Tracking anti-fungal CD4+ T cells in vivo

体内追踪抗真菌 CD4 T 细胞

基本信息

批准号：
8610881
负责人：
BRUCE Steven KLEIN
金额：
$ 22.58万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8610881
关键词：
Address Adoptive Transfer American Antifungal Agents Antigens Attenuated Live Virus Vaccine Biology Blastomyces Blastomyces dermatitidis Blastomycosis Build-it C57BL/6 Mouse CD4 Positive T Lymphocytes CD8B1 gene Calnexin Cause of Death Cells Cessation of life Clinical Coccidioides Coccidioides posadasii Coccidioidomycosis Data Development Disease Engineering Environment Epitopes Evolution Exposure to Fungal Antigens Glean Goals Histoplasma Histoplasmosis Immunity Immunodominant Epitopes Immunologic Memory Immunology Individual Infection Infection prevention Interleukin-17 Knowledge Life Longevity Lung MHC Class II Genes Measures Mediating Medicine Memory Microbe Monitor Mus Mycoses Nature Paracoccidioidomycosis Peptide/MHC Complex Peptides Persons Phenotype Physiological Population Prevention strategy Public Health Reagent Reporting Resistance Soil Sporotrichosis Stem cells System T-Lymphocyte T-Lymphocyte Epitopes Testing The science of Mycology Therapeutic Transgenic Mice Transgenic Organisms Tuberculosis Vaccinated Vaccination Vaccine Design Vaccines Work base cytokine disorder prevention fungus in vivo insight interest memory CD4 T lymphocyte mortality novel pathogen prevent public health relevance

项目摘要

DESCRIPTION (provided by applicant): We propose an exploratory project to create peptide MHC II tetramers that will advance insight into the immunity to endemic mycoses and the biology of long-lived memory among IL-17 producing CD4+ T cells. Ag- specific T cells that enter the pool of long-lived memory cells represent the cellular basis of immunological memory, which is the underpinning of vaccine- and protective-immunity. An understanding of T cells that persist in this pool is needed for the rationale design of vaccines. There are differing views about the longevity of T helper 17 cells. Amid this debate, there's a knowledge gap about CD4+ T cells that enter the memory pool after fungal vaccination, and whether the cells retain the ability to produce IL-17. This knowledge gap is a barrier to disease prevention strategies since Th17 cells promote vaccine immunity to fungi. We hypothesize that Ag-specific CD4+ T cells that protect against dimorphic fungi - Blastomyces, Histoplasma, and Coccidioides - enter the memory pool and retain the ability long-term to produce IL-17 on re-exposure to the pathogen. We'll test our hypothesis by using a novel TCR transgenic mouse to decipher the identity of a dominant, protective Ag conserved in dimorphic fungi, and introduce the antigen's T cell epitope into peptide-MHC (pMHC) tetramers to track the evolution, persistence, phenotype and function of endogenous Ag-specific CD4+ T cells in vivo under physiological conditions after fungal vaccination and re-exposure in the lung. Our aims are to: 1) Identify the shared protective fungal Ag and epitope and create and validate peptide MHC tetramers. We will identify, clone and express the fungal Ag recognized by vaccine induced CD4+ T cells that mediate protective immunity to endemic mycoses. Our preliminary data point strongly toward calnexin. We'll deduce the peptide epitope of the Ag recognized by the class II MHC of BL/6 mice and synthesize and validate pMHC tetramers to let us monitor and track endogenous epitope specific CD4+ T cells under physiological conditions. 2) Define the persistence and phenotype of vaccine-induced anti-fungal T17 cells using pMHC tetramers. We will use pMHC tetramers to define the size of the endogenous pool of epitope-specific CD4+ T cells in na¿ve BL/6 mice. In vaccinated mice, we will use pMHC tetramers to track the long-term fate, persistence and phenotype of memory CD4+ T cells that mediate resistance to the mycoses, with an emphasis on clarifying the extent to which Th17 cells persist after vaccination to blastomycosis, histoplasmosis, and coccidioidomycosis. Our work will yield new insight into the phenotype and function of CD4+ T cells that confer protective immunity to the endemic mycoses, and especially the persistence of IL-17 producing cells over an extended period. The results will be broadly significant and impactful for those working in Mycology & Immunology.

描述（由适用提供）：我们提出了一个探索性项目，以创建肽MHC II四聚体，以提高对内部分子真菌的免疫学的深入了解，以及在产生CD4+ T细胞的IL-17中长寿记忆的生物学。进入长寿记忆细胞池的Ag特异性T细胞代表了免疫记忆的细胞基础，这是疫苗和受保护的免疫力的基础。对于疫苗的理由设计，需要对持续存在的T细胞的理解。关于T辅助17个细胞的寿命有不同的看法。在这场辩论中，存在有关CD4+ T细胞的知识差距，该细胞在真菌疫苗后进入记忆池，以及细胞是否保留生产IL-17的能力。由于Th17细胞促进了对真菌的疫苗免疫，因此这种知识差距是预防疾病策略的障碍。我们假设可以预防二态真菌 - 胚泡，组织肿瘤和球孢菌素的Ag特异性CD4+ T细胞进入记忆库，并保留长期生产IL-17在重新暴露于病原体时产生IL-17的能力。我们将通过使用新型的TCR转基因小鼠来检验我们的假设，以解释在双态真菌中保守的主导，受保护的Ag的身份，并将抗原的T细胞表位引入肽-MHC（PMHC）Tetramers中，以跟踪进化，持久性，持久性，表现型和内源性Ag-demogitif CD4+ TING CD4+ TIN GUIC CD4+ TIVO CDOC中的功能。在肺部。我们的目标是：1）确定共享的受保护的真菌Ag和表位，并创建和验证肽MHC四聚体。我们将识别，克隆并表达疫苗诱导的CD4+ T细胞识别的真菌AG，这些真菌介导了受保护对内在真菌剂的免疫力。我们的初步数据强烈指向钙网蛋白。我们将推断出BL/6小鼠II类MHC识别的Ag的肽发作，并合成并验证PMHC四聚体，使我们在物理条件下监测和跟踪内源性表位特异性CD4+ T细胞。 2）使用PMHC四聚体定义疫苗诱导的抗真菌T17细胞的持久性和表型。我们将使用PMHC四聚体来定义Na�VEBL/6小鼠中表位特异性CD4+ T细胞的内源性池的大小。在接种疫苗的小鼠中，我们将使用PMHC四聚体来跟踪记忆CD4+ T细胞的长期命运，持久性和表型，这些命中介导了对真菌的抗性，重点是阐明TH17细胞在疫苗接种后对疫苗接种后对疫苗的疫苗接种的程度，对大口胞菌病，组织质质膜病和cocciciyilesymycosis进行了疫苗接种。我们的工作将对CD4+ T细胞的表型和功能产生新的见解，这些表型和功能会导致对内部分子真菌的保护性免疫，尤其是在很长时间内IL-17产生细胞的持久性。结果对于在真菌学和免疫学从事的人来说将具有广泛的意义和影响力。