Renne R, Blackbourn D, Whitby D, Levy J, Ganem D
Howard Hughes Medical Institute and Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California 94143-0414, USA.
J Virol. 1998 Jun;72(6):5182-8. doi: 10.1128/JVI.72.6.5182-5188.1998.
Kaposi's sarcoma-associated herpesvirus (KSHV) (also called human herpesvirus 8) is a novel gammaherpesvirus strongly implicated in the pathogenesis of Kaposi's sarcoma. Although virions can be produced in high yield from latently infected B-cell lines treated with phorbol esters, little is known about the infectivity of such virus, and efficient serial propagation of KSHV has been problematic. Here we report on the infectivity of KSHV produced from phorbol-induced BCBL-1 cells, employing an assay based on the detection of a spliced late mRNA by a sensitive reverse transcriptase PCR (RT-PCR) method. The results of this study confirm previous observations that 293 cells are susceptible to viral infection; however, infection with BCBL-1-derived virus is inefficient and the pattern of viral gene expression in infected cells may not fully reproduce that of authentic lytic infection. In keeping with this finding, serial propagation of BCBL-1-derived virus could not be demonstrated on 293 cells. Eleven of 38 other cell lines tested also supported KSHV infection, as judged by this RT-PCR assay, including cells of B-cell, endothelial, epithelial, and fibroblastic origin; however, in all cases, infection proceeded at or below the levels observed in 293 cells.
卡波西肉瘤相关疱疹病毒(KSHV)(也称为人类疱疹病毒8型)是一种新型γ疱疹病毒,与卡波西肉瘤的发病机制密切相关。尽管用佛波酯处理潜伏感染的B细胞系可大量产生病毒粒子,但对于此类病毒的感染性知之甚少,而且KSHV的有效连续传代一直存在问题。在此,我们利用一种基于通过灵敏的逆转录酶PCR(RT-PCR)方法检测剪接晚期mRNA的试验,报告了从佛波酯诱导的BCBL-1细胞产生的KSHV的感染性。本研究结果证实了先前的观察结果,即293细胞易受病毒感染;然而,用BCBL-1衍生病毒感染效率低下,且感染细胞中病毒基因表达模式可能无法完全重现真正的裂解感染模式。与此发现一致的是,在293细胞上无法证明BCBL-1衍生病毒的连续传代。通过该RT-PCR试验判断,所测试的其他38种细胞系中有11种也支持KSHV感染,包括B细胞、内皮细胞、上皮细胞和成纤维细胞来源的细胞;然而,在所有情况下,感染程度均等于或低于在293细胞中观察到的水平。