论文部分内容阅读
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
In order to investigate the regulatory effect of interleukin-4 (IL-4) on interleukin-12 (IL-12) p70 production by dendritic cells (DCs) and its mechanism, mouse bone marrow cells 4 culture medium for 6 days, adding mature inducer lipopolysaccharide (LPS) or TNF α, CpG ODN cultured for 2 days to obtain mature DC. Flow cytometry was used to detect CD80 and MHC class II molecules on DC surface. Mixed lymphocyte reaction was used to detect the ability of DCs to promote the proliferation of allogeneic T cells. The ability of different inducers to induce IL 1 2p70 production was detected by ELISA. The expression of IL-12p35 and p4o mRNA and their changes were detected. The results showed that mature DCs can be obtained from mouse bone marrow cells cultured in culture medium containing GM CSF, IL 4 and mature inducer. CD80 and MHC class II molecules are highly expressed in mature DCs and have strong stimulation of allogeneic T cell proliferation LPS, CpG ODN, TNFα, PolyI: C can promote DC maturation and induce IL production. IL 1 2 and IL 4 can promote the production of IL-12. RT-PCR and fluorescence quantitative PCR results showed that LPS-induced IL-12 production and the role of IL-4 in its promotion were related to the increased transcription level of IL-12p35