Enhanced green fluorescent protein as a marker for localizing murine cytomegalovirus in acute and latent infection
Enhanced green fluorescent protein as a marker for localizing murine cytomegalovirus in acute and latent infection
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DOI：
10.1016/s0166-0934(00)00202-0
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发表时间：
2000-09-01
影响因子：
3.1
通讯作者：
Hamilton, JD
中科院分区：
文献类型：
--
作者：
Henry, SC;Schmader, K;Hamilton, JD
构建了表达增强型绿色荧光蛋白(EGFP)的重组小鼠巨细胞病毒(MCMV)，可直接检测潜伏感染和再激活感染中的病毒活性部位。重组病毒具有与野生型MCMV相似的急性和潜伏感染特征。在潜伏感染的肺、脾石蜡切片上可见少见的绿色荧光灶。同一肺组织切片中EGFP免疫过氧化物酶阳性染色提示，这些细胞可能是病毒基因表达受限的部位。在体外潜伏感染复活的组织外植体中容易检测到绿色荧光蛋白。荧光细胞的形态和黏附特性表明，病毒在外植体培养中的组织巨噬细胞中发生了重新激活。这项研究中提出的观察结果表明，表达EGFP的重组体作为直接跟踪MCMV体内和体外活性的工具是有用的。(C)2000 Elsevier Science B.V.保留所有权利。
A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteristics similar to those of wild-type mCMV. Rare green-fluorescing foci were observed in paraffin sections from lungs and spleens infected latently. Positive immunoperoxidase staining for EGFP in sections of the same lung tissues suggests that these cells may be sites of restricted viral gene expression. EGFP was detected easily in tissue explants reactivating from latent infection in vitro. Morphology and adhesion characteristics of fluorescing cells suggest that viral reactivation occurs in tissue macrophages in explant cultures. The observations presented in this study demonstrate the usefulness of EGFP-expressing recombinants as tools for direct tracking of mCMV activity in vivo and in vitro. (C) 2000 Elsevier Science B.V. All rights reserved.