Mechanisms of acceptance of mouse renal allografts

小鼠同种异体肾移植物的接受机制

基本信息

批准号：
7848042
负责人：
Robert B Colvin
金额：
$ 44.19万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-22 至 2010-05-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7848042
关键词：
Address Allografting Antibodies Attention Behavior Benign Biopsy Blood Flow Cytometry Cell Communication Cells Clinical Computer Analysis Dendritic Cells Diphtheria Toxin Endothelial Cells Eragrostis Event Generations Heart Human Image Imaging Techniques Imaging technology Immune response Immunohistochemistry Infiltration Integrins Interleukin-6 Isogenic transplantation Kidney Kidney Transplantation Laboratories Lead Life Mediator of activation protein Monitor Motion Mouse Strains Mus Neonatal Organ Organ Transplantation Outcome Pathogenesis Pathologic Pathologic Processes Pathology Pharmaceutical Preparations Phenotype Physiology Process Production Property Proteins Randomized Renal tubule structure Role Saline Site Skin Transplantation Skin graft Slide Spleen System T-Lymphocyte Testing Therapeutic Time Tissues Transgenic Mice Transplantation Transplantation Tolerance Tryptophan 2,3 Dioxygenase Tubular formation Video Recording base cell behavior cell motility cell type diphtheria toxin receptor enzyme linked immunospot assay graft function heart allograft in vivo insight kidney allograft methyl tryptophan morphometry novel promoter research study response

项目摘要

This proposal addresses the great need for definitive evidence to test the prevalent dogma that Foxp3+/Tregulatory cells are essential for the acceptance of physiologically relevant MHC incompatible organ grafts. MHC-mismatched mouse renal allografts are often accepted without specific therapy in certain strain combinations, and can promote acceptance of other tissues from the same donor. Heart allografts do not have this property, suggesting an organ specific feature of the kidney. Despite demonstration of this phenomenon over 30 years ago, the mechanisms by which kidneys promote tolerance to other tissues have not been established. This project will test the role of Foxp3+ T regulatory cells and indoleamine 2,3-dioxygenase (IDO) in the pathogenesis of acceptance of mouse renal allografts, using full MHC mismatched combinations that are known to accept 33-80% of the grafts. Particular attention will be paid to interactions of Foxp3 cells with renal tubules, since Foxp3 cells concentrate in this site. Foxp3+ cells will be deleted systemically and in the graft with using the diphtheria toxin receptor (DTR) transgenic mouse with DTR expressed from the foxp3 promoter in conjunction with GFP. Retransplant experiments will test whether intragraft T cells are sufficient to reject the graft once intragraft Foxp3 cells are removed. Interaction of Foxp3+ cells with other T cells, dendritic cells and tubular cells in the renal grafts will be assessed with a real time confocal, multiphoton endomicroscope developed in our laboratory using transgenic mice expressing GFP or other fluorescent marker proteins in these cells. This system permits repeated observations of the same graft over time, live video recording, delineation of three different cell types and computer analysis of cell motility and interactions. Biopsies of renal allografts will be analyzed for cellular content (Foxp3+, Tbet+, IDO expression) in addition to standard Banff scoring as done in the human to determine which features correlate with later graft acceptance, using whole slide imaging technology and morphometric immunohistochemistry. The role of intragraft TGFβ activation, IL-6 and IDO production and CD103 expression in the generation of Treg will be assessed. Functional studies of infiltrating cells will be compared with those in the spleen for ELISPOT direct and indirect reactivity to the donor and inhibition by Foxp3+ cells.

该提案解决了对确凿证据的巨大需求，以检验普遍存在的观点 Foxp3+/Tregulatory cells 对于生理学接受至关重要相关MHC不相容的器官移植物。 MHC 不匹配的小鼠同种异体肾移植物在某些菌株组合中，通常无需特定治疗即可接受，并且可以促进接受来自同一捐赠者的其他组织。同种异体心脏没有这个性质，表明肾脏的器官特定特征。尽管示范 30多年前的这种现象，肾脏促进的机制对其他组织的耐受性尚未确定。该项目将测试角色 Foxp3+ T 调节细胞和吲哚胺 2,3-双加氧酶 (IDO) 在发病机制中的作用接受小鼠肾同种异体移植物，使用完全 MHC 不匹配的组合已知接受 33-80% 的移植物。将特别关注以下方面的互动： Foxp3 细胞与肾小管相连，因为 Foxp3 细胞集中在这个部位。 Foxp3+细胞使用白喉毒素受体将被全身删除并在移植物中删除 (DTR) 转基因小鼠，其DTR由foxp3启动子联合表达与 GFP。再移植实验将测试移植物内 T 细胞是否足以一旦移除移植物内的 Foxp3 细胞，就会排斥移植物。 Foxp3+细胞与肾移植物中的其他 T 细胞、树突状细胞和肾小管细胞将通过我们实验室开发的实时共焦多光子内窥镜在这些细胞中表达 GFP 或其他荧光标记蛋白的转基因小鼠。这系统允许随着时间的推移重复观察同一移植物，实时视频记录，三种不同细胞类型的划分以及细胞运动和细胞运动的计算机分析互动。将分析肾同种异体移植物的活组织检查中的细胞含量（Foxp3+、 Tbet+、IDO 表达）以及在人类中进行的标准班夫评分使用全玻片成像确定哪些特征与后期移植物接受相关技术和形态测定免疫组织化学。移植物内 TGFβ 的作用 Treg 生成过程中的激活、IL-6 和 IDO 的产生以及 CD103 的表达将进行评估。浸润细胞的功能研究将与浸润细胞的功能研究进行比较脾脏对供体的 ELISPOT 直接和间接反应性以及 Foxp3+ 的抑制细胞。