogenes Lymphocytic choriomeningitis virus Lymphoid Memory Mus OVA-8 Outcome Ovalbumin Pacific Northwest Peptide/MHC Complex Peptides Phenotype Recombinants Role Staining method Stains T cell response T-Lymphocyte Transgenic Organisms Variant adaptive immunity aged cytokine in vivo killings novel pathogen research study response

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项目摘要

In this proposal, we plan to explore some of the consequences that arise when individuals with a limited or skewed CD8" T cell TCR repertoire encounter novel pathogen epitopes. Our focus is on the role of TCR affinity for its MHC/peptide ligand in driving the CDS response. We postulate that the apparent failure of immune compromised animals to mount a robust peak response may obscure a low avidity CD8+ T cell response that occurs but is missed by the usual ways of performing tetramer staining, target killing, and intracellular cytokine staining (ICS). The goal of the project is to discover whether immune-compromised animals (such as aged, or animals recovered from lymphoid ablation therapies) mount low avidity CDS responses, how these T cells are controlled, how tightly they must recognize the pathogen to contribute to immunity, and how they influence the outcome of infection. SPECIFIC AIM 1. To characterize the phenotype, function and genetic signature of TCR transgenic CD8+ T cells responding in vivo to infection with a pathogen expressing either a high or a low affinity TCR ligand. For these experiments, we have generated a number of recombinant strains of the bacterial pathogen, Listeria monocytogenes, which secrete hen Ovalbumin (Ova) containing either the wild-type peptide (SIINFEKL) recognized by the OT-1 TCR, or single residue variants (altered peptide ligands) that interact less well with this TCR while binding equally well to the relevant H2-Kb MHC class I molecule. SPECIFIC AIM 2. To assess the protection provided by T cells with a relatively low avidity for pathogen epitopes. Protection against a challenge infection with Listeria monocytogenes expressing a range of altered peptide ligands recognized by TCR transgenic cells will be assayed in these experiments. SPECIFIC AIM 3. To determine the functional avidity of T cells responding to infection in mice with limited TCR repertoires and the consequences for immune protection. Limited repertoires will be generated in a number of ways including use of old mice, thymectomized mice, in some cases primed with pathogens such as lymphocytic choriomeningitis virus (LCMV) that evoke large effector and memory CD8+ T cell responses, and mice recovering from lymphoid ablation. Infection with pathogens that are either unrelated or crossreactive with previously encountered pathogens will be used.

在此提案中，我们计划探讨当有限的人或偏斜的CD8“ T细胞TCR曲目会遇到新的病原体表位。我们的重点是TCR的作用在驱动CDS响应时，其MHC/肽配体的亲和力。我们假设明显的失败免疫受损的动物以安装强大的峰值响应可能会掩盖低自发的CD8+ T细胞发生的反应，但由于通常的方法杀死四聚体染色，杀死目标和细胞内细胞因子染色（ICS）。该项目的目的是发现免疫受损动物（例如老年人，或从淋巴消融疗法中回收的动物）持续低自发CD 响应，如何控制这些T细胞，它们必须认识到病原体的紧密程度免疫力以及它们如何影响感染结果。特定目的1。表征TCR转基因CD8+ T的表型，功能和遗传特征在体内反应的细胞以表达高亲和力TCR配体的病原体感染。对于这些实验，我们已经产生了许多重组菌株的细菌病原体，单核细胞增生李斯特菌，分泌含有野生型肽的雌雄同体（OVA）（siinfekl）由OT-1 TCR识别或单个残基变体（改变的肽配体）相互作用该TCR与相关的H2-KB MHC I类分子的结合较不太良好。特定目的2。评估病原体相对较低的T细胞提供的保护表位。防止用单核细胞增生李斯特菌感染挑战感染，表达一系列改变在这些实验中，将分析由TCR转基因细胞识别的肽配体。特定目的3。确定T细胞对小鼠感染的功能性亲和力，有限 TCR曲目和免疫保护的后果。有限的曲目将在在某些情况下，有多种方法，包括使用旧小鼠，胸腺切除小鼠，在某些情况下以病原体为底漆作为淋巴细胞绒毛膜炎病毒（LCMV），引起大型效应子和记忆CD8+ T细胞反应，和从淋巴消融中恢复的小鼠。感染无关或交叉反应的病原体将使用先前遇到的病原体。