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

项目概要/摘要 此应用程序是一个指导病人为导向的研究职业发展奖，题为“影响 组织驻留记忆T细胞对肺移植后慢性排斥反应的影响”，由Mark博士提交 斯奈德，医学和免疫学助理教授在肺，过敏，和 匹兹堡大学重症监护医学，Starzl移植研究所成员。的 本申请书中列出的短期目标旨在帮助申请人实现其长期目标 成为人类肺部免疫学研究领域的独立研究者和领导者。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基础，（2） 扩展有效执行转化免疫学所需的技术和分析工具 研究，和（3）发展领导技能，需要运行一个生产性的人类免疫学实验室。 这项研究的主要目的是研究同种异体移植组织驻留记忆T 细胞（TRM）的持久性和一代的风险发展闭塞性细支气管炎综合征，主要 慢性肺移植物功能障碍（CLAD）的表型表现。CLAD影响高达50%的肺 移植后5年内存活，并与早期死亡率和大量 发病率CLAD的机制仍不明确，但被认为是T细胞介导的过程。 先前的急性细胞排斥反应（ACR）和感染，这两种T细胞介导的过程，与 CLAD的最终发展。申请人之前的工作，最近发表在《科学免疫学》上， 表明肺同种异体移植物中供体TRM持续性和受体TRM产生与以下因素相关： ACR和感染。此外，未公布的初步数据显示， CLAD在接受TRM的快速同种异体移植人群中的作用。根据这项建议，申请人 在此基础上，本文提出了以下具体目标：（1）确定接收方TRM之间的关系 肺同种异体移植物中的世代和早期CLAD的风险（移植后2年内），（2）确定 同种异体反应潜力和病原体特异性，以及（3）确定关系 T细胞受体（TCR）克隆多样性和基因表达之间的受体移植TRM和CLAD。 主要假设是受体TRM累积与早发CLAD相关， 受体TRM将由具有高同种异体反应潜力的扩增的T细胞群组成。到 为了实现这些目标，申请人将采用其先前公布的分离供体的方法， 受体TRM从肺移植受体的支气管肺泡灌洗纵向。cox比例 将进行风险模型，调整已知混杂因素，以检验暴露（早期比例 与受体TRM相比，供体TRM下降），因为它与我们早期CLAD的结果有关。混合淋巴细胞 将进行TRM反应和单细胞TCR测序以确定TRM同种异体反应性和克隆性。

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.