Centrifugal enhancement of Kaposi's sarcoma-associated virus infection of human endothelial cells in vitro

Seung Min Yoo, Ae Kyung Ahn, Taegun Seo, Hyo Bong Hong, Myung Ae Chung, Sang Don Jung, Haewol Cho, Myung Shin Lee

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

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.

Original languageEnglish
Pages (from-to)160-166
Number of pages7
JournalJournal of Virological Methods
Volume154
Issue number1-2
DOIs
StatePublished - Dec 2008

Keywords

  • Centrifugal enhancement
  • Human umbilical vein endothelial cells
  • Infection
  • Kaposi's sarcoma-associated herpesvirus

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