Integration of Epstein-Barr virus in Burkitt's lymphoma cells leads to a region of enhanced chromosome instability

Jox, Andrea; Rohen, Corina; Belge, Gazanfer; Bartnitzke, Sabine; Pawlita, Michael; Diehl, Volker; Bullerdiek, Jörn; Wolf, Jürgen

doi:10.1093/annonc/8.suppl_2.s131

Cited by 31 publications

(26 citation statements)

References 16 publications

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“…Our results support this view, but also indicate that in latently and lytically infected cell lines, viral integration is a considerably more common phenomenon than previously assumed. 44,50 Moreover, the presence of small numbers of episomal and chromosomally integrated EBV copies was observed in cells of newly transformed mantle cell lymphoma cell lines (Drs ME Williams and DG Bebb, personal communication), supporting the view that EBV integration mechanisms may be initiated shortly after infection. Latent membrane protein 1 is generally regarded to be a marker of latent viral infection, 51 its expression was seen in all latently infected cell lines and was associated with EBV DNA signal types 1-3, confirming that these patterns were representative of latent infection.…”

Section: Discussionmentioning

confidence: 78%

Rapid determination of Epstein–Barr virus latent or lytic infection in single human cells using in situ hybridization

et al. 2004

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Epstein-Barr (EBV) virus is associated with malignancies such as lymphoma and carcinoma. Infection of cells with EBV may result in either lytic infection with production of viral particles, characterized by the presence of linear DNA forms, or latent infection, characterized by either episomal or integrated DNA forms. To examine whether the different lytic and latent EBV DNA forms can reliably be distinguished in single human cells, in situ hybridization was performed in EBV-positive cell lines. Immunocytochemistry and Southern blot analysis were performed supplementary to in situ hybridization. In latent infection, three in situ hybridization patterns were observed: large-disperse (episomal), small-punctate (integrated) and combined (both), signal types 1, 2 and 3 respectively. These were associated with expression of latent membrane protein 1, but not with Z fragment of Epstein-Barr replication activator or viral capsid antigen. In lytic infection, three additional in situ hybridization patterns were observed: nuclear membrane associated, bubble (filling up the nucleus) and spillover (covering the lysed cells) signals types 4, 5 and 6 respectively. Signal types 4 and 5 were associated with expression of latent membrane protein 1 and Z fragment of Epstein-Barr replication activator but not viral capsid antigen, whereas type 6 was associated with expression of viral capsid antigen only. Southern blot analysis confirmed these results; however, low copy numbers of integrated virus were often missed by Southern blot, confirming that in situ hybridization is more sensitive in determining the presence of all types of EBV DNA. In situ hybridization may prove useful in rapidly screening large series of tissue microarrays and other clinical specimens for the presence of lytic or latent EBV.

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Section: Discussionmentioning

confidence: 78%

Rapid determination of Epstein–Barr virus latent or lytic infection in single human cells using in situ hybridization

et al. 2004

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“…Both transmissible (Bernheim et al, 1981;Klein, 1986) and unstable chromosomal aberrations (Stollmann et al, 1985;Zattara-Cannoni et al, 1996;Chan et al, 2001) are present in EBV-associated malignancies, but the contribution of the virus to this phenotype is not understood. It was suggested that EBV might prime genomic instability by activating specific recombinases (Kuhn-Hallek et al, 1995) by integration (Jox et al, 1997) or by induction of oxidative stress (Gualandi et al, 2001), whereas latent viral genes such as LMP1 and LMP2 could contribute by modulating telomere function and DNA repair Liu et al, 2005), or by enhancing the sensitivity to DNAdamaging agents (O'Nions and Allday, 2003;Liu et al, 2004). However, there is no conclusive evidence for the mutagenic potential of the virus.…”

Section: Introductionmentioning

confidence: 99%

Epstein–Barr virus promotes genomic instability in Burkitt's lymphoma

et al. 2007

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Epstein-Barr virus (EBV) has been implicated in the pathogenesis of human malignancies but the mechanisms of oncogenesis remain largely unknown. Genomic instability and chromosomal aberrations are hallmarks of malignant transformation. We report that EBV carriage promotes genomic instability in Burkitt's lymphoma (BL). Cytogenetic analysis of EBVÀ and EBV þ BL lines and their sublines derived by EBV conversion or spontaneous loss of the viral genome revealed a significant increase in dicentric chromosomes, chromosome fragments and chromatid gaps in EBV-carrying cells. Expression of EBV latency I was sufficient for this effect, whereas a stronger effect was observed in cells expressing latency III. Telomere analysis by fluorescent in situ hybridization revealed an overall increase of telomere size and prevalence of telomere fusion and double strand-break fusion in dicentric chromosomes from EBV þ cells. Phosphorylated H2AX, a reporter of DNA damage and ongoing repair, was increased in virus-carrying cells in the absence of exogenous stimuli, whereas efficient activation of DNA repair was observed in both EBV þ and EBVÀ cells following treatment with etoposide. These findings point to induction of telomere dysfunction and DNA damage as important mechanisms for EBV oncogenesis.

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“…2 During latency, EBV DNA persists as an episome in the nuclei of host cells, or can be integrated into the cellular genome via the TR sequences. [3][4][5][6][7] Integration may initiate transformation events by causing chromosomal instability, [8][9][10] or may provide a cis-acting mechanism for altering gene regulation or structure. 11,12 Hence, if integration has an effect on viral or cellular gene expression, 5,13 it could be directly linked to the oncogenic properties of EBV, 14 mediating continuous cell proliferation.…”

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confidence: 99%

Visualization of episomal and integrated Epstein‐Barr virus DNA by fiber fluorescence in situ hybridization

Reisinger

Rumpler

Lion

et al. 2005

Intl Journal of Cancer

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For many Epstein-Barr virus (EBV)-associated malignancies, it is still a matter of controversy whether infected cells harbor episomal or chromosomally integrated EBV genomes or both. It is well established that the expression of EBV genes per se carries oncogenic potential, but the discrimination between episomal and integrated forms is of great relevance because integration events can contribute to the oncogenic properties of EBV, whereas host cells that exclusively harbor viral episomes may not carry the risks mediated by chromosomal integration. This notion prompted us to establish a reliable technique that not only allows to unequivocally discriminate episomal from integrated EBV DNA, but also provides detailed insights into the genomic organization of the virus. Here, we show that dynamic molecular combing of host cell DNA combined with fluorescence in situ hybridization (FISH) using EBV-specific DNA probes facilitate unambiguous discrimination of episomal from integrated viral DNA. Furthermore, the detection of highly elongated internal repeat 1 (IR1) sequences provides evidence that this method permits detection of major genomic alterations within the EBV genome. Thus, fiber FISH may also provide valuable insights into the genomic organization of viral genomes other than EBV. ' 2005 Wiley-Liss, Inc.Key words: EBV; fiber FISH; episomal; integration Epstein-Barr virus (EBV, human herpes virus 4) is associated with a number of benign and malignant diseases, including infectious mononucleosis, Burkitt's lymphoma, Hodgkin's disease, immunoblastic and transplantation-associated lymphomas. The virus preferentially infects B lymphocytes. 1 Upon infection, the linear viral DNA is covalently closed by its terminal repeat (TR) sequences and circularizes. 2 During latency, EBV DNA persists as an episome in the nuclei of host cells, or can be integrated into the cellular genome via the TR sequences. [3][4][5][6][7] Integration may initiate transformation events by causing chromosomal instability, [8][9][10] or may provide a cis-acting mechanism for altering gene regulation or structure. 11,12 Hence, if integration has an effect on viral or cellular gene expression, 5,13 it could be directly linked to the oncogenic properties of EBV, 14 mediating continuous cell proliferation. It has recently been shown that loss of expression of the putative tumor suppressor gene BACH2 in a Burkitt's lymphoma cell line is attributable to EBV integration within the first intron of the gene. 13 Another Burkitt's lymphoma cell line has been reported to contain integrated EBV DNA between the oncogenes REL and BCL11A associated with an increased expression of REL. 15 The notion that host cells, which exclusively contain episomal viral DNA, are not susceptible to the risks linked to chromosomal integration emphasizes the necessity of having a reliable method permitting distinction between episomal and integrated EBV DNA. We have therefore established a technique based on DNA fiber analysis, facilitating detailed insights into the genomic or...

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Integration of Epstein-Barr virus in Burkitt's lymphoma cells leads to a region of enhanced chromosome instability

Cited by 31 publications

References 16 publications

Rapid determination of Epstein–Barr virus latent or lytic infection in single human cells using in situ hybridization

Rapid determination of Epstein–Barr virus latent or lytic infection in single human cells using in situ hybridization

Epstein–Barr virus promotes genomic instability in Burkitt's lymphoma

Visualization of episomal and integrated Epstein‐Barr virus DNA by fiber fluorescence in situ hybridization

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