Human cytomegalovirus entry into epithelial and endothelial cells depends on genes UL128 to UL150 and occurs by endocytosis and low-pH fusion
Human cytomegalovirus entry into epithelial and endothelial cells depends on genes UL128 to UL150 and occurs by endocytosis and low-pH fusion
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DOI：
10.1128/jvi.80.2.710-722.2006
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发表时间：
2006-01-01
影响因子：
5.4
通讯作者：
Johnson, DC
中科院分区：
文献类型：
--
作者：
Ryckman, BJ;Jarvis, MA;Johnson, DC
人巨细胞病毒（HCMV）在上皮细胞和内皮细胞中的复制似乎在病毒传播、疾病和持久性中很重要。研究这些细胞类型的感染是困难的，因为HCNW实验室菌株（例如，AD 169和Towne）在成纤维细胞中广泛增殖期间失去了感染培养的上皮细胞和内皮细胞的能力。HCMV的临床菌株（例如，TR和FIX）具有在实验室菌株中大量突变的基因簇（UL 128至UL 150），并且最近的研究表明这些基因促进上皮和内皮细胞中的复制。这些基因促进这两种细胞类型感染的机制尚不清楚。我们从TR菌株TR Delta 4中获得了HCMV UL 128-UL 150缺失突变体，并研究了HCMV感染上皮细胞和内皮细胞的早期事件以及基因UL 128至UL 150的作用。野生型TR的分析表明，HCMV进入上皮细胞和内皮细胞的内吞作用，然后低pH值依赖性融合，这是不同的pH值无关的融合与质膜观察与人成纤维细胞。TR Delta 4在早期感染过程中显示出许多缺陷。与TR相比，TR Delta 4在上皮细胞上的吸附和进入较差，但这些缺陷可以通过更高剂量的病毒和使用聚乙二醇（PEG）促进病毒体与细胞膜之间的融合来克服。高多样性和PEG处理没有促进TR Delta 4感染内皮细胞，但病毒颗粒被内化。总之，这些数据表明基因UL 128至UL 150是HCMV吸附和穿透上皮细胞所需的，并在病毒进入内皮细胞后促进病毒复制的某些早期阶段。
Human cytomegalovirus (HCMV) replication in epithelial and endothelial cells appears to be important in virus spread, disease, and persistence. It has been difficult to study infection of these cell types because HCNW laboratory strains (e.g., AD169 and Towne) have lost their ability to infect cultured epithelial and endothelial cells during extensive propagation in fibroblasts. Clinical strains of HCMV (e.g., TR and FIX) possess a cluster of genes (UL128 to UL150) that are largely mutated in laboratory strains, and recent studies have indicated that these genes facilitate replication in epithelial and endothelial cells. The mechanisms by which these genes promote infection of these two cell types are unclear. We derived an HCMV UL128-to-UL150 deletion mutant from strain TR, TR Delta 4, and studied early events in HCMV infection of epithelial and endothelial cells, and the role of genes UL128 to UL150. Analysis of wild-type TR indicated that HCMV enters epithelial and endothelial cells by endocytosis followed by low-pH-dependent fusion, which is different from the pH-independent fusion with the plasma membrane observed with human fibroblasts. TR Delta 4 displayed a number of defects in early infection processes. Adsorption and entry of TR Delta 4 on epithelial cells were poor compared with those of TR, but these defects could be overcome with higher doses of virus and the use of polyethylene glycol (PEG) to promote fusion between virion and cellular membranes. High multiplicity and PEG treatment did not promote infection of endothelial cells by TR Delta 4, yet virus particles were internalized. Together, these data indicate that genes UL128 to UL150 are required for HCMV adsorption and penetration of epithelial cells and to promote some early stage of virus replication, subsequent to virus entry, in endothelial cells.