Mechanisms of acceptance of mouse renal allografts

小鼠同种异体肾移植物的接受机制

• 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
• 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

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.

该提案涉及确定证据的巨大需求以测试普遍存在 foxp3+/tregulatory细胞的教条对于接受生理学至关重要 相关的MHC不兼容器官移植。 MHC不匹配的小鼠肾同种异体移植物是 在某些应变组合中通常不接受特定治疗的接受，并可以促进 接受来自同一供体的其他组织。心脏同种异体没有这个 属性，暗示了肾脏的特定器官特征。尽管展示了 30年前，这种现象是肾脏促进的机制 尚未建立对其他组织的耐受性。该项目将测试 Foxp3+ T调节细胞和吲哚胺2,3-二氧酶（IDO）在发病机理中 使用完全MHC不匹配的组合的小鼠肾同种异体移植物接受 已知接受33-80％的移植物。特别关注的是 由于Foxp3细胞集中在该部位，因此带有肾小管的Foxp3细胞。 FOXP3+细胞 使用白喉毒素受体将系统地删除和在移植物中删除 （DTR）用DTR从FOXP3启动子结合使用的DTR的（DTR） 与GFP。后期植物实验将测试Intraft T细胞是否足以 一旦移除了foxp3细胞，拒绝移植物。 Foxp3+细胞与 其他T细胞，肾移植物中的树突状细胞和管状细胞将通过A进行评估 实时共聚焦，多光子内分球体在我们的实验室开发的使用 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时间对同一移植物的重复观察，实时视频录制， 描述了三种不同的细胞类型和计算机分析细胞运动和计算机分析 互动。将分析肾脏同种异体移植活检的细胞含量（Foxp3+， 除了在人类中完成的标准班夫评分外，tbet+，ido表达式） 使用整个幻灯片成像确定哪些功能与后来的移植物接受相关 技术和形态计量学免疫组织化学。 IntraftTGFβ的作用 激活，IL-6和IDO的产生以及Treg产生的CD103表达将 被评估。将浸润细胞的功能研究与 ELISPOT直接和间接反应性的脾 细胞。