Regulatory T cells to promote mixed chimerism for tolerance to islets and kidneys from deceased and living donors

这项U19提案的目的是通过两种细胞的混合嵌合体来实现胰岛和肾脏移植耐受。 补充食蟹猴项目，由三个核心支持。这两个项目都有相关性 终末期肾病（ESRD）和1型糖尿病（T1 D）的治愈性治疗。他们雇佣相关人员 分别实现对死亡和活体供体移植物的耐受性的方法。项目1旨在利用 扩增的多克隆受体T细胞，以开发压缩的预处理方案， 同时进行骨髓、肾脏和胰岛的同种异体移植我们以前用过6- 仅适用于活体供体的日间预处理方案，以实现短暂的嵌合体和肾脏 猴子和人类的同种异体移植耐受性。我们现在的目标是制定一个条件协议， 在确认死亡供体后，允许在24小时内进行移植（Tx）。我们的方法建立在 我们证明，扩增的多克隆受体THBE可以实现更长的嵌合体， 比之前在食蟹猴模型中可能的耐受性更强。宽容避免了 长期免疫抑制治疗对胰岛移植物有破坏作用。此外，我们在NOD的研究 小鼠已经显示，持久混合嵌合体的非清髓性诱导逆转了晚期抗胰岛细胞抗体， 自身免疫，同时避免目前限制免疫的同种异体免疫、自身免疫和药物毒性。 胰岛Tx在T1 D中的功效。项目2旨在开发一种耐受诱导策略，用于治愈性治疗 使用活体亲属供体（LRD）复合胰岛-肾（IK）Tx.项目 基于我们的观察，在大型动物中移植预血管化的胰岛作为复合IKs的一部分， 模型需要比游离的非血管化胰岛的Tx少得多的胰岛来实现胰岛素独立性。我们 最近在恒河猴中使用一种新的低强度造血细胞移植实现了IKs的耐受性 “父母对子女”组合的协议。项目2旨在调整预处理方案的组成部分 和/或可能对胰岛功能具有早期负面影响的供体细胞来源。利用了 为了在LRD Tx之前产生有效的供体特异性TCLs，我们将测试这些细胞促进持久免疫的能力。 混合异基因嵌合体，具有逆转T1 D的潜力。这两个项目将包括广泛的机械 基于高通量TCR测序方法的分析，用于追踪同种异体反应性T细胞 我们已经在人类中开发了这些库，并将在核心B中应用于食蟹猴。因此，我们旨在 在两个项目中实现持久的混合嵌合体，以治愈自身免疫，同时预防 通过诱导耐受性的同种免疫攻击。核心A将为两个项目提供猴岛，核心B将 开发高通量T细胞受体（TCR）测序平台，用于识别和跟踪 供体特异性同种异体反应性T细胞克隆的命运，提供了一个独特的机制工具，适用于这两个项目。 核心C将为所有项目和核心提供行政、生物储存和数据管理支持。

This U19 proposal aims to achieve islet and kidney allograft tolerance through mixed chimerism in two complementary cynomolgus monkey projects that are supported by three cores. Both projects have relevance for end-stage renal disease (ESRD) and curative therapy for Type 1 diabetes (T1D). They employ related approaches to achieve tolerance for deceased and living donor grafts, respectively. Project 1 aims to use expanded polyclonal recipient Tregs to develop a compressed conditioning regimen that induces tolerance of simultaneous bone marrow, kidney and islet allografts from deceased donors. We have previously used a 6- day conditioning protocol that is relevant only for living donors, to achieve transient chimerism and kidney allograft tolerance in monkeys and humans. We now aim to develop a conditioning protocol that could be initiated after identification of a deceased donor, allowing transplantation (Tx) within 24 hours. Our approach builds on our demonstration that expanded polyclonal recipient Tregs can achieve far more prolonged chimerism and more robust tolerance than has previously been possible in the cynomolgus model. Tolerance obviates the need for long-term immunosuppressive therapy with its destructive effects on islet grafts. Moreover, our studies in NOD mice have shown that non-myeloablative induction of durable mixed chimerism reverses advanced anti-islet autoimmunity, simultaneously avoiding the alloimmunity, autoimmunity and drug toxicity that currently limit the efficacy of islet Tx in T1D. Project 2 aims to develop a tolerance induction strategy for curative treatment of end-stage diabetic nephropathy using living related donor (LRD) composite Islet-Kidney (IK) Tx. The project builds on our observation that transplanting pre-vascularized islets as part of composite IKs in large animal models requires far fewer islets to achieve insulin independence than Tx of free, non-vascularized islets. We recently achieved tolerance of IKs in rhesus monkeys using a novel, low intensity, hematopoietic cell transplant protocol in a “parent-to-offspring” combination. Project 2 aims to adjust components of the conditioning regimen and/or donor cell source that may have an early negative impact on islet function. Taking advantage of the ability to generate potent donor-specific Tregs prior to LRD Tx, we will test the ability of these cells to promote durable mixed allogeneic chimerism, with its potential to reverse T1D. Both projects will include extensive mechanistic analyses that build on a high-throughput TCR sequencing-based approach for tracking the alloreactive T cell repertoire that we have developed in humans and will apply to cynomolgus monkeys in Core B. Thus, we aim to achieve durable mixed chimerism in both projects to cure autoimmunity while simultaneously preventing alloimmune attack by inducing tolerance. Core A will provide monkey islets for both projects and Core B will develop a high throughput T cell receptor (TCR) sequencing platform that will be used to identify and track the fate of donor-specific alloreactive T cell clones, providing a unique mechanistic tool to be applied in both projects. Core C will provide administrative, biorepository and data management support for all of the projects and cores.