IL-4对DC产生IL-12影响的研究

为了研究白细胞介素 4 (IL 4 )对树突状细胞 (dendriticcell,DC )产生白细胞介素 1 2 (IL 1 2 )p70的调节作用及其机制 ,将小鼠骨髓细胞在含GM CSF和IL 4的培养液中培养 6d ,加入成熟诱导剂脂多糖 (LPS )或TNF α、CpG ODN继续培养 2d以获得成熟DC。流式细胞仪检测DC表面CD80和MHCII类分子 ,混合淋巴细胞反应检测DC促进同种异体T细胞增殖的能力 ,ELISA法检测不同诱导剂诱导DC产生IL 1 2p70的能力 ,RT PCR和荧光定量PCR检测IL 1 2p35和p4 0mRNA的表达及其变化。结果显示小鼠骨髓细胞在含GM CSF、IL 4和成熟诱导剂的培养液中培养后可以获得成熟的DC ,成熟DC高表达CD80和MHCII类分子 ,具有较强的刺激同种异体T细胞增殖的能力 ,LPS、CpG ODN、TNF α、PolyI:C在促进DC成熟的同时能诱导DC产生有活性的IL 1 2 ,IL 4对IL 1 2的产生具有明显的促进作用 ,RT PCR和荧光定量PCR结果显示LPS诱导IL 1 2产生以及IL 4对其的促进作用与IL 1 2p35基因的转录水平增高有关

Effects of IL-4 on the Production of IL-12 by Dendritic Cells

To evaluate the effect of IL 4 on the ability of dendritic cells(DC) to produce IL 12 p70 and its regulatory mechanism, the mature mouse bone marrow derived DC were generated by cultivation of C57BL/6 mice bone marrow cells with GM CSF and IL 4 for 6 days and followed by addition of LPS ,TNF α or CpG ODN for 2 days The surface molecules of DC including CD80 and MHC class II molecules were analyzed by FACS, the stimulatory effect of DC on the proliferation of allogenic T lymphocytes was examined by 3 H TdR incorporation assay, and IL 12 p70 was quantified by enzyme immunoassay IL 12 p35 and p40 mRNA expressions and its changes were measured by RT PCR and real time PCR It was found that the mature DC could be obtained by these methods of study These mature DC showed a high level of expressions in CD80 and MHC class II molecules, and have capacity to stimulate proliferation of the naive T cells in vitro IL 4 strongly enhanced the production of bioactive IL 12 p70 in the presence of maturation stimulus Results by using RT PCR and real time PCR indicated that the effects of IL 4 on the production of IL 12 p70 might be mediated by the regulation of IL 12 p35 mRNA