白细胞介素(IL)-12干扰RNA对树突状细胞IL-12 mRNA及细胞因子的影响

Effect of IL-12 siRNA on IL-12 mRNA and cytokines of dendritic cells

目的探讨白细胞介素(IL)-12干扰RNA(siRNA,IL-12p35 siRNA、IL-12p40 siRNA)重组腺病毒载体感染树突状细胞后对其功能的影响。方法用重组腺病毒载体(IL-12p35 siRNA、IL-12p40 siRNA、阴性对照HK siRNA)感染DC,检测IL-12p35亚基mRNA、IL-12p40亚基mRNA和IL-10 mRNA的表达,同时检测上清中IL-12p70、IL-10水平。结果IL-12p35 siRNA、IL-12p40 siRNA重组腺病毒载体分别感染树突状细胞后,干扰了IL-12p35亚基mRNA和IL-12p40亚基mRNA的表达,但是只有IL-12p35 siRNA重组腺病毒载体感染DC上清中IL-12p70明显减少,同时引起了IL-10 mRNA和IL-10的升高。结论IL-12p35亚基特异性siRNA能够成功阻止IL-12 mRNA的表达,降低了具有生物活性IL-12p70的浓度,提高IL-10的浓度。

Objective To explore biological characteristics of IL-12p35 siRNA and IL-12p40 siRNA gene-modified DC in vitro. Methods The recombinant adenovirus expression plasmid （Ad-IL-12p35 siRNA,Ad-IL-12p40 siRNA, Ad-HKsiRNA） was transfeeted into bone marrow-derived dendritic cells （BMDC）. IL-12p35 mRNA, IL-12p40 mRNA and IL-10 mRNA were detected by RT-PCR. IL-12p70 and IL-10 in culture supernatant were examined by ELISA assay. Results Transfeetion of DC with siRNA specific for the IL-12 p35 gene and IL-12p40 resulted in potent suppression of gene expression but blockade of bioaetive IL-12 p70 production only in IL-12p35 gene-modified DC without affecting unrelated genes. Inhibition of IL-12 was associated with the increased IL-10 production in IL-12p35 gene-modified DC. Conclusion RNAi can be used for immune modulation by targeting DC gene expression. The IL-12p35 mRNA expression and bioaetive IL-12 p70 production was blockaded and the levels of IL-10 were increased by transfeetion of DC with siRNA specific for the IL-12 p35 gene.