Productive lytic replication of a recombinant Kaposi's sarcoma-associated herpesvirus in efficient primary infection of primary human endothelial cells

被引:104
作者
Gao, SJ
Deng, JH
Zhou, FC
机构
[1] Univ Texas, Hlth Sci Ctr, Dept Pediat, San Antonio, TX 78229 USA
[2] Univ Texas, Hlth Sci Ctr, Childrens Canc Res Inst, Tumor Virol Program, San Antonio, TX 78229 USA
[3] Univ Texas, Hlth Sci Ctr, Dept Immunol & Microbiol, San Antonio, TX 78229 USA
[4] Univ Texas, Hlth Sci Ctr, Dept Med, San Antonio, TX 78229 USA
[5] San Antonio Canc Inst, San Antonio, TX 78229 USA
关键词
D O I
10.1128/JVI.77.18.9738-9749.2003
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Kaposi's sarcoma-associated herpesvirus (KSHV) is linked to the development of Kaposi's sarcoma (KS), a vascular spindle cell tumor primarily consisting of proliferating endothelial cells. Although KSHV has been shown to infect primary human endothelial cells and convert them into spindle shapes, KSHV infection is largely latent, and efforts to establish a highly efficient and sustainable infection system have been unsuccessful. A recombinant KSHV, BAC36, that has high primary-infection efficiency in 293 cells has been obtained (F. C. Zhou, Y. J. Zhang, J. H. Deng, X. P. Wang, H. Y. Pan, E. Hettler, and S. J. Gao, J. Virol. 76:6185-6196, 2002). BAC36 contains a green fluorescent protein cassette which can be used to conveniently monitor viral infection. Here, we describe the establishment of a KSHV lytic-replication-permissive infection cell model using BAC36 virions to infect primary human umbilical vein endothelial cell (HUVEC) cultures. BAC36 infection of HUVEC cultures has as high as 90% primary-infection efficiency and consists of two phases: a permissive phase, in which the cultures undergo active viral lytic replication, producing a large number of virions and concomitantly resulting in large-scale cell death, and a latent phase, in which the surviving cells from the permissive phase switch into latent infection, with a small number of cells undergoing spontaneous viral lytic replication, and proliferate into bundles of spindle cells with KS slit-like spaces. An assay for determining the KSHV titer in a virus preparation has also been developed. The cell model should be useful for examining KSHV infection and replication, as well as for understanding the development of KS.
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页码:9738 / 9749
页数:12
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