Experimental human cytomegalovirus latency in CD14+ monocytes
Experimental human cytomegalovirus latency in CD14+ monocytes
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DOI：
10.1073/pnas.1014509107
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发表时间：
2010-11-16
影响因子：
11.1
通讯作者：
Shenk, Thomas E.
中科院分区：
文献类型：
--
作者：
Hargett, Danna;Shenk, Thomas E.
CD14（+）单核细胞是潜伏的人类巨细胞病毒的储存库，在向巨噬细胞或树突状细胞分化的过程中，病毒复制被重新激活。目前尚不清楚病毒是否可以在直接感染单核细胞时建立潜伏期，或者它是否必须首先在祖细胞中处于静止状态，然后分化产生单核细胞。我们报告，原代人单核细胞感染与人类巨细胞病毒的临床株表现出潜伏期的特点。我们建立了防止单核细胞分化至少25天的培养条件，正如细胞表面标记物表达所证明的那样。用FIX临床菌株感染这些单核细胞导致许多病毒裂解rna的短暂积累和四种先前描述的潜伏期相关转录物的持续表达。病毒DNA的数量在感染后保持不变，细胞表面和总HLA-DR蛋白在感染后持续大幅减少。当使用刺激分化为巨噬细胞或树突状细胞表型的细胞因子混合物处理时，受感染的单核细胞重新激活病毒复制并产生感染性后代。单独用IL-6处理受感染的单核细胞也足以重新激活，暴露于这种细胞因子后产生的颗粒的传染性比其他处理产生的病毒粒子高约5倍。我们认为，体内微环境不仅影响再激活的效率，而且影响潜伏感染的单核细胞产生的病毒粒子的感染性。
CD14(+) monocytes are a reservoir for latent human cytomegalovirus, and virus replication is reactivated during their differentiation to macrophages or dendritic cells. It has not been clear whether the virus can establish latency upon direct infection of monocytes or whether it must first become quiescent in a progenitor cell that subsequently differentiates to generate a monocyte. We report that infection of primary human monocytes with a clinical strain of human cytomegalovirus exhibits the hallmarks of latency. We established conditions for culturing monocytes that prevent differentiation for at least 25 d, as evidenced by cell surface marker expression. Infection of these monocytes with the FIX clinical strain resulted in transient accumulation of many viral lytic RNAs and sustained expression of four previously described latency-associated transcripts. The amount of viral DNA remained constant after infection, and cell surface and total HLA-DR proteins were substantially reduced on a continuing basis after infection. When treated with cytokine mixtures that stimulate differentiation to a macrophage or dendritic cell phenotype, infected monocytes reactivated virus replication and produced infectious progeny. Treatment of infected monocytes with IL-6 alone also was sufficient for reactivation, and the particles produced after exposure to this cytokine were about fivefold more infectious than virions produced by other treatments. We propose that in vivo microenvironments influence not only the efficiency of reactivation but also the infectivity of the virions produced from latently infected monocytes.