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학술지 Centrifugal Enhancement of Kaposi`s Sarcoma-associated Virus Infection of Human Endothelial Cells in Vitro
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유성민, 안애경, 서태건, 홍효봉, 정명애, 정상돈, 조해월, 이명신
Journal of Virological Methods, v.154 no.1-2, pp.160-166
08MC1600, 나노 입자를 이용한 고성능 바이오 센서 시스템, 정명애
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 제안 키워드
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