Infection of Lymphoblastoid Cell Lines by Kaposi's Sarcoma-Associated Herpesvirus: Critical Role of Cell-Associated Virus

Myoung, Jinjong; Ganem, Don

doi:10.1128/jvi.05136-11

Cited by 55 publications

(61 citation statements)

References 49 publications

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“…Generation of iSLK cells carrying the ⌬ORF57 KSHV bacmid. In order to study the role of ORF57 in KSHV lytic-phase gene regulation, we used an epithelial cell line known as iSLK, which inducibly expresses the Rta (ORF50) protein only in the presence of doxycycline (37). iSLK cells exhibit robust KSHV lytic replication and infectious virion release when treated with doxycycline.…”

Section: Resultsmentioning

confidence: 99%

Identification of the Physiological Gene Targets of the Essential Lytic Replicative Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein

Verma

Krueger

et al. 2015

J Virol

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The Kaposi's sarcoma-associated herpesvirus (KSHV) ORF57 gene product is essential for lytic KSHV replication and virion production. Recombinant ORF57-null mutants fail to accumulate several lytic cycle mRNAs at wild-type levels, leading to decreased production of lytic proteins necessary for efficient replication. Several mechanisms by which ORF57 may enhance expression of lytic KSHV mRNAs have been proposed, including mRNA stabilization, mRNA nuclear export, increased polyadenylation, and transcriptional activation. ORF57 activity is also gene specific, with some genes being highly dependent on ORF57, whereas others are relatively independent. Most experiments have utilized transfection models for ORF57 and have not systematically examined the gene specificity and potential mechanisms of action of ORF57 in the context of KSHV-infected cells. In this study, the KSHV genes that are most highly upregulated by ORF57 during KSHV lytic replication were identified by a combination of high-throughput deep RNA sequencing, quantitative PCR, Northern blotting, and rapid amplification of cDNA ends methods. Comparison of gene expression from a ⌬ORF57 KSHV recombinant, a rescued ⌬ORF57 KSHV recombinant, and wild-type KSHV revealed that two clusters of lytic genes are most highly dependent on ORF57 for efficient expression. Despite contiguous location in the genome and shared polyadenylation of several of the ORF57-dependent genes, ORF57 regulation was promoter and polyadenylation signal independent, suggesting that the mRNAs are stabilized by ORF57. The eight genes identified to critically require ORF57 belong to both early and late lytic temporal classes, and seven are involved in DNA replication, virion assembly, or viral infectivity, explaining the essential role of ORF57 in infectious KSHV production. IMPORTANCEKaposi's sarcoma-associated herpesvirus (KSHV) is a human herpesvirus involved in the causation of several human cancers. The KSHV ORF57 protein is required for KSHV to replicate and produce infectious virus. We have identified several KSHV genes whose expression is highly dependent on ORF57 and shown that ORF57 increases expression of these genes specifically. These genes code for proteins that are required for the virus to replicate its DNA and to infect other cells. Identifying the targets and mechanism of action of ORF57 provides further approaches to discover antiviral therapy. K aposi's sarcoma associated herpesvirus (KSHV; human herpesvirus 8) is a human gammaherpesvirus associated with the development of Kaposi's sarcoma and several pathogenic lymphoproliferative syndromes (for a review, see reference 1). Lytic KSHV replication plays a role in pathogenesis by maintaining the infected cell reservoir, and lytic-phase KSHV proteins contribute to cell proliferation, inflammation, and immune evasion (2, 3). Understanding the mechanism of action of proteins required for lytic herpesvirus replication is therefore critical for further understanding of oncogenic mechanisms and the development of new antivi...

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Section: Resultsmentioning

confidence: 99%

Identification of the Physiological Gene Targets of the Essential Lytic Replicative Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein

Verma

Krueger

et al. 2015

J Virol

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“…The MC116 human B cell line provides a comparable system for KSHV. Its susceptibility to infection by cell-free virus overcomes the infection resistance of other previously analyzed B cell lines (30,32,33) as well as the requirement for the use of cell-associated virus, which is employed for infectivity studies of primary B cells (35) and stable B cell lines (58).…”

Section: Discussionmentioning

confidence: 99%

“…The Louckes human B cell line was shown to be susceptible to KSHV virions produced from KS lesions but, oddly, not KSHV virions from PEL-derived cell lines (34). The failure of B lymphoblastoid cell lines to support infection by cell-free KSHV virions could be bypassed via transmission of cell-associated virus from a stably infected cell line (35). Thus, a culture system involving a human continuous B cell line that is permissive for infection with experimentally produced cell-free KSHV would be of great value in the study of the various steps of the viral infection cycle.…”

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confidence: 99%

Efficient Infection of a Human B Cell Line with Cell-Free Kaposi's Sarcoma-Associated Herpesvirus

Dollery

Santiago-Crespo

Kardava

et al. 2014

J Virol

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“…Introduction of the viral DNA via electroporation into a B cell line resulted in latent persistence of KSHV (12). Only recently, KSHV-and EBV-negative BJAB cells were successfully infected with KSHV via cell-mediated transmission (13). Latently infected cells could be generated under selection (13,14) but were poorly permissive to lytic/productive replication after stimulation with valproate, sodium butyrate, or 12-O-tetradecanoyl-phorbol-13-acetate (TPA) (13).…”

mentioning

confidence: 99%

“…Only recently, KSHV-and EBV-negative BJAB cells were successfully infected with KSHV via cell-mediated transmission (13). Latently infected cells could be generated under selection (13,14) but were poorly permissive to lytic/productive replication after stimulation with valproate, sodium butyrate, or 12-O-tetradecanoyl-phorbol-13-acetate (TPA) (13). Most of our knowledge on the lytic/productive replication of KSHV in B cells stems from work on PEL cell lines.…”

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confidence: 99%

Activation of the B Cell Antigen Receptor Triggers Reactivation of Latent Kaposi's Sarcoma-Associated Herpesvirus in B Cells

Kati

Tsao

Guenther

et al. 2013

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV) is an oncogenic herpesvirus and the cause of Kaposi's sarcoma, primary effusion lymphoma (PEL) and multicentric Castleman's disease. Latently infected B cells are the main reservoir of this virus in vivo, but the nature of the stimuli that lead to its reactivation in B cells is only partially understood. We established stable BJAB cell lines harboring latent KSHV by cell-free infection with recombinant virus carrying a puromycin resistance marker. Our latently infected B cell lines, termed BrK.219, can be reactivated by triggering the B cell receptor (BCR) with antibodies to surface IgM, a stimulus imitating antigen recognition. Using this B cell model system we studied the mechanisms that mediate the reactivation of KSHV in B cells following the stimulation of the BCR and could identify phosphatidylinositol 3-kinase (PI3K) and X-box binding protein 1 (XBP-1) as proteins that play an important role in the BCR-mediated reactivation of latent KSHV.

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Infection of Lymphoblastoid Cell Lines by Kaposi's Sarcoma-Associated Herpesvirus: Critical Role of Cell-Associated Virus

Cited by 55 publications

References 49 publications

Identification of the Physiological Gene Targets of the Essential Lytic Replicative Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein

Identification of the Physiological Gene Targets of the Essential Lytic Replicative Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein

Efficient Infection of a Human B Cell Line with Cell-Free Kaposi's Sarcoma-Associated Herpesvirus

Activation of the B Cell Antigen Receptor Triggers Reactivation of Latent Kaposi's Sarcoma-Associated Herpesvirus in B Cells

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