树突状细胞介导rEg.mMDH产生的免疫应答研究  

Effect of the recombinant antigen mMDH on the immune response induced in dendritic cells

作　　者：王双 王娅娜[1,2] WANG Shuang;WANG Ya-na(School of Basic Medicine,Ningxia Medical University,Ningxia,Yinchuan 750001,China;Key Laboratory of Hydatidosis in Ningxia Medical University)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750001 [2]宁夏医科大学包虫病重点研究室

出　　处：《中国病原生物学杂志》2021年第2期187-192,共6页Journal of Pathogen Biology

基　　金：宁夏自然科学基金项目(No.2020AAC03160);2018年大学生创新创业训练计划项目(自治区级)(No.NXCX2018149)。

摘　　要：目的观察重组细粒棘球绦虫抗原mMDH(rEg.mMDH)诱导树突状细胞(Dendritic cells,DC)产生的免疫应答。方法用rEg.mMDH体外致敏小鼠骨髓DC细胞,分为空白对照组、HF组、rEg.mMDH组、LPS组、HF+LPS组和rEg.mMDH+LPS组。电镜观察各组DC的形态;流式细胞仪检测各组DC的吞噬功能及表面分子CD80、CD86、主要组织相容性复合体II(MHCⅡ)及CD40表达情况;酶联免疫吸附试验(ELISA)检测各组DC培养上清液中细胞因子白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-12p70(IL-12p70)及TNF-α的含量。结果优化条件筛选出rEg.mMDH作用DC的最佳浓度为1μg/ml,最佳时间为20 h。电镜观察rEg.mMDH和HF组DC具有突起的细胞数目少;DC表面分子CD80、MHCII表达水平与空白对照组比较差异无统计学意义(P>0.05);且具有良好的吞噬能力。当加入LPS作用后,rEg.mMDH组DC表面的突起增多,含突起的DC数目从30%增加至52%,吞噬能力降低,MHCII、CD80、CD40表达量不同程度增高;ELISA检测rEg.mMDH组TNF-α、IL-6、IL-10含量高于空白对照组及LPS组(均P<0.01),与LPS联合作用后IL-6、IL-10显著降低(P<0.05),IL-12p70含量显著升高(P<0.01);HF组与HF+LPS组各细胞因子含量变化差异无统计学意义(P>0.05)。结论rEg.mMDH促进DC的成熟能力较弱,但刺激后的DC具有良好的吞噬功能,通过释放高含量的IL-10、IL-6,可能激活Th2型免疫应答;联合LPS,诱导DC成熟的能力增强,高表达IL-12p70并降低IL-10、IL-6的含量。Objective To explore the immune response induced in dendritic cells(DCs)by the recombinant antigen rEg.mMDH.Methods The bone marrow DCs of mice were stimulated with rEg.mMDH in vitro and divided into a control group,an HF group,an rEg.mMDH group,an LPS group,an HF+LPS group,and an rEg.mMDH+LPS group.The morphology of DCs in each group was observed using an electron microscope.Expression of the phagocytosis and surface molecules CD80,CD86,MHC-II,and CD40 by DCs was examined using flow cytometry.The levels of IL-6,IL-10,IL-12 p70,and TNF-a in the supernatant of DCs in each group were determined using ELISA.Results The optimal concentration of rEg.mMDH affecting DCs was 1μg/mL and the optimal time was 20 h.Electron microscopy indicated that DCs had fewer and shorter protrusions on their surface in the rEg.mMDH and HF groups than in other groups except for the control group.There were no significant differences in the levels of expression of CD80 and MHCII on the surface of DCs compared to levels in the control group(P>0.05),and the DCs in those groups displayed good phagocytic ability.In the rEg.mMDH group,the number of DCs with projections increased from 30% to 52% after the addition of LPS,but their phagocytic ability decreased,and the level of MHCII,CD80,and CD40 expression increased to different degrees.ELISA indicated that the levels of TNF-a,IL-6,and IL-10 in the rEg.mMDH group were higher than those in control group and the LPS group(P<0.01).The levels of IL-6 and IL-10 in the group receiving LPS as well decreased significantly,and the levels of IL-12 p70 increased significantly(P<0.01).There were no significant differences in cytokine levels in the HF group and HF+LPS group(P>0.05).Conclusion rEg.mMDH has a weak ability to promote DC maturation and it has good phagocytic action.It may activate a Th2 type immune response by releasing high levels of IL-10 and IL-6.When combined with LPS,rEg.mMDH induced the maturation of DCs,IL-12 p70 was highly expressed,and levels of IL-10 and IL-6 decreased.

关 键 词：rEg.mMDH 树突状细胞 免疫应答 

分 类 号：R383.33[医药卫生—医学寄生虫学]