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.

这项建议解决了对确凿证据的巨大需求，以测试流行的 信条认为Foxp3+/T调节细胞是接受生理上的 相关MHC血型不合的器官移植。MHC不匹配的小鼠肾移植 在某些菌株组合中通常在没有特定治疗的情况下被接受，并可以促进 接受来自同一捐赠者的其他组织。同种异体心脏移植没有这个 财产，暗示肾脏的器官特有的特征。尽管展示了 这一现象在30多年前，肾脏促进的机制 对其他组织的耐受性尚未建立。该项目将测试 Foxp3+T调节细胞和吲哚胺2，3-双加氧酶在慢性粒细胞白血病发病机制中的作用 接受小鼠同种异体肾移植，使用完全MHC不匹配的组合 已知接受33%-80%的移植物。我们将特别关注 有肾小管的Foxp3细胞，因为Foxp3细胞集中在这个部位。Foxp3+细胞 将在使用白喉毒素受体的移植物中被系统删除 联合表达Foxp3启动子的(DTR)转基因小鼠 使用绿色荧光蛋白。再次移植实验将测试移植内的T细胞是否足以 一旦移植物内的Foxp3细胞被移除，就拒绝移植物。Foxp3+细胞与Foxp3+细胞相互作用 移植肾中的其他T细胞、树突状细胞和肾小管细胞将通过 本实验室研制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相同的移植物，实况视频记录， 三种不同细胞类型的划分和细胞运动和细胞运动的计算机分析 互动。将分析移植肾的活检组织的细胞含量(Foxp3+， Tbet+，IDO表达)除了在人类To中所做的标准Banff评分之外 使用全幻灯片成像，确定哪些特征与移植后的接受性相关 技术和形态计量免疫组织化学。移植物内转化生长因子的作用 Treg Will的激活、IL-6和IDO的产生及CD103的表达 被评估。浸润性细胞的功能研究将与 ELISPOT脾对供体的直接和间接反应及Foxp3+的抑制作用 细胞。