LATENCY AND REACTIVATION OF CYTOMEGALOVIRUS AFTER BONE MARROW TRANSPLANTATION

巨细胞病毒(CMV)的终生潜伏期提供了一个蓄水池 会增加疾病风险，无论是与来自 供者或在骨髓(BM)接受者体内重新激活。一点儿 已知决定传播风险的参数或 重新激活。在太平洋标准时间的四年里，对自然界的调查 人类巨细胞病毒：造血祖细胞的相互作用导致 潜伏基因产物的发现和表征。这 新项目将专注于研究潜伏基因的表达 在自然宿主中的产物，并将评估表达作为预测因素 巨细胞病毒的传播和重新激活。首先，这项工作将确定 CMV在自然界潜伏期驻留的细胞类型(S) 宿主，紧随CD34+和CD33+造血细胞。bm 健康血清阳性献血者(或外周血)骨髓单个核细胞 从骨髓动员的GM-CSF的血液单个核细胞)将 进行流式细胞仪分析，确定哪种细胞类型(S)最多 经常藏有病毒基因组，含有潜伏的转录本和 表达潜伏期相关蛋白。第二，补充研究 关于潜伏细胞类型，该项目将评估基因模式 表达作为CMV传播的预测因子和指标 同种异体移植过程中的再激活。骨髓和外周血 将对单个核细胞进行转录模式评估， 区分急性感染和潜伏感染。Sera将会是 评估抗体和淋巴细胞将评估 能够被潜伏期和潜伏期表达的蛋白质刺激 生殖性感染。血清阳性献血者的表达模式将 与血清阴性受者的结局和模式相关 在血清阳性的接受者中将与可能性相关 它们会经历重新激活的感染。第三，本项目将评估 健康血清阳性骨髓祖细胞培养后的再激活 捐赠者使用我们发现的重新激活的培养条件 CD33+、CD15+、CD10+细胞中CMV潜伏感染的实验研究 CD1a+细胞。能够产生重新激活的病毒的细胞 将通过流式细胞仪分离病毒抗原阳性进行鉴定 细胞，通过分析是否存在与 产生性感染和产生病毒的空斑试验。这个 我们建议研究的事件和我们建议研究的细胞类型 专注似乎有助于终生坚持和潜伏 在自然感染的宿主中感染CMV，而这个储存库可能 使骨髓移植受者易于重新激活和传播 活动性巨细胞病毒感染

Lifelong latency of cytomegalovirus (CMV) provides a reservoir that contributes to disease risk, whether transferred with cells from the donor or reactivated within the bone marrow (BM) recipient. Little is known of the parameters that determine risk of transmission or reactivation. Over the pst four years investigations into the nature of the human CMV:hematopoietic progenitor cell interactions have led to the discovery and characterization of latent gene products. This new project will focus on investigating the expression of latent gene products in the natural host and will assess expression as a predictor of CMV transmission and reactivation. First, this work will identify the cell type(s) in which CMV resides during latency in the natural host, following CD34+ and CD33+ hematopoietic cells most closely. BM mononuclear cells from BM of health seropositive donors (or peripheral blood mononuclear cells from GM-CSF mobilized from BM) will be subjected to FACS analysis to determine which cell type(s) most frequently harbor viral genomes, contain latent transcripts and express the latency-associated protein. Second, to complement studies on the latent cell type, the project will assess the pattern of gene expression as a predictor of CMV transmission and as an indicator of reactivation during allograft transplantation. BM and peripheral blood mononuclear cells will be evaluated for transcription patterns which distinguish between acute and latent infection. Sera will be evaluated for antibody to and lymphocytes will be evaluated for ability to be stimulated by proteins expressed during latent and productive infection. Expression patterns in seropositive donors will be correlated with outcome in the seronegative recipient, and patterns in the seropositive recipient will be correlated with the likelihood they undergo reactivated infection. Third, this project will assess reactivation in BM progenitors cultured from healthy seropositive donors using culture conditions that we have found to reactivate experimental latent CMV infection in cultured CD33+, CD15+, CD10+, CD1A+ cells. Cells that are capable of yielding reactivated virus will be identified through FACS separation for viral antigen-positive cells, by analysis for presence of transcripts consistent with productive infection and by plaque assay for production of virus. The events that we propose to study and the cell types on which we propose to focus appear to contribute to the lifelong persistence and latency of CMV in the naturally infected host, and this reservoir may predispose BM transplant recipients to reactivation and dissemination of active CMV infection