LATENCY AND REACTIVATION OF CYTOMEGALOVIRUS AFTER BONE MARROW TRANSPLANTATION

巨细胞病毒（CMV）的终身潜伏期提供了一个储存库， 有助于疾病的风险，无论是从细胞转移， 供体或在骨髓（BM）受体内重新激活。 小 已知确定传播风险的参数，或 重新激活 在过去的四年里， 人类巨细胞病毒：造血祖细胞的相互作用， 潜在基因产物的发现和表征。 这 一个新的项目将集中研究潜在基因的表达 天然宿主中的产物，并将评估表达作为预测因子 CMV的传播和再激活。 首先，这项工作将确定 CMV在自然界中的潜伏期期间驻留的细胞类型 宿主，其次是CD 34+和CD 33+造血细胞。BM 来自健康血清阳性供体（或外周血供体）的BM的单核细胞 来自骨髓动员的GM-CSF的血液单核细胞）， 进行FACS分析，以确定哪种细胞类型最 经常携带病毒基因组，含有潜伏的转录本， 表达潜伏相关蛋白。 第二，补充研究 在潜伏细胞类型上，该项目将评估基因的模式， 表达作为CMV传播的预测因子和作为 在同种异体移植过程中重新激活。BM和外周血 将评估单核细胞的转录模式， 区分急性感染和潜伏感染。 塞拉会 将评价抗体和淋巴细胞 能够被潜伏期表达的蛋白质刺激， 生产性感染 血清阳性供体的表达模式将 与血清阴性受体的结果和模式相关 在血清反应阳性的接受者中， 它们会经历重新激活的感染。 第三，该项目将评估 在从健康血清反应阳性细胞培养的BM祖细胞中的再活化 使用我们发现的培养条件， 培养的CD 33+，CD 15+，CD 10+， CD 1A+细胞。 能够产生再活化病毒的细胞 将通过FACS分离确定病毒抗原阳性 细胞，通过分析是否存在与 生产性感染和噬斑测定病毒的产生。 的 我们建议研究的事件和我们建议研究的细胞类型 集中注意力似乎有助于终身的持久性和潜伏期 CMV在自然感染的宿主中，这个水库可能 使骨髓移植受者易于再活化和播散 活动性CMV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