Apoptotic Donor Leukocytes to Promote Kidney Transplant Tolerance

项目2总结 我们证明了两种移植周输注的凋亡性供者白细胞(ADL)和一过性 α-CD40、雷帕霉素、sTNFR和α-IL-6R联合免疫抑制可诱导长期(1年)耐受 对5个非致敏、1个MHC-II DRB等位基因相合的非人灵长类动物中的5个进行胰岛移植。项目2 将研究ADL+TIS方案在肾移植模型中的疗效和机制，并应用高密度的 维度免疫分析以指导方案改进，其主要目标是开发一种安全的、 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，经过必要的修改以促进成功 翻译到坚实的器官移植环境，并通过整体U19计划的结果进行改进， 通过手术耐受促进NHP活体供肾移植模型中移植物的长期存活。 为了验证这一假说并促进ADL+TIS的临床翻译，我们提出了两个具体的目标： 目的#1：确定ADL输注联合瞬变的疗效和机制 免疫抑制在诱导和维持NHP肾移植耐受中的作用 ADL+TIS方案在实现同种异体肾移植手术耐受中的有效性研究 NHPS将伴随着对血液、移植物、脾和尿液的深度免疫分析来研究其影响。 关于不同效应性、疲劳性和调节性免疫的丰度和激活曲线的方案 移植物和脾内各隔室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拮抗作用及其机制。 输注ADL和短暂性免疫抑制促进肾移植耐受 这一目标的研究将确定抑制炎症体及其产品的策略的能力 与ADL+TIS协同作用促进NHP肾移植模型的手术耐受性。机械论 研究将探讨炎性小体抑制和IL-1b拮抗如何改变ADLS+TIS的作用 血液、尿液和脾中髓系细胞和淋巴细胞亚群的频率和激活谱及其相互关系 肾移植中协调免疫调节域的募集和空间相互作用。 该方案的意义和创新之处在于ADL+TIS议定书对耗尽和排气的有效性 同种异体特异性T细胞，并产生有效的免疫调节。肾耐受诱导的前景 根据记录的啮齿动物和胰岛移植对肾移植的耐受性，NHP中的移植比例很高。 在NHP中，以及通过系统级免疫图谱创建的合理改进策略的机会。

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.