Quantitative profiling of housekeeping and Epstein-Barr virus gene transcription in Burkitt lymphoma cell lines using an oligonucleotide microarray

Bernasconi, Michele; Berger, Christoph; Sigrist, Jürg A.; Bonanomi, Athos; Sobek, Jens; Niggli, Felix; Nadal, David

doi:10.1186/1743-422x-3-43

Cited by 31 publications

(11 citation statements)

References 34 publications

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“…The latency of EBV-associated malignancies is classified into three groups according to the EBV gene expression pattern; these latency patterns are used to classify the types of EBV-associated cancers (Fukayama et al , 1998). Namalwa, known as latency I/II, expresses EBNA1 and EBNA2, but not LMP1 and LMP2 (Bernasconi et al , 2006; Satoh et al , 2002). Also, we did not detect LMP2A expression in Namalwa cells by Western blot analysis, but did detect low levels of LMP2A mRNA by RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

Characterization of naturally Epstein–Barr virus-infected gastric carcinoma cell line YCCEL1

Kim

Seo

Choi

et al. 2013

Journal of General Virology

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Epstein-Barr virus (EBV) is a herpesvirus associated with lymphomas and carcinomas. While EBV-associated epithelial cell lines are good model systems to investigate the role of EBV in carcinoma, only a few cell lines are available as they are hard to acquire. A greater variety of naturally EBV-infected cell lines which are derived from tumour patients are needed to represent various features of EBVaGC. We characterized cell line YCCEL1, established from a Korean EBVaGC patient, to ascertain whether it can be used to study the roles of EBV in EBVaGC. The expression of EBV genes and cell surface markers was examined by in situ hybridization, RT-PCR, Western blot analysis, immunofluorescence assay and Northern blot analysis. EBV episomal status was analysed by Southern blotting and real-time PCR. This cell line expressed EBV nuclear antigen 1 (EBNA1) and latent membrane protein 2A (LMP2A), but not EBNA2, LMP2B nor LMP1. The majority of the lytic proteins were not detected in YCCEL1 cells either before or after treatment with 12-O-tetradecanoylphorbol-13-acetate. YCCEL1 cells expressed BART microRNAs (miRNAs) at high level but did not express BHRF1 miRNAs. YCCEL1 cells expressed cytokeratin, but not CD21 and CD19, suggesting CD21-independent EBV infection. The latent EBV gene and EBV miRNA expression pattern of YCCEL1 cells closely resembled that of general EBVaGC cases. Our results support the value of YCCEL1 cells as a good model system to study the role of EBV in gastric carcinogenesis.

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Section: Discussionmentioning

confidence: 99%

Characterization of naturally Epstein–Barr virus-infected gastric carcinoma cell line YCCEL1

Kim

Seo

Choi

et al. 2013

Journal of General Virology

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“…These were triplicate spots of b-actin, GAPDH, and eukaryotic elongation factor 1 a 1 (EEF1a1). Since viruses lack an appropriate control or Table 1 Primers used for validation of microarray data analysis a Forward primer is located within meq and reverse primer within R-LORF5 ''housekeeping'' gene to be used in statistical analysis, using host-encoded housekeeping genes for normalization of viral gene expression is a common practice in microarray studies [39][40][41][42][43][44][45][46][47]. The data was normalized by the publicly available software R v.2.4.1 using gcrma [48] followed by linear model analysis to identify differentially expressed genes between 5 and 15 dpi.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional profiling of Marek’s disease virus genes during cytolytic and latent infection

et al. 2008

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Marek's disease (MD), a lymphoproliferative disease of chicken is caused by a highly cell-associated alpha-herpesvirus, Marek's disease virus (MDV). MDV replicates in chicken lymphocytes and establishes a latent infection within CD4(+) T cells. The expression analysis of limited viral transcripts have revealed differences in gene expression pattern during cytolytic and latent phases of MDV infection. In this study, we conducted a global gene expression profiling of MDV using oligonucleotide-based Affymetrix GeneChip Chicken Genome Arrays. These arrays contain probe for more than 32,000 chicken transcripts and most of the known MDV genes and open reading frames. Two-week-old MD-susceptible chickens were inoculated with an oncogenic strain of MDV, and spleen samples were collected 5 and 15 days post inoculation (cytolytic and latent infection, respectively) for RNA isolation and microarray analysis. Array results displayed a significant differential pattern of viral transcriptome between the two phases of MDV infection. The expression levels of more than 78 MDV genes were increased during the cytolytic infection when compared to latent infection (2-11-fold increase). A 23-KD nuclear protein, meq oncoprotein, and R-LORF5 were among the few viral genes that were expressed during both phases of infection. In addition, there were at least 11 known and hypothetical genes that had no significant transcriptional activities during either stages of infection. These chicken genome arrays have considerable promise, as a valuable tool in understanding the molecular mechanism regulating MDV cytolytic and latent infection, and providing insights into the chicken gene expression pattern and associated biological pathways in response to different phases of viral pathogenesis.

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“…Gene expression was determined by quantitative reverse transcription polymerase chain reaction (RT-qPCR) using specific primers and probes for the housekeeping gene HMBS , the non-coding EBV encoded RNA EBER1 , the latency associated EBV genes EBNA1 , EBNA2 , LMP1 and LMP2A and for the two genes related to the lytic replication cycle of EBV, BZLF1 and BXLF2 , as described earlier [ 53 – 55 ]. Additionally, to detect BZLF1 gene expression in rM81 infected cells we used a different forward primer for the BZLF1 (5’-CAC GAC GTA CAA GGA AAC-3’) and LMP1 (5’-TGG AGG CCT TGG TCT ACT CCT-3’) primer/probe set, which we termed aBZLF1 and aLMP1 , respectively, due to the sequence homology to the Akata EBV strain.…”

Section: Methodsmentioning

confidence: 99%

Telomerase Activity Impacts on Epstein-Barr Virus Infection of AGS Cells

et al. 2015

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The Epstein-Barr virus (EBV) is transmitted from host-to-host via saliva and is associated with epithelial malignancies including nasopharyngeal carcinoma (NPC) and some forms of gastric carcinoma (GC). Nevertheless, EBV does not transform epithelial cells in vitro where it is rapidly lost from infected primary epithelial cells or epithelial tumor cells. Long-term infection by EBV, however, can be established in hTERT-immortalized nasopharyngeal epithelial cells. Here, we hypothesized that increased telomerase activity in epithelial cells enhances their susceptibility to infection by EBV. Using HONE-1, AGS and HEK293 cells we generated epithelial model cell lines with increased or suppressed telomerase activity by stable ectopic expression of hTERT or of a catalytically inactive, dominant negative hTERT mutant. Infection experiments with recombinant prototypic EBV (rB95.8), recombinant NPC EBV (rM81) with increased epithelial cell tropism compared to B95.8, or recombinant B95.8 EBV with BZLF1-knockout that is not able to undergo lytic replication, revealed that infection frequencies positively correlate with telomerase activity in AGS cells but also partly depend on the cellular background. AGS cells with increased telomerase activity showed increased expression mainly of latent EBV genes, suggesting that increased telomerase activity directly acts on the EBV infection of epithelial cells by facilitating latent EBV gene expression early upon virus inoculation. Thus, our results indicate that infection of epithelial cells by EBV is a very selective process involving, among others, telomerase activity and cellular background to allow for optimized host-to-host transmission via saliva.

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Quantitative profiling of housekeeping and Epstein-Barr virus gene transcription in Burkitt lymphoma cell lines using an oligonucleotide microarray

Abstract: Our results suggest uniform EBNA1 transcription levels in BL and that microarray profiling can reveal novel insights on quantitative EBV gene transcription and its impact on lymphocyte biology.

Cited by 31 publications

References 34 publications

Characterization of naturally Epstein–Barr virus-infected gastric carcinoma cell line YCCEL1

Characterization of naturally Epstein–Barr virus-infected gastric carcinoma cell line YCCEL1

Transcriptional profiling of Marek’s disease virus genes during cytolytic and latent infection

Telomerase Activity Impacts on Epstein-Barr Virus Infection of AGS Cells

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