GATA3 siRNA inhibits the binding of NFAT1 to interleukin-13 promoter in human T cells.
BACKGROUND
Interleukin-13 (IL-13) is recognized to be a key modulator in the pathogenesis of Th2-induced allergic inflammation. Transcription factors GATA3 and NFAT1 have been both implicated in the regulation of Th2 cytokines. We previously demonstrated the GATA3-NFAT1 association during human T cell activation. However, the function of the GATA3-NFAT1 complex in Th2 cytokines regulation is still unknown. Small interference RNA (siRNA) was constructed to knock down GATA3 expression in Hut-78 cells to investigate the possible role of GATA3-NFAT1 complex in IL-13 transcription.


METHODS
Cells were stimulated with anti-CD3 plus anti-CD28 antibodies to mimic in vivo antigen-mediated co-stimulation; the expression of IL-13 mRNA was determined by real-time PCR; chromation immunoprecipitation (CHIP) assay was employed to investigate the NFAT1 binding to IL-13 promoter.


RESULTS
GATA3 siRNA suppressed the expression of GATA3 both in mRNA and protein levels in Hut-78 cells. The binding of NFAT1 to IL-13 promoter was inhibited by GATA3 siRNA in activated T cells, which was followed by the reduction of IL-13 transcription.


CONCLUSIONS
GATA3-NFAT1 complex may play an important role in the regulation of IL-13 transcription in human T cells.
Yao
Xin
X
Department of Respiratory Disease, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
Yang
Yan
Y
He
Hai-yan
HY
Wang
Min
M
Yin
Kai-sheng
KS
Huang
Mao
M
D002478:Cells, Cultured; D050990:GATA3 Transcription Factor / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D018793:Interleukin-13 / Q000235:genetics*; D050778:NFATC Transcription Factors / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D018793:Interleukin-13; D050778:NFATC Transcription Factors; C494006:NFATC2 protein, human; D034741:RNA, Small Interfering