MyD88在OK-432诱导树突状细胞成熟中的作用  

Effect of MyD88 on Maturation of OK-432-Stimulated Dendritic Cells

作　　者：倪秀雄[1] 刘丽燕[1] 刘永建 曾真[1] 袁明洲[3] 陈玄[4] 

机构地区：[1]福建医科大学基础医学院生理学与病理生理学系,福州350004 [2]河北省赵县人民医院,赵县050130 [3]福建医科大学附属第一医院血液科,福州350005 [4]福建省中医药研究院,福州350001

出　　处：《福建医科大学学报》2010年第1期28-31,共4页Journal of Fujian Medical University

基　　金：福建省自然科学基金(C0410020)

摘　　要：目的探讨小鼠骨髓树突状细胞(DCs)髓性分化蛋白88(MyD88)在OK-432诱导DCs成熟中的作用机制。方法分离小鼠骨髓单核细胞(BMMCs),在含10%胎牛血清、重组小鼠粒-巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白细胞介素-4(rmIL-4)的培养体系中诱导培养,所得的细胞即为DCs。于培养的第5天,将细胞分为3组:对照组不加OK-432和MyD88 si RNA;OK-432组加入终浓度为5μg/mL的OK-432;RNA干扰组加入MyD88 si RNA12 h后,再加入5μg/mL OK-432刺激。半定量RT-PCR法检测DCs中MyD88的表达;ELISA法检测各组DCs分泌肿瘤坏死因子-α(TNF-α)和IL-12的能力;MTT法检测DCs体外刺激同种混合淋巴细胞增殖的免疫活性。结果OK-432组DCs的MyD88高表达、TNF-α和IL-12释放明显增加并能诱导较强混合淋巴细胞反应,而用MyD88 si RNA干扰后以上效应均降低(P<0.05)。结论靶向MyD88 si RNA可明显抑制小鼠DCs中MyD88的表达和OK-432诱导的DCs成熟,表明MyD88介导OK-432诱导的DCs成熟。Objective To investigate the effect of myeloid differentiation protein 88 （MyD88） siRNA on OK-432-stimulated dendritic cells （DCs） from mouse bone marrow. Methods Mouse bone marrow mononuctear celts （BMMCs） were cultured in RPMI1640 containing 10% FCS, rmGM-CSF, and rmIL-4. The cultured cells were divided into three groups on the fifth day. To the control group, noth- ing was added; to the OK-432 group, OK-432 at a final concentration of 5μg/mL was added; to the RNA intervention group, MyD88 siRNA was added and 12 hours later OK-432 at a final concentration of 5 μg/mL was added. All groups were further cultured for 48 hours. The expression of MyD88 was measured by RT-PCR. The concentration of TNF-α and IL-12 in the supernatant was detected by ELISA. Antigen presenting ability of DCs in allo-MLR was measured by MTT assay. Results In the OK-432 group, there was a high expression of MyD88, increased concentration of TNF-α and IL-12 in the supernatant were increased, and enhanced allo-stimulating capacity. MyD88 siRNA inhibited all these responses. Conclusion MyD88-targeted siRNA can reduce MyD88 expression in mouse myeloid DCs and inhibit the maturation of OK-432-stimulated DCs, which indicates that OK-432 may induce the maturation of DCs through TLRs/MyD88 signal pathway.

关 键 词：树突状细胞 RNA 小分子干扰 毕西巴尼 免疫耐受 

分 类 号：R392[医药卫生—免疫学]