Hypomethylation of interleukin-4 and -6 promoters in T cells from systemic lupus erythematosus patients.
OBJECTIVE
DNA methylation regulates gene expression, and hypomethylation is associated with abnormal T-cell function in systemic lupus erythematosus (SLE). However, little is known about the methylation levels of the interleukin (IL)-4 and -6 promoters in SLE patients.


METHODS
T cells were isolated from 20 SLE patients and 10 healthy controls, activated in vitro in the presence or absence of 5- azacytidine (5-azaC), and their IL-4 and -6 transcripts were characterized using semiquantitative RT-PCR. Following bisulfate modification of their genomic DNA, the levels of DNA methylation in the IL-4 or -6 promoter were determined by nested PCR and direct sequencing.


RESULTS
The levels of IL-4 and -6 mRNA transcripts were significantly higher in SLE T cells, as compared with that in the controls. Furthermore, the treatment of healthy T cells with 5-azaC demethylated the CpG islands in the IL-4 or -6 promoter and increased IL-4 and -6 mRNA transcriptions. Importantly, the hypomethylation of the CpG islands in the IL-4 and -6 promoters displayed in SLE patients was similar to that of healthy T cells treated with 5-azaC. Finally, the hypomethylation levels of the CpG islands in the IL-4 and -6 promoters in lupus patients were significantly correlated to the IL-4 and -6 expressions.


CONCLUSIONS
The hypomethylation of the CpG islands of the IL-4 and -6 promoters accrued in T cells from SLE patients and was associated with the severity of SLE at the clinic.
Mi
Xiang-bin
XB
Department of Dermatology, Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Zeng
Fan-qin
FQ
D000328:Adult; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D001374:Azacitidine / Q000494:pharmacology; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D006801:Humans; D015847:Interleukin-4 / Q000235:genetics*; D015850:Interleukin-6 / Q000235:genetics*; D008180:Lupus Erythematosus, Systemic / Q000378:metabolism*; D008297:Male; D008745:Methylation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism*
D000964:Antimetabolites, Antineoplastic; D015850:Interleukin-6; D015847:Interleukin-4; D004247:DNA; D001374:Azacitidine