Mechanisms of CMV latency in accelerated vascular disease

描述（由申请人提供）：本项目的目的是确定巨细胞病毒（CMV）加速移植血管硬化（TVS）的病毒机制，TVS是与实体器官移植物慢性排斥反应相关的标志性血管疾病。临床研究表明，人巨细胞病毒（HCMV）与移植血管硬化（TVS）和血管成形术后血管再狭窄以及动脉粥样硬化的加速直接相关。我们已经开发了一种大鼠心脏移植慢性排斥模型，该模型表现出人类TVS的所有特征。本课题组和其他研究表明，大鼠巨细胞病毒（RCMV）感染显著加速了大鼠心脏移植模型中TVS和慢性排斥反应的发生。我们已经观察到，与在临床环境中观察到的相似，与未感染的对照组相比，来自潜伏感染供体大鼠的心脏同种异体移植物经历慢性排斥反应的加速。有趣的是，我们还观察到，更昔洛韦治疗，这是一种病毒的DMA聚合酶抑制剂，阻断病毒晚期基因的表达，并没有阻止RCMV潜伏感染的同种异体移植物移植到血清阴性受体的TVS的加速。这些结果表明，完全的病毒复制是不需要的疾病的加速和病毒的表达限于CMV立即早期（IE）或早期基因产物。在这项研究中，我们将确定移植物中感染的病毒载量和细胞类型。此外，我们将确定在移植前和移植后潜伏感染供体的同种异体移植物中表达的病毒基因，然后确定这些基因产物是否有助于加速TVS？为了实现这些目标，我们计划以下具体目标：1.移植前同种异体移植物中RCMV潜伏感染的细胞类型以及病毒表达谱是什么？A.移植前供者心脏中RCMV潜伏感染的细胞类型是什么？B。哪些RCMV基因在潜在感染的供体心脏中表达？2.潜伏感染供体的同种异体移植物中RCMV加速TVS的特征是什么？A.潜伏感染供体的同种异体移植物中TVS形成的动力学是什么？B。在更昔洛韦存在下，潜伏感染的同种异体移植物中RCMV基因表达谱在TVS发展过程中是否发生变化？3.在潜伏感染的同种异体移植物中表达的哪种RCMV基因是加速TVS所必需的？A.潜伏感染的同种异体移植物中表达的RCMV基因突变是否改变病毒诱导的疾病加速？B。在潜伏感染的同种异体移植物中表达的RCMV基因的功能是什么？

DESCRIPTION (provided by applicant): The goal of this project is to determine the viral mechanisms involved in cytomegalovirus (CMV)-accelerated transplant vascular sclerosis (TVS), which is the hallmark vascular disease associated with chronic rejection of solid organ grafts. Clinical studies have directly associated human cytomegalovirus (HCMV) with the acceleration of transplant vascular sclerosis (TVS) and vascular restenosis following angioplasty, as well as atherosclerosis. We have developed a rat heart transplant chronic rejection model that exhibits all of the hallmarks of TVS in humans. Studies by our group and others have shown that rat CMV (RCMV) infection significantly accelerates both the development of TVS as well as chronic rejection in the rat heart allograft model. We have observed that similar to what is observed in the clinical setting that heart allografts from latently infected donor rats undergo acceleration of chronic rejection compared to uninfected controls. Interestingly, we have also observed that treatment with ganciclovir, which is a viral DMA polymerase inhibitor that blocks viral late gene expression, did not prevent the acceleration of TVS in RCMV latently infected allografts transplanted into sero-negative recipients. These results indicate that complete viral replication is not required for the acceleration of disease and that virus expression is restricted to CMV immediate early (IE) or early gene products. In this study, we will determine the viral load and cell types infected in the allograft. In addition, we will determine the viral genes that are expressed in the allografts from latently infected donors prior to and following transplantation and then determine whether these gene products contribute to the acceleration of TVS? To achieve these objectives we plan the following specific aims: 1. What are the RCMV latently infected cell types as well as the viral expression profile in the allograft prior to transplantation? A. What are the RCMV latently infected cell types in the donor hearts prior to transplantation? B. Which RCMV genes are expressed in the latently infected donor hearts? 2. What are the characteristics of RCMV acceleration of TVS in allografts from latently infected donors? A. What are the kinetics of TVS formation in allografts from latently infected donors? B. Does the spectrum of RCMV gene expression change in the latently infected allografts during the development of TVS in the presence of ganciclovir? 3. Which RCMV gene(s) expressed in latently infected allografts are required for the acceleration of TVS? A. Does mutation of the RCMV genes expressed in the latently infected allografts alter virus-induced acceleration of disease? B. What is the function of RCMV genes expressed in latently infected allografts?