Immune modulation by silencing IL-12 production in dendritic cells using small interfering RNA
Immune modulation by silencing IL-12 production in dendritic cells using small interfering RNA
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DOI：
10.4049/jimmunol.171.2.691
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发表时间：
2003-07-15
影响因子：
4.4
通讯作者：
Min, WP
中科院分区：
文献类型：
--
作者：
Hill, JA;Ichim, TE;Min, WP
RNA干扰是一种转录后基因沉默机制，在大多数真核细胞中发挥作用，包括人类和小鼠。特异性基因沉默是由长度约为21 nt的短链双链RNA（称为小干扰RNA或siRNA）介导的，其靶向同源mRNA序列进行降解。我们在这里证明，RNAi可以用于免疫调节靶向树突状细胞（DC）基因表达。用IL-12 p35基因特异性siRNA转染DC导致基因表达的有效抑制和生物活性IL-12 p70产生的阻断，而不影响无关基因或细胞活力。IL-12的抑制与IL-10产生的增加相关，这赋予DC在体外刺激同种异体T细胞产生Th 2细胞因子的能力。此外，缺乏IL-12产生的SiRNA沉默DC在MLR中是较差的同种刺激因子。IL-12沉默和KLH脉冲DC极化的免疫反应向Th 2细胞因子的Ag特异性方式。这些数据首次证明RNA干扰是调节DC介导的免疫应答的有效和特异性工具。
RNA interference is a mechanism of posttranscriptional gene silencing that functions in most eukaryotic cells, including human and mouse. Specific gene silencing is mediated by short strands of duplex RNA of similar to21 nt in length (termed small interfering RNA or siRNA) that target the cognate mRNA sequence for degradation. We demonstrate here that RNAi can be used for immune modulation by targeting dendritic cell (DC) gene expression. Transfection of DC with siRNA specific for the IL-12 p35 gene resulted in potent suppression of gene expression and blockade of bioactive IL-12 p70 production without affecting unrelated genes or cellular viability. Inhibition of IL-12 was associated with increased IL-10 production, which endowed the DC with the ability to stimulate production of Th2 cytokines from allogenic T cells in vitro. Furthermore, siRNA-silenced DC lacking IL-12 production were poor allostimulators in MLR. IL-12-silenced and KLH-pulsed DC polarized the immune response toward a Th2 cytokine profile in an Ag-specific manner. These data are the first to demonstrate that RNA interference is a potent and specific tool for modulating DC-mediated immune responses.