The Effects Of The Interaction Between GATA3 And NFAT1 On IL-13 Gene Expression In Human T Cells

Asthma is a complex inflammatory disease of the lung characterized by airway hyperresponsiveness(AHR),eosinophilic inflammation and mucus hypersecretion.The inflammatory process,which is the basis of AHR and mucus hypersecretion,is believed to be a result of inappropriate immune responses to common aeroallergens in genetically susceptible individuals. It has been hypothesized that CD4⁺T cells that produce a Th2 pattern of cytokines(IL-4,IL-5,IL-13)play a pivotal role in the pathogenesis of asthma.However,the therapy targeted at IL-4 and IL-5 proved little efficacy.IL-13 has been recognized as a critical regulator of the allergic response.Blockade of IL-13 markedly inhibits allergen-induced airway eosinophilia,hyperresponsiveness,mucus production and airway remodeling.However,the molecular mechanisms regulating IL-13 production are still largely unknown.GATA3 and NFAT1 both play an important role in the expression of IL-13 gene.To investigate the interaction between GATA3 and NFAT1 on IL-13 gene expression,human T cell lines(Hut-78 cells)were stimulated with anti-CD3 and anti-CD28 antibodies to mimic antigen-mediated co-stimulation in vivo. PARTâ… :The synergistic interaction between NFAT1 and GATA3 On gene expression of IL-13 in human T cellsObjective:To investigate the synergistic interaction between NFAT1 and GATA3 on IL-13 gene expression in human T cells due to CD3/CD28 co-stimulation.Methods:Hut-78 cells were stimulated with anti-CD3/CD28 monoclonal antibodies at the concentration of 10μg/ml and 5μg/ml respectively.The interaction between NFAT1 and GATA3 in several times(0min,30min, 1h,2h)was detected by immunoprecipitation assays.Chromatin immunoprecipitation assays were used to investigate the binding of GATA3 to the promoter of IL-13.Results:Anti-CD3/CD28 co-stimulation induced the binding of GATA3 and NFAT1 in a time-dependant manner.FK-505 can inhibit the binding of GATA3 to NFAT1,and thus the binding of GATA3 to IL- 13 promoter.Conclusions:These results demonstrate the important role of the synergistic interaction between NFAT1 and GATA3 in the regulation of IL-13 gene expression in Hut-78 cells following CD3/CD28 co-stimulation. PARTâ…¡:The role of FP on the gene expression of IL-13 in actived human T cellsObjective:To study the effect of FP on the GATA3-NFAT1 interaction and subsequent GATA3 binding to IL-13 promoter in human T cells due to CD3/CD28 co-stimulation and investigate the anti-inflammatory mechanism of FP.Methods:Hut-78 cells were stimulated with anti-CD3/CD28 monoclonal antibodies at the concentration of 10μg/ml and 5μg/ml respectively.The effect of FP on the GATA3-NFAT1 interaction in 0min,30min,1h and 2h were detected by immunoprecipitation assays.Chromatin immunoprecipitation assays were used to investigate the binding of GATA3 to the promoter of IL- 13.Results:Anti-CD3/CD28 co-stimulation induced the binding of GATA3 and NFAT1 in a time-dependant manner,which was partially inhibited by FP at the concentration of 10^-8M FP also inhibited the binding of GATA3 to the promoter of IL- 13.Conclusions:FP can inhibit IL-13 gene expression in actived human T cells,which may be associated with inhibiting of the formation of GATA3-NFAT1 compound and the binding of GATA3 to the promoter of IL-13. PARTâ…¢:Arsenic trioxide inhibits the gene expression of IL-13 in activated T cellsObjective:To study the interleukin 13(IL-13)gene expression in T cells due to CD3/CD28 co-stimulation and the effect of arsenic trioxide(AT) on IL-13 gene expression.Methods:Hut-78 cells were stimulated with anti-CD3/CD28 monoclonal antibodies(10μg/ml and 5μg/ml,respectively).IL-13 mRNA level in Hut-78 cells was measured in groups with or without AT treatment.The effect of AT on the GATA3-NFAT1 compound in 0min,30min,1h and 2h were detected by immunoprecipitation assays.Results:The level of IL-13 mRNA increased in Hut-78 cells due to CD3/CD28 co-stimulation.However AT inhibited the gene expression of IL-13.AT inhibited the binding of GATA3 and NFAT1 in actived human T cells in 2h.Conclusions:AT would inhibit the gene expression of IL-13 in activated T cells,which might be partially through inhibiting the formation of the GATA3-NFAT1 compound.