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Journal Article Centrifugal enhancement of Kaposi's sarcoma-associated virus infection of human endothelial cells in vitro
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Authors
Seung-Min Yoo, Ae-Kyung Ahn, Tae Gun Seo, Hyo Bong Hong, Myung-Ae Chung, Sang-Don Jung, Hae Wol Cho, Myung-Shin Lee
Issue Date
2008-12
Citation
Journal of Virological Methods, v.154, no.1-2, pp.160-166
ISSN
0166-0934
Publisher
Elsevier
Language
English
Type
Journal Article
DOI
https://dx.doi.org/10.1016/j.jviromet.2008.07.026
Abstract
In order to improve the efficiency of infection of primary human endothelial cells in vitro of Kaposi's sarcoma-associated herpesvirus (KSHV), the effect of low speed centrifugation was investigated. The recombinant KSHV, BAC36, was used to examine the centrifugal enhancement of KSHV. Infectivity was estimated by green fluorescent protein (GFP) expression and real-time RT-PCR. The enhancement of infectivity was dependent upon the time and force of centrifugation in human umbilical vein endothelial cells (HUVECs). Centrifugation enhanced the infectivity of KSHV by up to 70 fold compared to non-centrifugal control infection for the same period of time; viral mRNA expression was also enhanced by centrifugation. HUVECs that were centrifuged before infection with KSHV displayed no enhancement in infectivity; therefore, enhancement is believed to occur during centrifugation. In addition, the mechanisms of infection including the initial viral attachment to cells, lipid rafts, and clathrin-mediated and caveolae endocytosis appear to be similar in KSHV infection with and without centrifugal enhancement. These results show that low speed centrifugation could be a useful tool for improving the efficiency of KSHV infection in vitro. © 2008 Elsevier B.V. All rights reserved.
KSP Keywords
Human endothelial cells, Kaposi's sarcoma, MRNA expression, Real-time RT-PCR, green fluorescent protein, human umbilical vein endothelial cells, lipid rafts, low speed, virus infection