CD80 and CD86 knockdown in dendritic cells regulates Th1/Th2 cytokine production in asthmatic mice
CD80 and CD86 knockdown in dendritic cells regulates Th1/Th2 cytokine production in asthmatic mice
复制标题
DOI：
10.3892/etm.2016.2989
复制
发表时间：
2016-03-01
影响因子：
2.7
通讯作者：
Ran, Pi-Xin
中科院分区：
文献类型：
--
作者：
Li, Jian-Guo;Du, Yu-Mo;Ran, Pi-Xin
树突状细胞(DC)与辅助性T细胞(Th)的活化和分化有关。分化簇(CD)80和CD86是DC中高表达的共刺激分子，在促进Th细胞向Th2细胞分化方面具有显著作用。然而，CD80和CD86基因敲除对成熟DC(MDCs)Th1/Th2细胞因子产生的影响却知之甚少。本研究的目的是探讨小干扰RNA(SiRNA)能否抑制MDCS表面CD80和CD86的表达。采用小鼠哮喘模型，观察CD80和CD86基因敲除对小鼠外周血中Th1/Th2细胞因子表达的影响。分离、分离和体外培养DC。流式细胞仪检测DC表面CD11c、CD80、CD86的表达。将CD80和CD86特异性siRNA导入DC，同时设计非siRNA和阴性siRNA对照。采用逆转录定量聚合酶链式反应和流式细胞术分别检测CD80和CD86的mRNA和蛋白表达水平。采用酶联免疫吸附试验检测与MDCS共培养的T细胞产生干扰素-γ和白介素4的水平。经siRNA转染后，MDCS中CD80和CD86的mRNA和蛋白水平显著下调。SiRNA组与MDCS共培养的T细胞产生的干扰素-γ水平显著升高，而IL-4的产生水平显著降低。这些结果表明，用siRNA特异性靶向CD80和CD86能够抑制CD80/CD86的表达，从而通过增加干扰素-γ的产生和降低IL-4的水平来调节Th1/Th2细胞因子水平。
Dendritic cells (DCs) are associated with the activation and differentiation of T helper (Th) cells. Cluster of differentiation (CD) 80 and CD86, the co-stimulatory molecules highly expressed in DCs, have are prominent in promoting the differentiation of Th cells toward Th2 cells. However, little is known about the effect of CD80 and CD86 knockdown on Th1/Th2 cytokine production in mature DCs (mDCs). The aim of the present study was to investigate whether small-interfering RNA (siRNA) could suppress the surface expression of CD80 and CD86 in mDCs. The effects of CD80 and CD86 knockdown in mDCs on Th1/Th2 cytokine expression were examined using an asthmatic murine model. DCs were isolated, separated and cultured in vitro. Flow cytometry was used to examine the expression of CD11c, CD80 and CD86 on the DCs. The DCs were transfected with CD80- and CD86-specific siRNA, while non-siRNA and negative siRNA controls were also designed. Then, the mRNA and protein expression levels of CD80 and CD86 were determined by reverse transcription-quantitative polymerase chain reaction and flow cytometry, respectively. The levels of interferon (IFN)-gamma and interleukin (IL)-4 produced by T cells co-cultured with mDCs were measured by enzyme-linked immunosorbent assay. Substantial downregulation of CD80 and CD86 mRNA and protein levels were observed in the mDCs following transfection with siRNA. The level of IFN-gamma produced by T cells co-cultured with mDCs was significantly increased in the siRNA group, while IL-4 production was significantly decreased. These results show that specific targeting of CD80 and CD86 with siRNA is able to suppress CD80/CD86 expression and consequently regulate Th1/Th2 cytokine levels by increasing IFN-gamma production and decreasing IL-4 levels in an asthmatic murine model.