Regulatory dendritic cell therapy, promotion of tolerance and underlying mechanisms in NHP renal transplantation

独一无二的是，使用包含共刺激阻断的短期最小免疫抑制(IS)药物方案 (COSB；CTLA4Ig/belatacept)+锥形雷帕霉素，我们已经证明了抗成熟、供体来源的 移植前1周输注DCreg，可以安全地延长恒河猴肾移植存活时间， 伴随着供体反应性记忆T细胞(TMEM)反应的选择性减弱，一种机制 可能有助于克服在NHP和人类中诱导移植耐受的关键障碍。 我们现在将确定一种新的、经修改的、不含CNI的方案是否是(I)更允许延长的方案 移植物存活和(Ii)我们假设将增强DCreg的免疫调节功能，可以实现 供者特有的耐受性。最近有证据表明，淋巴细胞耗尽后会出现COSB(Belatacept) 而雷帕霉素的维持(我们用来在猴子身上证明DCreg疗效的2种药物方案)可以 控制移植患者对COSB的抵抗性排斥反应，减少对COSB(Belatacept)耐药的TMEM。 然而，操作容忍度并未达到。值得注意的是，联合供体DCreg输注(一周 移植前)，围手术期淋巴滤除，促进永久的，供者特异性的同种异体移植存活 啮齿动物。这表明，DCreg输注后的淋巴枯竭不会干扰他们的治疗 效果。此外，移植后淋巴枯竭后输注DCreg，也可促进不确定移植物 生死存亡。因此，我们假设，给ATG注射抗成熟恒河猴DCreg- 淋巴耗竭、CoSb和雷帕霉素治疗的肾移植受者，将导致免疫学变化和 选择性减弱供体反应性TMEM有利于供体特异性耐受。我们进一步假设 宿主同种异体反应性TMEM反应的新生物标记物分析(特别是其Eome的表达) 将与IS的安全撤军相关，并对其进行预测。我们的具体目标是： 目的1：确定供体来源的DCreg移植前输注对移植肾的影响 在给予联合淋巴滤除(ATG)、贝拉泰普和雷帕霉素(ABR)的情况下，NHP的存活率。 目的2：确定供体来源的DCreg移植前输注对移植肾的影响 联合给予ATG、非耗竭型αCD40mAb和雷帕霉素(AAR)的NHP患者存活。 目的3：比较供体和受体来源的DCreg移植后对心脏功能的影响。 联合给予ATG、COSB和雷帕霉素(ABR或AAR)的NHP患者移植肾存活。 每个目标都将伴随着全面的、合理设计的机制研究，这些研究将阐明 过继转移的DCreg和基础蛋白的贩运、命运和免疫调节功能 机械装置。

Uniquely, using a short-term minimal immunosuppressive (IS) drug regimen comprising costimulation blockade (CoSB; CTLA4Ig/belatacept) + tapered rapamycin, we have shown that maturation-resistant, donor-derived DCreg, infused 1w before transplant, can safely prolong renal allograft survival in rhesus macaques, accompanied by selective attenuation of donor-reactive memory T cell (Tmem) responses, a mechanism that may help overcome a critical barrier to transplant tolerance induction in NHP and humans. We will now ascertain whether a novel, modified, CNI-free IS regimen that is (i) more permissive to extended graft survival and (ii) that we hypothesize will enhance the immunomodulatory function of DCreg, can achieve donor-specific tolerance. There is recent evidence that lymphocyte depletion followed by CoSB (belatacept) and rapamycin maintenance (the 2-drug regimen we used to demonstrate DCreg efficacy in monkeys) can control CoSB-resistant rejection in transplant patients, with reduction in CoSB (belatacept)-resistant Tmem. However, operational tolerance was not achieved. Notably, combination of donor DCreg infusion (a week before transplant) with perioperative lymphodepletion, promotes permanent, donor-specific allograft survival in rodents. This indicates that lymphodepletion after DCreg infusion does not interfere with their therapeutic effect. Moreover, DCreg infusion post-transplant following lymphodepletion, can also promote indefinite graft survival. We therefore hypothesize that maturation-resistant rhesus DCreg, administered to ATG- lymphodepleted, CoSB and rapamycin-treated renal graft recipients, will induce immunological changes and selective attenuation of donor-reactive Tmem conducive to donor-specific tolerance. We further hypothesize that novel biomarker analyses of host alloreactive Tmem responses (in particular, their expression of Eomes) will correlate with and be predictive of safe withdrawal of IS. Our Specific Aims are: Aim 1: To determine the influence of donor-derived DCreg infusion before transplant on renal allograft survival in NHP given combined lymphodepletion (ATG), belatacept and rapamycin (ABR). Aim 2: To determine the influence of donor-derived DCreg infusion before transplant on renal allograft survival in NHP given combined ATG, non-depleting αCD40 mAb and rapamycin (AAR). Aim 3: To compare the influence of donor-versus recipient-derived DCreg infusion post-transplant on renal allograft survival in NHP given combined ATG, CoSB and rapamycin (ABR or AAR). Each Aim will be accompanied by comprehensive, rationally-designed mechanistic studies that will elucidate the trafficking, fate and immune regulatory function of the adoptively-transferred DCreg and underlying mechanisms.