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Down-regulation of promoter methylation level of CD4 gene after MDV infection in MD-susceptible chicken line.

Luo J, Yu Y, Zhang H, Tian F, Chang S, Cheng HH, Song J - BMC Proc (2011)

Bottom Line: Chickens from each line were divided into two groups with one group infected by MDV and the other group as uninfected controls.Then, promoter DNA methylation levels of the CD4 gene were measured by Pyrosequencing; and gene expression analysis was performed by quantitative PCR.The methylation down-regulation of the CD4 promoter is negatively correlated with up-regulation of CD4 gene expression in the L72 spleen at 21 dpi.

View Article: PubMed Central - HTML - PubMed

Affiliation: Animal & Avian Sciences Department, University of Maryland, College Park, Maryland, 20740, USA. songj88@umd.edu.

ABSTRACT

Background: Marek's disease virus (MDV) is an oncovirus that induces lymphoid tumors in susceptible chickens, and may affect the epigenetic stability of the CD4 gene. The purpose of this study was to find the effect of MDV infection on DNA methylation status of the CD4 gene differed between MD-resistant (L63) and -susceptible (L72) chicken lines.

Methods: Chickens from each line were divided into two groups with one group infected by MDV and the other group as uninfected controls. Then, promoter DNA methylation levels of the CD4 gene were measured by Pyrosequencing; and gene expression analysis was performed by quantitative PCR.

Results: Promoter methylation of the CD4 gene was found to be down-regulated in L72 chickens only after MDV infection. The methylation down-regulation of the CD4 promoter is negatively correlated with up-regulation of CD4 gene expression in the L72 spleen at 21 dpi.

Conclusions: The methylation fluctuation and mRNA expression change of CD4 gene induced by MDV infection suggested a unique epigenetic mechanism existed in MD-susceptible chickens.

No MeSH data available.


Related in: MedlinePlus

Relative mRNA expression of CD4 gene at 21dpi. Real-time quantitative PCR was used to detect the mRNA expression level of CD4 gene in different chicken lines with or without MDV infection at 21dpi. The relative expression level of CD4 gene was normalized to a house keeping gene GAPDH. Non.: noninfected control samples; Inf.: infected samples. n=4 for each line and treatment.
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Figure 3: Relative mRNA expression of CD4 gene at 21dpi. Real-time quantitative PCR was used to detect the mRNA expression level of CD4 gene in different chicken lines with or without MDV infection at 21dpi. The relative expression level of CD4 gene was normalized to a house keeping gene GAPDH. Non.: noninfected control samples; Inf.: infected samples. n=4 for each line and treatment.

Mentions: To ascertain if the CD4 gene transcription level is influenced by its promoter methylation changes at 21 dpi, we conducted quantitative PCR. We found a significantly higher expression of CD4 gene in L72 infected samples compared with noninfected control samples (P<0.05) (Figure 3), whereas no significant up or down-regulation of CD4 expression was detected in L63 chickens after MDV infection (P>0.05). Hereinafter, further correlation analysis showed that methylation level of all the detected CpG sites existed a negatively relationship with CD4 gene expression in L72 chicken at 21dpi (Figure 4).


Down-regulation of promoter methylation level of CD4 gene after MDV infection in MD-susceptible chicken line.

Luo J, Yu Y, Zhang H, Tian F, Chang S, Cheng HH, Song J - BMC Proc (2011)

Relative mRNA expression of CD4 gene at 21dpi. Real-time quantitative PCR was used to detect the mRNA expression level of CD4 gene in different chicken lines with or without MDV infection at 21dpi. The relative expression level of CD4 gene was normalized to a house keeping gene GAPDH. Non.: noninfected control samples; Inf.: infected samples. n=4 for each line and treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108237&req=5

Figure 3: Relative mRNA expression of CD4 gene at 21dpi. Real-time quantitative PCR was used to detect the mRNA expression level of CD4 gene in different chicken lines with or without MDV infection at 21dpi. The relative expression level of CD4 gene was normalized to a house keeping gene GAPDH. Non.: noninfected control samples; Inf.: infected samples. n=4 for each line and treatment.
Mentions: To ascertain if the CD4 gene transcription level is influenced by its promoter methylation changes at 21 dpi, we conducted quantitative PCR. We found a significantly higher expression of CD4 gene in L72 infected samples compared with noninfected control samples (P<0.05) (Figure 3), whereas no significant up or down-regulation of CD4 expression was detected in L63 chickens after MDV infection (P>0.05). Hereinafter, further correlation analysis showed that methylation level of all the detected CpG sites existed a negatively relationship with CD4 gene expression in L72 chicken at 21dpi (Figure 4).

Bottom Line: Chickens from each line were divided into two groups with one group infected by MDV and the other group as uninfected controls.Then, promoter DNA methylation levels of the CD4 gene were measured by Pyrosequencing; and gene expression analysis was performed by quantitative PCR.The methylation down-regulation of the CD4 promoter is negatively correlated with up-regulation of CD4 gene expression in the L72 spleen at 21 dpi.

View Article: PubMed Central - HTML - PubMed

Affiliation: Animal & Avian Sciences Department, University of Maryland, College Park, Maryland, 20740, USA. songj88@umd.edu.

ABSTRACT

Background: Marek's disease virus (MDV) is an oncovirus that induces lymphoid tumors in susceptible chickens, and may affect the epigenetic stability of the CD4 gene. The purpose of this study was to find the effect of MDV infection on DNA methylation status of the CD4 gene differed between MD-resistant (L63) and -susceptible (L72) chicken lines.

Methods: Chickens from each line were divided into two groups with one group infected by MDV and the other group as uninfected controls. Then, promoter DNA methylation levels of the CD4 gene were measured by Pyrosequencing; and gene expression analysis was performed by quantitative PCR.

Results: Promoter methylation of the CD4 gene was found to be down-regulated in L72 chickens only after MDV infection. The methylation down-regulation of the CD4 promoter is negatively correlated with up-regulation of CD4 gene expression in the L72 spleen at 21 dpi.

Conclusions: The methylation fluctuation and mRNA expression change of CD4 gene induced by MDV infection suggested a unique epigenetic mechanism existed in MD-susceptible chickens.

No MeSH data available.


Related in: MedlinePlus