Compton T, Nowlin D M, Cooper N R
Department of Immunology, Scripps Research Institute, La Jolla, California 92037.
Virology. 1993 Apr;193(2):834-41. doi: 10.1006/viro.1993.1192.
In this report, we demonstrate that the initial event in human cytomegalovirus (HCMV) infection is attachment to extracellular heparan sulfate. Further, this interaction is important for initiation of infection in fibroblast cells. Using microbinding assays to specifically monitor virus attachment as well as plaque titration assays to measure infectivity, we found that heparin competition as well as enzymatic digestion of cells with heparinase blocked virus attachment, initiation of immediate-early gene expression and infectivity. Other major glycosaminoglycans were found not to be involved in HCMV attachment and infectivity. In addition, HCMV was unable to attach to mutant derivatives of Chinese hamster ovary cells deficient in synthesis of heparan sulfate proteoglycans. Basic fibroblast growth factor, which requires initial interaction with extracellular heparin prior to binding to its high affinity receptor, also inhibited HCMV attachment to cells. Time-course experiments revealed that the initial HCMV binding was sensitive to heparin competition (10 micrograms/ml) or 0.75 M salt washes. The initial heparin-dissociable binding converted rapidly to high affinity (heparin resistant) HCMV attachment. These data suggest that sequential receptor interactions may mediate HCMV adsorption to cells. Heparin affinity chromatography revealed that multiple HCMV envelope glycoproteins, including gB, are capable of binding to heparin.
在本报告中，我们证明人巨细胞病毒（HCMV）感染的初始事件是与细胞外硫酸乙酰肝素结合。此外，这种相互作用对于成纤维细胞中感染的起始很重要。使用微结合试验特异性监测病毒附着以及蚀斑滴定试验测量感染性，我们发现肝素竞争以及用肝素酶对细胞进行酶消化会阻断病毒附着、立即早期基因表达的起始和感染性。发现其他主要糖胺聚糖不参与HCMV附着和感染性。此外，HCMV无法附着到缺乏硫酸乙酰肝素蛋白聚糖合成的中国仓鼠卵巢细胞的突变衍生物上。碱性成纤维细胞生长因子在与高亲和力受体结合之前需要先与细胞外肝素进行初始相互作用，它也抑制HCMV附着到细胞上。时间进程实验表明，初始的HCMV结合对肝素竞争（10微克/毫升）或0.75M盐洗涤敏感。最初可被肝素解离的结合迅速转变为高亲和力（抗肝素）的HCMV附着。这些数据表明，连续的受体相互作用可能介导HCMV吸附到细胞上。肝素亲和层析显示，包括gB在内的多种HCMV包膜糖蛋白能够与肝素结合。
