Submitted by Rona S Scott (Louisiana State University Health Sciences Center-Shreveport, USA) on Jun 30 2014
Platform: microarray – [HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array
Pubmed: 25056883
Summary The oral cavity is the persistent reservoir for EBV with lifelong infection of resident epithelial and B cells. Infection of these cell types results in distinct EBV gene expression patterns that are regulated by epigenetic modifications involving DNA methylation and chromatin structure. Such regulation of EBV gene expression relies on viral manipulation of the host epigenetic machinery that may inadvertently result in long-lasting, oncogenic host epigenetic reprogramming. To test this hypothesis in the context of EBV infection of epithelial cells, we established a transient infection model to identify the epigenetic consequences after EBV infection of immortalized normal oral keratinocytes and subsequent viral loss.
With mounting evidence that EBV can induce epigenetic alterations, we developed a transient infection model where a clonal derivative (designated cl1) from a human telomerase immortalized normal oral keratinocyte (NOK) cell line was infected with a recombinant Akata EBV carrying neomycin resistance and GfP cassette in place of the BXLF1 open reading frame. Infected cells were passaged ten times, and then selection pressure was removed for an additional ten passages to allow for viral loss. Three transiently-infected EBV-negative clones were identified by single cell cloning (designated cl1, cl3, cl4). Uninfected parental clone and cells transfected with PTRUF5 plasmid were passaged alongside the transiently-infected clones. The transcription profiles were analyzed using Affymetrix microarray U133Plus2.0 arrays in duplicate.
ID | Title | Cell Type | Timepoint | Reported Virus | Virus Species | Exclusion Reason |
---|---|---|---|---|---|---|
GSM1422231 | NOK cl1 uninfected p31 | keratinocyte cell line | uninfected | Uninfected | ||
GSM1422232 | NOK cl1 uninfected p32 | keratinocyte cell line | uninfected | Uninfected | ||
GSM1422233 | NOK cl1 pTURF transfected p23 | keratinocyte cell line | vector transfected | Uninfected | ||
GSM1422234 | NOK cl1 pTURF transfected p24 | keratinocyte cell line | vector transfected | Uninfected | ||
GSM1422235 | NOK cl1 AkataBX-1 cl4 p31 EBV+ | keratinocyte cell line | EBV | Human gammaherpesvirus 4 | ||
GSM1422236 | NOK cl1 AkataBX-1 cl4 p32 EBV+ | keratinocyte cell line | EBV | Human gammaherpesvirus 4 | ||
GSM1422237 | NOK cl1 AkataBX-1 cl1 p29 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |
|
GSM1422238 | NOK cl1 AkataBX-1 cl1 p30 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |
|
GSM1422239 | NOK cl1 AkataBX-1 cl3 p29 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |
|
GSM1422240 | NOK cl1 AkataBX-1 cl3 p30 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |
|
GSM1422241 | NOK cl1 AkataBX-1 cl4 p29 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |
|
GSM1422242 | NOK cl1 AkataBX-1 cl4 p30 EBV- | keratinocyte cell line | EBV(-) | Uninfected | Extra interventions transiently infected |