Apoptotic Donor Leukocytes to Promote Kidney Transplant Tolerance

项目2概要 我们证明了两次移植围术期输注凋亡供体白细胞（ADL）和短暂的 用α-CD 40、雷帕霉素、sTNFR和α-IL-6 R进行免疫抑制（TIS）可诱导长期（>1年）耐受 5例未致敏、1例MHC-II DRB等位基因匹配的非人灵长类动物（NHP）中，5例接受胰岛同种异体移植。计划2 将检查ADL+TIS方案在肾移植模型中的疗效和机制，并应用高- 三维免疫分析，以指导方案的完善，主要目的是开发一种安全， 在活体供肾移植中诱导稳定耐受的有效且临床可翻译的方案。 我们假设，ADL+TIS方案，有必要进行修改，以促进成功 转化为实体器官移植环境，并根据整个U19项目的结果进行改进， 通过操作耐受性促进NHP活体供肾移植模型中移植物的长期存活。 为了检验这一假设并促进ADL+TIS的临床转化，我们提出了两个具体目标： 目的#1：确定ADL输注联合短暂性 免疫抑制在NHP肾移植模型中诱导和维持耐受作用 确定ADL+TIS方案在实现肾移植手术耐受中的有效性的研究 NHP将伴随血液、移植物、脾脏和尿液的深度免疫分析，以研究其影响 关于不同的效应物、耗尽的和调节性免疫的丰度和活化概况的方案 移植物和脾脏内不同区室中的细胞亚群及其空间组织。 目的#2：研究炎性小体抑制和IL-1b拮抗对 促进NHP中ADL输注和短暂免疫抑制诱导的肾移植耐受 这方面的研究将确定抑制炎性小体及其产物的策略的能力， 在NHP肾移植模型中，与ADL+TIS协同促进手术耐受。机械论 研究将调查炎性小体抑制和IL-1b拮抗作用如何改变ADL + TIS对 血液、尿液和脾脏中髓样和淋巴样细胞亚群的频率和活化谱， 在肾移植物中的协调免疫调节结构域中的募集和空间相互作用。 该提案的意义和创新在于ADL+TIS方案的有效性， 同种异体特异性T细胞和创造有效的免疫调节。肾移植耐受诱导的前景 根据啮齿动物肾移植和胰岛移植的耐受性记录， 在NHP和机会，合理完善的战略所创造的系统水平的免疫分析。

PROJECT 2 SUMMARY We demonstrated that two peritransplant infusions of apoptotic donor leukocytes (ADLs) and transient immunosuppression (TIS) with α-CD40, rapamycin, sTNFR, and α-IL-6R induced long-term (>1 year) tolerance to islet allografts in 5 of 5 nonsensitized, 1 MHC-II DRB allele-matched nonhuman primates (NHPs). Project 2 will examine the efficacy and mechanisms of the ADL+TIS regimen in a kidney transplant model and apply high- dimensional immune profiling to guide protocol refinements, with the PRINCIPAL OBJECTIVE of developing a safe, effective, and clinically translatable protocol for inducing stable tolerance in living donor kidney transplantation. WE HYPOTHESIZE that the ADL+TIS regimen, with modifications deemed necessary to facilitate successful translation to a solid organ transplant setting and with refinements informed by results of the overall U19 program, promotes long-term graft survival in living donor kidney transplant models in NHPs through operational tolerance. To test this hypothesis and facilitate the clinical translation of ADL+TIS, we propose two SPECIFIC AIMS: AIM #1: To determine the efficacy and mechanisms of ADL infusions combined with transient immunosuppression in inducing and maintaining tolerance in a NHP renal transplant model Studies determining the efficacy of the ADL+TIS protocol in achieving operational tolerance of renal allografts in NHPs will be accompanied by deep immune profiling of blood, graft, spleen, and urine to investigate the effects of the protocol on the abundance and activation profiles of distinct effector, exhausted, and regulatory immune cell subsets in various compartments and their spatial organization within graft and spleen. AIM #2: To study the efficacy and mechanisms of inflammasome inhibition and IL-1b antagonism to promote renal transplant tolerance induced by ADL infusions and transient immunosuppression in NHPs Studies in this Aim will determine the ability of strategies inhibiting the inflammasome and its products to synergize with ADL+TIS in promoting operational tolerance in a renal transplant model in NHPs. Mechanistic studies will investigate how inflammasome inhibition and IL-1b antagonism modify the effects of ADLs + TIS on frequencies and activation profiles of myeloid and lymphoid cell subsets in blood, urine, and spleen and their recruitment to and spatial interaction within coordinated immune regulation domains in the renal allograft. The SIGNIFICANCE & INNOVATION of the proposal lie in the efficacy of the ADL+TIS protocol to deplete and exhaust allospecific T cells and to create potent immune regulation. The prospects for tolerance induction to renal transplants in NHPs are high, based on documented tolerance to renal transplants in rodents and islet transplants in NHPs and opportunities for rational refinements of the strategy created by system-level immune profiling.