P-glycoprotein Function in Allograft Rejection

简介(申请人提供)：申请人为一名肾病专科医生，患有 对移植免疫学感兴趣的人，谁有资格并满足所有 临床指导型科学家应考虑的必要条件 奖品。作为他在布里格姆妇女医院的肾脏科研究员的一部分 医院和波士顿儿童医院，申请者完成3 多年的基于实验室的研究，在此期间他获得了实践经验 在细胞和分子免疫学技术方面。他的主旨是 对P-糖蛋白的作用进行了研究，这种多药 同种异体免疫中耐药相关的ABC转运蛋白及其原理 研究结果现已发表在《免疫学杂志》上。申请人 现已完成他的肾病研究，并将被任命为工作人员 布里格姆妇女医院会员和医学讲师 哈佛医学院2001年7月1日生效。在这份建议书中，申请人 希望以他的发现为基础，即P-糖蛋白在 体外T细胞活化和APC功能。他发现了特定的P- 糖蛋白阻断抑制同种异体抗原依赖的T细胞，但不抑制丝裂原依赖的T细胞 通过正反馈循环抑制干扰素-γ和白介素12的激活， 从而阻断了启动初级同种异体免疫的关键途径 回应。这些机制也可能在体内和P-糖蛋白中起作用 可能在临床同种异体移植排斥反应中起重要作用。然而， P-糖蛋白在T细胞活化和同种异体识别中的体内作用 还没有被探索过。此外，分子信号和机制 T淋巴细胞对P-糖蛋白表达的调节作用 同种异体免疫相互作用目前尚不清楚。这项提议的目标是 目的：1)明确P-糖蛋白在直接和间接途径中的作用 同种异体识别和体内移植排斥反应的小鼠模型 2)研究P-糖蛋白在同种异体移植中的体内功能。 用小鼠过继转移系统激活抗原特异性T细胞 在这种情况下，一小部分特异性的CD4+TCR转基因T细胞可以 进行身体跟踪和表型检查；以及3)体外检查 P-糖蛋白在抗原提呈细胞功能和胆汁淤积症中的作用 T细胞和同种异体内皮细胞的定向激活是关键 对于同种异体移植排斥反应，根据初步鉴定的唯一 同种异体内皮细胞触发T细胞P-糖蛋白的能力 表情。申请者在分子生物学和细胞学方面的培训 免疫学使他发现了P-糖蛋白在脑内的一种新作用 同种异体豁免。申请人意识到，为了进一步探索他的 最近的研究发现，他需要更多的指导。被指导的临床科学家 发展奖将为申请者提供关键的机会 进一步扩大他在免疫学方面的知识，以实现 独立。导师的实验室和合作导师的实验室提供了理想的 实现本项目目标的环境，因为其强大的 移植免疫学背景，他们的合作势头和 他们模范的说教和道德课程。在完成这个项目之后 申请人应熟悉体内和体外的关键技术。 分子和细胞免疫学技术，以检测 独立和竞争性地发挥相关或新的分子靶标的作用 在移植和同种异体排斥反应中。

DESCRIPTION (provided by applicant): The applicant is a nephrologist with an interest in transplant immunology, who is eligible for and fulfills all the necessary requirements to be considered for a Mentored Clinical Scientist Award. As part of his nephrology fellowship at the Brigham and Women's Hospital and The Children's Hospital in Boston, the applicant completed 3 years of laboratory-based research during which he gained practical experience in techniques of cellular and molecular immunology. The main thrust of his research has been the study of the role of P-glycoprotein, the multidrug resistance-associated ABC transporter, in alloimmunity, and the principle findings are now being published in the Journal of Immunology. The applicant is now completing his nephrology fellowship and will be appointed as a staff member at the Brigham and Women's Hospital and as an Instructor of Medicine at Harvard Medical School effective July 1, 2001. In this proposal, the applicant wishes to build on his findings that P-glycoprotein serves critical roles in vitro in T cell activation and APC function. He has found that specific P- glycoprotein blockade inhibits alloantigen- but not mitogen-dependent T cell activation by inhibiting IFN-gamma and IL-12 through a positive feedback loop, thus blocking a critical pathway in the initiation of primary alloimmune responses. These mechanisms may also be operative in vivo and P-glycoprotein may therefore play an important role in clinical allograft rejection. However, the in vivo role of P-glycoprotein in T cell activation and allorecognition has not been explored. In addition, the molecular signals and mechanisms that regulate P-glycoprotein expression by T lymphocytes in the course of alloimmune interactions are currently unknown. The goals of this proposal are to: 1) define the role of P-glycoprotein in the direct and indirect pathways of allorecognition and in allograft rejection in vivo in murine models of allotransplantation; 2) investigate the in vivo function of P-glycoprotein in antigen-specific T cell activation using a murine adoptive transfer system where a small population of peptide-specific CD4+ TCR transgenic T cells can be physically tracked and phenotypically examined; and 3) examine in vitro the role of P-glycoprotein in antigen presenting cell function and in the bi- directional activation of T cells and allogeneic endothelium which is critical for allograft rejection, based on the preliminary identification of a unique capacity of allogeneic endothelium to trigger T cellular P-glycoprotein expression. The applicant's training in molecular biology and cellular immunology has allowed him to discover a novel role of P-glycoprotein in alloimmunity. The applicant realizes that in order to further explore his recent findings he needs additional mentoring. The Mentored Clinical Scientist Development Award will provide the critical opportunity for the applicant to further expand his knowledge in immunology necessary for achieving independence. The Mentor's and the Co-Mentor's laboratories provide an ideal environment to accomplish the aims of this project because of their strong background in transplantation immunology, their collaborative momentum and their exemplary didactic and ethical curricula. After completing this project the applicant will be familiar with the critical in vitro and in vivo techniques of molecular and cellular immunology in order to examine independently and competitively the role of related or novel molecular targets in transplantation and allograft rejection.