MCMV infection, latency and reactivation in transplantation tolerance

项目3 项目摘要 在器官捐献者和器官接受者中CMV血清阳性的高流行率使得CMV再激活 移植后的发病率和死亡率的一个重要原因和一个贡献者差移植结果。 CMV再激活的一个新概念指出，移植诱导的炎症是CMV再激活的早期触发因素。 CMV转录再激活。免疫抑制剂的施用进一步抑制病毒特异性 免疫，导致最终完成裂解病毒复制。我们假设，实现强大的 供体特异性移植耐受将抑制移植相关炎症，因此消除了关键的 CMV病毒基因组的表观遗传重编程和转录再激活的早期触发，和 从而防止CMV再激活。我们建议在一个前瞻性的，高度 使用移植前供体“阴性”策略的临床相关供体特异性移植耐受模型 通过递送用化学交联剂1-乙基-3-（3-二甲基氨基丙基）- 碳二亚胺（ECDI）。我们在移植耐受小鼠模型中的研究已经导致了正在进行的 异基因和异种移植的非人灵长类动物模型的临床前研究。而且 基于相同原理的首次人体临床试验，使用肽偶联自体细胞治疗 多发性硬化症最近确立了这种新的耐受性的临床可行性、耐受性和安全性 战略我们使用MCMV的初步数据表明，这种方法的耐受性防止了 潜伏MCMV立即早期（IE）基因转录再激活。相反，急性MCMV感染 受损的尝试耐受诱导和不稳定的建立的耐受。因此，在目前 应用（项目3），我们建议使用小鼠移植模型检查以下三个领域， MCMV感染与ECDI-供者细胞耐受策略：（1）供者特异性耐受对MCMV的影响 急性感染、潜伏期的建立和潜伏期的重新激活;（2）MCMV感染对 供体特异性耐受的诱导和稳定性;（3）相互作用的细胞机制 MCMV感染和供体特异性耐受之间的相互作用。我们的长期目标是确定 在建立和维持稳定的供体特异性的同时预防CMV再活化的治疗靶点 移植耐受性

Project 3 PROJECT SUMMARY A high prevalence of CMV seropositivity in both organ donors and organ recipients makes CMV reactivation following transplantation a significant cause for morbidity and mortality and a contributor to poor graft outcome. An emerging concept in CMV reactivation points to transplant-induced inflammation as an early trigger for CMV transcriptional reactivation. Administration of immunosuppressants further suppresses viral-specific immunity, leading to the eventual completion of lytic viral replication. We hypothesize that achieving robust donor-specific transplant tolerance will inhibit transplant-associated inflammation, therefore remove a critical early trigger for epigenetic reprogramming and transcriptional reactivation of the CMV viral genome, and consequently prevent CMV reactivation. We propose to test this hypothesis in a forward-thinking, highly clinically relevant donor-specific transplant tolerance model using a strategy of pre-transplant donor “negative vaccination” by delivery of donor cells treated with the chemical crosslinker 1-ethyl-3-(3-dimethylaminopropyl)- carbodiimide (ECDI). Our studies in murine models of transplantation tolerance have already led to ongoing pre-clinical studies in non-human primate models of allogeneic and xenogeneic transplantation. Moreover, a first-in-human clinical trial based on the same principle using peptide-coupled autologous cells in patients with multiple sclerosis recently established the clinical feasibility, tolerability, and safety of this novel tolerance strategy. Our preliminary data using MCMV demonstrated that tolerance by this approach prevented immediate early (IE) gene transcriptional reactivation from latent MCMV. Conversely, acute MCMV infection impaired attempted tolerance induction and destabilized established tolerance. Therefore, in the current application (Project 3), we propose to examine the following three areas using murine transplant models with MCMV infection and ECDI-donor cell tolerance strategy: (1) the effects of donor-specific tolerance on MCMV acute infection, establishment of latency, and reactivation from latency; (2) the effects of MCMV infection on the induction and stability of donor-specific tolerance; (3) the cellular mechanisms underlying the reciprocal interactions between MCMV infection and donor-specific tolerance. Our long-term goal is to determine therapeutic targets for prevention of CMV reactivation while establishing and maintaining stable donor-specific transplant tolerance.