Impact of tissue resident memory T cells on chronic rejection after lung transplantation

项目摘要/摘要 本申请表是关于以患者为导向的研究事业发展奖的申请表，标题为 肺移植后慢性排斥反应中的组织驻留记忆T细胞“，由Mark博士提交 斯奈德，肺，过敏和免疫学分部的医学和免疫学助理教授 匹兹堡大学重症监护医学博士，斯塔兹尔移植研究所成员。这个 本意见书中概述的短期目标旨在帮助申请者实现其长期目标。 成为人类肺部免疫学研究领域的独立研究员和领导者。这些 短期目标包括(1)推进与适应性免疫和先天免疫相关的知识库，(2) 有效执行翻译免疫学所需的技术和分析工具的扩展 研究，以及(3)开发领导技能，以运行一个富有成效的人类免疫学实验室。 本研究方案的中心目标是研究同种异体移植组织驻留记忆T的影响 细胞(TRM)持续性与代发闭塞性毛细支气管炎综合征的风险 慢性同种异体肺移植功能障碍的表型表现。包衣对肺部的影响高达50% 移植后5年存活，并与早期死亡率和 发病率。CLAD的机制尚不清楚，但被认为是T细胞介导的过程。 前驱急性细胞排斥反应(ACR)和感染，两者都是T细胞介导的过程，与 衣着的终极发展。申请者之前的工作最近发表在《科学免疫学》上， 表明供体TRM的持久性和受者在肺移植中TRM的生成与 急性呼吸窘迫综合征和感染。此外，未公布的初步数据显示出早期发展的趋势。 在那些接受TRM的同种异体移植患者中，CLAD的存在。有了这项建议，申请人 以以下具体目标对先前的工作进行扩展：(1)定义接收方TRM之间的关系 同种异体肺移植的生成和早期包膜的风险(移植后2年内)，(2)决定 受体来源的同种异体移植物的同种异体反应潜能和病原体特异性，以及(3)确定两者之间的关系 T细胞受体(TCR)克隆多样性与受体基因表达的关系 主要假设是受者TRM积聚将与早发性包裹体相关，并且 受体TRM将由具有高同种异体反应潜能的T细胞扩大群体组成。至 为了达到这些目的，申请人将使用他以前发表的隔离捐赠者和 受体TRM来自肺移植受者纵向的支气管肺泡灌洗。考克斯比例 危险模型，对已知混杂因素进行调整，以测试暴露(早期比例 供者的TRM与受者的TRM相比下降)，因为这与我们早期覆盖的结果有关。混合淋巴细胞 将进行反应和单细胞TCR测序，以确定TRM的同种反应性和克隆性。

Project Summary / Abstract This application is for a Mentored Patient-Oriented Research Career Development Award entitled, “The impact of tissue resident memory T cells on chronic rejection after lung transplantation”, submitted by Dr. Mark Snyder, an Assistant Professor of Medicine and Immunology within the Division of Pulmonary, Allergy, and Critical Care Medicine at the University of Pittsburgh, and member of the Starzl Transplantation Institute. The short-term goals outlined in this submission are designed to help the applicant achieve his long-term objective of becoming an independent investigator and leader in the field of human lung immunology research. These short-term goals include (1) advancing knowledge base related to both adaptive and innate immunity, (2) expansion of both technical and analytic tools required to effectively perform translational immunology research, and (3) development of leadership skills required to run a productive human immunology laboratory. The central objective of this research proposal is to investigate the impact of allograft tissue resident memory T cell (TRM) persistence and generation on the risk of developing bronchiolitis obliterans syndrome, the major phenotypic presentation of chronic lung allograft dysfunction (CLAD). CLAD affects up to 50% of lung transplant survivors by 5 years after transplantation and is associated with early mortality and substantial morbidity. The mechanism of CLAD remains undefined but is believed to be a T cell-mediated process. Antecedent acute cellular rejection (ACR) and infection, both T cell mediated processes, are associated with the ultimate development of CLAD. The applicant’s prior work, recently published in Science Immunology, shows that donor TRM persistence and recipient TRM generation within the lung allograft are associated with both ACR and infections. Furthermore, unpublished preliminary data show a trend towards early development of CLAD in those patients with rapid allograft population of recipient TRM. With this proposal, the applicant expands on this prior work with the following specific aims (1) Define the relationship between recipient TRM generation in the lung allograft and the risk of early CLAD (within 2 years of transplantation), (2) Determine the alloreactive potential and pathogen specificity of recipient-derived allograft TRM, and (3) Identify the relationship between T cell receptor (TCR) clonal diversity and gene expression among recipient allograft TRM and CLAD. The primary hypothesize is that recipient TRM accumulation will be associated with early-onset CLAD and that recipient TRM will be composed of an expanded population of T cells with high allo-reactive potential. To accomplish these aims, the applicant will employ his previously published method of isolating donor and recipient TRM from the bronchoalveolar lavage of lung transplant recipients longitudinally. Cox-proportional hazard model, adjusting for known confounders will be performed to test the exposure (early proportional decline in donor TRM compared to recipient TRM) as it relates to our outcome of early CLAD. Mixed lymphocyte reactions and single-cell TCR sequencing will be performed to determine TRM alloreactivity and clonality.