中文摘要：

目的观察细粒棘球蚴重组抗原B（rAgB）体外诱导小鼠骨髓源树突状细胞表达吲哚胺2,3-双加氧酶（IDO）的情况。方法从小鼠股骨中分离出骨髓细胞,进行小鼠重组巨噬细胞集落刺激因子（rmGM-CSF）的诱导,培养并获得CD11c＋树突状细胞。应用倒置显微镜和扫描电镜观察树突状细胞形态,采用流式细胞术检测其表面标志物;用混合淋巴细胞反应（MLR）观察树突状细胞对T淋巴细胞的增殖能力。培养至第6天,收集未成熟树突状细胞进行流式细胞术检测;另向部分未成熟树突状细胞中加脂多糖（LPS）刺激24 h后,收集成熟树突状细胞,进行流式细胞术检测。在获得的未成熟树突状细胞中分别加入RPMI 1640完全培养液（为阴性对照组）、重组小鼠γ干扰素（rmIFN-γ,1 000 U/ml,为IFN-γ组）和rAgB（终浓度15μg/ml,为rAgB组）,培养24 h后,通过细胞免疫组织化学和蛋白质印迹（Western blot-ting）检测各组树突状细胞IDO的表达情况。结果获得纯度为80%的CD11c＋树突状细胞,在倒置显微镜和扫描电镜下均观察到典型树突状细胞。经LPS刺激的成熟树突状细胞的CD40、CD80和主要组织相容性复合体（MHC）Ⅱ类分子I-A/I-E的阳性表达率与未成熟树突状细胞的相比,差异均有统计学意义（均P〈0.05）。MLR结果显示,诱导形成的树突状细胞具有刺激T淋巴细胞增殖的能力。细胞免疫组织化学方法检测结果显示,阴性对照组、阳性对照组和rAgB组的IDO阳性表达率分别为（4.544±1.752）%、（20.464±4.452）%和（11.148±1.966）%,3组间的差异均有统计学意义（均P〈0.05）。Western blotting结果表明,3组IDO蛋白与相应甘油醛-3-磷酸脱氢酶（GAPDH）蛋白的灰度值之比分别为（0.229±0.085）、（0.794±0.114）和（0.573±0.129）,其中rAgB组与阴性对照组相比差异有统计学意义（P〈0.05）,与IFN-γ组相比差异无统计学意义（P〉0.05）。结论细?

英文摘要：

Objective To observe the expression of indoleamine 2,3-dioxygenase（IDO） in mouse bone marrow-derived dendritic cells（DCs） after adding Echinococcus granulosus recombinant antigen B（rAgB） in vitro.Methods CD11c＋ DCs generated from bone marrow precursor cells of C57BL/6 mice and cultured in the presence of recombinant mouse GM-CSF（rmGM-CSF）.The morphology of DCs was observed by inverted microscope and scanning electronic microscope.The level of I-A/I-E,CD40,CD80,and CD86 on DCs were determined by flow cytometry.T cell proli-feration induced by DCs were evaluated by using mixed lymphocyte reaction（MLR） assay.At day 6 postculture,the imm-ature DCs were collected,and part of the immature DCs stimulated with lipopolysaccharide（LPS） for 24 h were exa-mined by flow cytometry.Immature DCs were divided into 3 groups： negative control group,positive control group（rmIFN-γ,1 000 U/ml） and rAgB group.Immature DCs of positive control group and rAgB group were induced with 1 000 U/ml rmIFN-γ and 15 μg/ml rAgB,respectively.IDO expression in DCs was examined 24 h after induction using immunohistochemical method and Western blotting.Results More than 80% CD11c＋ DCs were harvested.The typical DCs were observed under inverted microscope and scanning electronic microscope.The level of CD40,CD80,and IA/IE（MHCⅡ） in mature DCs group was significantly higher than that of immature DCs group（P0.05）.In MLR,mitomycin-treated DCs can stimulate T lymphocytes proliferation activity.There were significantly differences in IDO expression in the negative control group [（4.544±1.752）%],positive control group [（20.464±4.452）%] and rAgB group [（11.148±1.966）%]（P0.05）.Western blotting result indicated that the ratio of IDO/GAPDH in rAgB group（0.573±0.129） was significantly higher than that of negative group（0.229±0.085）（P0.05）,and there were no significant difference in the ratio of IDO/GAPDH between IFN-γ group（0.794±0.114） and rAgB group（P0.05）.Conclusion rAgB c