Asia Pac Allergy.  2015 Apr;5(2):114-122. 10.5415/apallergy.2015.5.2.114.

Reduced IRF7 response to rhinovirus unrelated with DNA methylation in peripheral mononuclear cells of adult asthmatics

• Kim SH
• Lim KH
• Park HK
• Lee SY
• Kim SH
• Kang HR
• Park HW
• Chang YS
• Cho SH

• KMID: 2397058
• DOI: http://doi.org/10.5415/apallergy.2015.5.2.114

Abstract

BACKGROUND
Human rhinoviruses are the major cause of asthma exacerbation in both children and adults. Recently, impaired antiviral interferon (IFN) response in asthmatics has been indicated as a primary reason of the susceptibility to respiratory virus, but the mechanism of defective IFN production is little understood to date. The expression of IFN regulatory factor 7 (IRF7), a transcriptional factor for virus-induced type I IFN production is known to be regulated epigenetically by DNA methylation.
OBJECTIVE
We aimed to investigate the expression of IFN-α, IFN-β, and IRF7 in response to rhinovirus infection in the adult asthmatics and evaluate DNA methylation status of IRF7 gene promotor.
METHODS
Twenty symptomatic adult asthmatics and 10 healthy subjects were enrolled and peripheral blood was collected from each subject. Peripheral blood mononuclear cells (PBMCs) were isolated, cultured, and ex vivo stimulated with rhinovirus-16. The mRNA expressions of IFN-α, IFN-β, and IRF7 were analyzed using real time quantitative polymerase chain reaction. Genomic DNA was isolated from untreated PBMCs and the methylation status of IRF7 gene promotor was investigated using bisulfite pyrosequencing.
RESULTS
The mean age of asthmatics was 45.4 ± 15.7 years and 40% was male, which were not different with those of control group. Asthmatics showed significantly decreased mRNA expressions (relative expression to beta-actin) of IFN-α and IFN-β compared with normal control. The mRNA expression of IRF7 in the asthmatics was also significantly lower than those in the normal control. No significant difference of DNA methylation was observed between asthmatics and controls in all analyzed positions of IRF7 promotor CpG loci.
CONCLUSION
The mRNA expression of type I IFN in response to rhinovirus was impaired in the PBMCs of adult asthmatics. The mRNA expression of IRF7, transcriptional factor inducing type I IFN was also reduced, but not caused by altered DNA methylation in the IRF7 gene promotor.

Keyword

Asthma; DNA methylation; Interferon type I; Rhinovirus

MeSH Terms

Adult*
Asthma
Child
DNA Methylation*
DNA*
Healthy Volunteers
Humans
Interferon Type I
Interferons
Male
Methylation
Polymerase Chain Reaction
Rhinovirus*
RNA, Messenger
DNA
Interferon Type I
Interferons
RNA, Messenger

Figure

• 

  Fig. 1 Expressions of type I interferon (IFN) and interferon regulatory factor 7 (IRF7) mRNA in response to rhinovirus stimulation in the peripheral blood mononuclear cells of asthmatics and healthy subjects. IFN-α (A), IFN-β (B), and IRF7 (C) values in panels A, B and C are mean ± standard error of the mean. RV 16, rhinovirus 16. *p < 0.05. **p < 0.01.

• 

  Fig. 2 Protein levels of type I interferon (IFN) in the supernatant from rhinovirus-stimulated peripheral blood mononuclear cells of asthmatics and healthy subjects. IFN-α (A) and IFN-β (B) values in panels A and B are mean ± standard error of the mean. RV 16, rhinovirus 16. *p < 0.05.

• 

  Fig. 3 DNA methylation analysis in the IRF7 gene promotor. (A) Analyzed CpG loci in the IRF7 gene promotor: underlined, analyzed region; black bold, CpG loci; red bold, positions evaluated finally. (B) Comparison of DNA methylation status of IRF7 gene promotor between asthmatics and healthy subjects.

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