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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

CD4 promoter methylation levels at 5 (A), 10 (B) and 21dpi (C). Pyrosequencing result of the promoter methylation level of CD4 gene before and after MDV infection at different time points. A decrease of promoter methylation level was observed only L72 chickens. 5dpi: 5 days post infection; 10dpi: 10 days post infection; 21dpi: 21 days post infection. L63.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken; L72.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken. n=4 for each line.
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Figure 1: CD4 promoter methylation levels at 5 (A), 10 (B) and 21dpi (C). Pyrosequencing result of the promoter methylation level of CD4 gene before and after MDV infection at different time points. A decrease of promoter methylation level was observed only L72 chickens. 5dpi: 5 days post infection; 10dpi: 10 days post infection; 21dpi: 21 days post infection. L63.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken; L72.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken. n=4 for each line.

Mentions: To determine the promoter methylation level of the CD4 gene, a DNA sequence containing the CpG islands from the CD4 gene promoter region (sequence shown in Table 1) was downloaded from UCSC (http://genome.ucsc.edu) and the methylation level was determined by pyrosequencing. The CpGs in the promoter of CD4 exhibits a high (>70%) methylation level in both L63 and L72 chickens before MDV infection. During MD progression, no significant methylation changes of CD4 promoter were detected in L63 chickens at 5, 10 and 21 dpi or in L72 chickens at 5 and 10 dpi (P>0.05, Figure 1, and Figure 1A and 1B); however, the significant down-regulation of CD4 promoter methylation level was observed at 21 dpi in L72 chickens (P< 0.05, Figure 1C). The result from the exact F test revealed that the CD4 promoter methylation pattern in L72 infected samples at 21 dpi was significantly different from any other groups (Figure 2).


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)

CD4 promoter methylation levels at 5 (A), 10 (B) and 21dpi (C). Pyrosequencing result of the promoter methylation level of CD4 gene before and after MDV infection at different time points. A decrease of promoter methylation level was observed only L72 chickens. 5dpi: 5 days post infection; 10dpi: 10 days post infection; 21dpi: 21 days post infection. L63.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken; L72.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken. n=4 for each line.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: CD4 promoter methylation levels at 5 (A), 10 (B) and 21dpi (C). Pyrosequencing result of the promoter methylation level of CD4 gene before and after MDV infection at different time points. A decrease of promoter methylation level was observed only L72 chickens. 5dpi: 5 days post infection; 10dpi: 10 days post infection; 21dpi: 21 days post infection. L63.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken; L72.Non: noninfected control of L63 chicken; L63.Inf: infected L63 chicken. n=4 for each line.
Mentions: To determine the promoter methylation level of the CD4 gene, a DNA sequence containing the CpG islands from the CD4 gene promoter region (sequence shown in Table 1) was downloaded from UCSC (http://genome.ucsc.edu) and the methylation level was determined by pyrosequencing. The CpGs in the promoter of CD4 exhibits a high (>70%) methylation level in both L63 and L72 chickens before MDV infection. During MD progression, no significant methylation changes of CD4 promoter were detected in L63 chickens at 5, 10 and 21 dpi or in L72 chickens at 5 and 10 dpi (P>0.05, Figure 1, and Figure 1A and 1B); however, the significant down-regulation of CD4 promoter methylation level was observed at 21 dpi in L72 chickens (P< 0.05, Figure 1C). The result from the exact F test revealed that the CD4 promoter methylation pattern in L72 infected samples at 21 dpi was significantly different from any other groups (Figure 2).

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