OCILRP2-Fc抑制小鼠骨髓来源的树突状细胞分化成熟

发布时间：2018-10-31 21:27

【摘要】：目的:采用OCILRP2胞外段蛋白OCILRP2-Fc阻断OCILRP2受体,探讨OCILRP2对小鼠树突状细胞发育成熟的影响及其机制。方法:分离培养小鼠骨髓来源的树突状细胞(BMDC),荧光定量RT-PCR和流式细胞术分析OCILRP2在不成熟BMDC和LPS诱导的成熟BMDC的表达差异;流式细胞术分析OCILRP2-Fc对BMDC细胞表面标志分子的表达以及对BMDC抗原摄取能力的影响;ELISA检测培养上清中细胞因子的浓度,分析OCILRP2-Fc对BMDC分泌炎性细胞因子的影响;Western blot检测转录因子NF-κB的活化,分析OCILRP2-Fc影响BMDC分化成熟的分子机制。结果:荧光定量RT-PCR和流式细胞术结果均表明LPS可以显著上调OCILRP2在BMDC的表达(P0.05);与对照组相比,OCILRP2-Fc可以明显抑制MHCⅡ类分子及共刺激分子CD80在LPS诱导的成熟BMDC的表达;和成熟BMDC相比,OCILRP2-Fc组BMDC经LPS诱导24 h后仍然具有较强的抗原吞噬能力;而且,OCILRP2-Fc组BMDC培养上清中炎性细胞因子IL-6、IL-12、TNF-α的浓度明显低于对照组(P0.05)。Western blot结果表明OCILRP2-Fc抑制了LPS诱导的I-κB降解和NF-κB p65的磷酸化。结论:OCILRP2-Fc阻断OCILRP2受体信号,通过抑制LPS诱导的NF-κB活化影响小鼠BMDC分化成熟。提示OCILRP2受体在树突状细胞的分化成熟过程中具有促进作用。
[Abstract]:Aim: to investigate the effect of OCILRP2 on the development and maturation of mouse dendritic cells by blocking OCILRP2 receptor with OCILRP2 extracellular segment protein OCILRP2-Fc. Methods: (BMDC), quantitative RT-PCR and flow cytometry were used to analyze the difference of OCILRP2 expression in immature BMDC and LPS induced mature BMDC. Flow cytometry was used to analyze the effect of OCILRP2-Fc on the expression of marker molecules on the surface of BMDC cells and on the ability of BMDC antigen uptake, ELISA was used to detect the concentration of cytokines in culture supernatant, and the effect of OCILRP2-Fc on the secretion of inflammatory cytokines by BMDC was analyzed. The activation of transcription factor NF- 魏 B was detected by Western blot, and the molecular mechanism of OCILRP2-Fc affecting the differentiation and maturation of BMDC was analyzed. Results: fluorescence quantitative RT-PCR and flow cytometry showed that LPS could significantly up-regulate the expression of OCILRP2 in BMDC (P0.05). Compared with the control group, OCILRP2-Fc could significantly inhibit the expression of MHC class 鈪，

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