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

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-12p35siRNA、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-10mRNA和IL-10的升高。结论 IL-12p35亚基特异性siRNA能够成功阻止IL-12mRNA的表达，降低了具有生物活性IL-12p70的浓度，提高IL-10的浓度。

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

Objective To explore biological characteristics of IL-12p35 siRNA and IL-12p40 siR- NA gene-modified DC in vitro.Methods The recombinant adenovirus expression plasmid (Ad-IL-12p35 siRNA,Ad-IL-12p40 siRNA,Ad-HKsiRNA) was transfected 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 Transfection of DC with siR- NA specific for the IL-12 p35 gene and IL-12p40 resulted in potent suppression of gene expression but blockade of bioactive IL-12 p70 production only in IL-12p35 gene-modified DC without affecting unrelat- ed genes.Inhibition of IL-12 was associated with the increased IL-10 production in IL-12p35 gene-modi- fied DC.Conclusion RNAi can be used for immune modulation by targeting DC gene expression.The IL-12p35 mRNA expression and bioactive IL-I2 p70 production was blockaded and the levels of IL-10 were increased by transfection of DC with siRNA specific for the IL-12 p35 gene.