title: Centrifugal enhancement of Kaposi's sarcoma-associated virus infection of human endothelial cells in vitro.
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.
authors.affiliation: Department of Microbiology and Immunology, Eulji University School of Medicine, Daejeon, South Korea.
authors.forename: Seung-Min
authors.initials: SM
authors.lastname: Yoo
authors.forename: Ae-Kyung
authors.initials: AK
authors.lastname: Ahn
authors.forename: Taegun
authors.initials: T
authors.lastname: Seo
authors.forename: Hyo Bong
authors.initials: HB
authors.lastname: Hong
authors.forename: Myung-Ae
authors.initials: MA
authors.lastname: Chung
authors.forename: Sang-Don
authors.initials: SD
authors.lastname: Jung
authors.forename: Haewol
authors.initials: H
authors.lastname: Cho
authors.forename: Myung-Shin
authors.initials: MS
authors.lastname: Lee