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

Yoo, Seung‐Min; Ahn, Ae-Kyung; Seo, Taegun; Hong, Hyunsuk; Chung, Myung-Ae; Jung, Sang-Don; Cho, Haewol; Lee, Myung‐Shin

doi:10.1016/j.jviromet.2008.07.026

Cited by 32 publications

(29 citation statements)

References 33 publications

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“…The combined use of exogenous RTA expression and sodium butyrate treatment was shown to synergistically activate the lytic cycle, yielding substantial quantities of rKSHV.219 from Vero cells (68). Furthermore, the use of spin infection is capable of increasing the infection levels approximately up to 70-fold (68,73). Recently, the production of infectious cell-free KSHV was further refined by Myoung et al, who reported the utility of newly generated doxycycline-inducible RTA cell lines (called iVero and iSLK) for production of high-titer virus stocks, including BAC-derived stocks (49).…”

Section: Resultsmentioning

confidence: 99%

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Brulois

Chang

Lee

et al. 2012

J Virol

315

455

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Efficient genetic modification of herpesviruses such as Kaposi's sarcoma-associated herpesvirus (KSHV) has come to rely on bacterial artificial chromosome (BAC) technology. In order to facilitate this approach, we generated a new KSHV BAC clone, called BAC16, derived from the rKSHV.219 virus, which stems from KSHV and Epstein-Barr virus-coinfected JSC1 primary effusion lymphoma (PEL) cells. Restriction enzyme and complete sequencing data demonstrate that the KSHV of JSC1 PEL cells showed a minimal level of sequence variation across the entire viral genome compared to the complete genomic sequence of other KSHV strains. BAC16 not only stably propagated in bothEscherichia coliand mammalian cells without apparent genetic rearrangements, but also was capable of robustly producing infectious virions (∼5 × 107/ml). We also demonstrated the utility of BAC16 by generating deletion mutants of either the K3 or K5 genes, whose products are E3 ligases of the membrane-associated RING-CH (MARCH) family. While previous studies have shown that individual expression of either K3 or K5 results in efficient downregulation of the surface expression of major histocompatibility complex class I (MHC-I) molecules, we found that K5, but not K3, was the primary factor critical for the downregulation of MHC-I surface expression during KSHV lytic reactivation or followingde novoinfection. The data presented here demonstrate the utility of BAC16 for the generation and characterization of KSHV knockout and mutant recombinants and further emphasize the importance of functional analysis of viral genes in the context of the KSHV genome besides the study of individual gene expression.

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Section: Resultsmentioning

confidence: 99%

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Brulois

Chang

Lee

et al. 2012

J Virol

315

455

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“…Virus was resuspended in tonsil medium, and aliquots were stored at -80°C until use. Infectious units (IU) of recombinant virus stocks and various culture supernatants were determined on QBI293A cells by spinoculation, as described before (18,60). Briefly, serially diluted virus solution was infected on QBI293A cells by low-speed centrifugation (950 g for 90 minutes).…”

Section: Methodsmentioning

confidence: 99%

Active lytic infection of human primary tonsillar B cells by KSHV and its noncytolytic control by activated CD4+ T cells

Myoung¹,

Ganem²

2011

J. Clin. Invest.

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“…The isolated virus was used to infect cells as described previously [18] . Infection was conducted by centrifugation at 2,500 g for 60 min with 5 g ml -1 of polybrene (Sigma Aldrich).…”

Section: Preparation and Infection Of Kshv Bac36mentioning

confidence: 99%

“…Reverse transcription and quantitative PCR were conducted as described previously [18] . All samples, including a non-template control and internal glyceraldehyde 3-phosphate dehydrogenase (GAPDH) amplification controls, were examined in triplicate for each primer pair.…”

Section: Real-time Rt-pcrmentioning

confidence: 99%

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Expression of DcR3 and Its Effects in Kaposi’s Sarcoma-Associated Herpesvirus-Infected Human Endothelial Cells

Yoo

Jang²,

Kim³

et al. 2011

Intervirology

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Objective: Kaposi’s sarcoma-associated herpesvirus (KSHV) is classified as a gamma-herpesvirus and it causes Kaposi’s sarcoma in patients infected with the human immunodeficiency virus (HIV). Decoy receptor 3 (DcR3) is known as a decoy receptor for Fas ligand, LIGHT and TL1A and it can neutralize the biological effect of TL1A by inhibiting the TL1A-DR3 interaction in human endothelial cells. The present study examined the expression of DcR3 in human endothelial cells and its effect during the early stages of KSHV infection. Methods: The expression of DcR3 was assessed using real-time RT-PCR and ELISA in human umbilical cord vein endothelial cells (HUVECs) infected with KSHV. Cell proliferation and apoptosis of KSHV-infected HUVECs were assessed after treatment of infected cells with an anti-DcR3 antibody or recombinant human TL1A. Results: DcR3 expression was induced during the early phase of KSHV infection. Inhibition of DcR3 with anti-DcR3 antibodies or recombinant human TL1A-induced apoptosis in KSHV-infected HUVECs. Conclusion: The expression of DcR3 plays an important role in the prevention of apoptosis in HUVECs during the early phases of KSHV infection, thus ensuring the successful establishment and maintenance of the viral infection.

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

Cited by 32 publications

References 33 publications

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Construction and Manipulation of a New Kaposi's Sarcoma-Associated Herpesvirus Bacterial Artificial Chromosome Clone

Active lytic infection of human primary tonsillar B cells by KSHV and its noncytolytic control by activated CD4+ T cells

Expression of DcR3 and Its Effects in Kaposi’s Sarcoma-Associated Herpesvirus-Infected Human Endothelial Cells

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