中文摘要：

目的检测不同的棘球蚴抗原体外诱导树突状细胞（DCs）表达吲哚胺2，3．双加氧酶（IDO）的情况。方法从C57BL／6小鼠股骨中分离出骨髓细胞，用小鼠重组巨噬细胞集落刺激因子（rmGM．CSF）诱导后，获得小鼠骨髓源树突状细胞（BMDCs）。分别用重组抗原B（rAgB，15μg／ml）、小鼠细粒棘球蚴囊液（MHF，5mg／ml）、γ干扰素（IFN-γ，1000U／ml，为阳性对照）和RPMI1640完全培养液（阴性对照）刺激DCs，于18、24和48h后收集细胞和细胞上清。采用流式细胞术检测各组DCs的表面标志物CD40、CD80、CD86和I-MI—E的阳性表达情况，并利用实时荧光定量PCR（FQ。RT—PCR）检测各组的IDOmRNA相对转录水平。利用高效液相色谱法（HPLC）检测各组细胞上清中的色氨酸（Try）浓度。结果流式细胞术检测结果显示，DCs受rAgB和MHF刺激后，其表面标志物CD40、CD80、CD86和I-A／I—E的阳性表达率均降低。刺激24h后，rA邸组的CD40、CD86和I-A／I-E阳性表达率分别为（22．60±2．69）％、（35．50±4．38）％和（57．30±4．38）％，与MHF组[（38．00±3．54）％、（53．00±3．39）％和（77．10±1．70）％]和阴性对照[（37．95±3．61）％、（19．55±1．06）％和（85．45±1．63）％]的差异均有统计学意义（P〈0．05）。FQ—RT—PCR结果显示，刺激18、24和48h后，rAgB组的IDOmRNA水平『（9．20±0．01）、（29．44±0．02）和（16．48±0．04）]和MHF组的[（9．67＋0．02）、（17．52＋0．01）和（16．81＋0．01）]均高于阴性对照组[（2．46±0．01）、（7．77±0．01）和（10．56＋0．01）]（P〈O．01），rAgB组和MHF组IDOmRNA水平的差异均有统计学意义（P〈0．05）。HPLC结果显示，刺激18、24和48h后，rAgB组DCs上清中的色氨酸浓度[（23．65±0．64）、（13．95±1．06）和（19．05±0．64）μmol／L]均较其他3组低．刺激24h后的色氨酸浓度与阴性对照?

英文摘要：

Objective To observe the expression of indoleamine 2, 3-dioxygenase （IDO） in dendritic cells （DCs） via different Echinococcus granulosus antigens in vitro. Methods Bone Marrow DCs generated from bone marrow precursor cells of C57BL/6 mice and cultured in the presence of recombinant mouse GM-CSF （rmGM-CSF）. Then, DCs were induced with 15 μg/ml recombinant antigen B （rAgB）, 5 mg/ml mouse hydatid fluid （MHF）, 1 000 U/ml IFN-γ （as positive control）, and RPMI 1640 complete medium （as negative control）, respectively. Meanwhile, the treated DCs and cell supernatants were collected at 18, 24 and 48 h after induction. The positive expressions of D40, CD80, CD86 and I-MI-E on DCs were determined by flow cytometry. By real-time fluorescent quantitative reverse-transcription polymerase chain reaction （FQ-RT-PCR）, the expression level of 1DO mRNA in DCs was measured. Concentrations of tryptophan （Try） were tested by high-performance liquid chromatography （HPLC） assay in cell supernatant. Results The data from flow cytometry showed that the positive expressions of CD40, CD80, CD86, I-A/I-E were decreased after stimulated by rAgB and MHF. At 24 h after induction, there was significant difference in the level of CD40, CD86 and I-A/I-E among rAgB-treated group [（22.60±2.69）%, （35.50±4.38）%, （57.30±4.38）%], MHF-treated group [（38.00±.54）%, （53.00± 3.39）%, （77.10±1.70）%1 and negative control [（37.95±3.61） %, （19.55±1.06） % and （85.45±1.63） %（P〈0.05）. At 18, 24 and 48 h after induction, the levels of IDO mRNA in rAgB-treated group [（9.20±0.01）, （29.44±0.02）, （16.48± 0.04）] and MHF-treated group [（9.67±0.02）, （17.52±0.01）, （16.81±0.01）] was higher than that of negative control group[（2.46±0.01）, （7.77±0.01）, and（10.56-＋0.01）l（P〈0.01）. And significant difference was found between rAgB-treated group and MHF-treated group （P〈0.05）. At 18, 24 and 48 h after induction, the concentrations of