›› 2013, Vol. 31 ›› Issue (3): 6-188-192.

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Different Echinococcus granulosus Antigens Induced Indoleamine 2，3-dioxygenase Expression in Dendritic Cells

SHAN Jiao-yu1,2, LI Hai-tao1,3, LI Chun-yan2, XIAO Jin2, LI Liang1, ZHANG Xue1, LIN Ren-yong1, WEN Hao1,3 *   

  1. 1 State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China; 2 Department of Human Parasitology, School of Basic Medicine, Xinjiang Medical University, Urumqi 830054, China; 3 Department of Hepatobiliary Diseases, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China
  • Online:2013-06-30 Published:2013-07-17

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Abstract: 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-A/I-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 IDO 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±3.54）%, （53.00±3.39）%,　（77.10±1.70）%］ 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）］（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 Try were lowest in rAgB-treated group ［（23.65±0.64）, （13.95±1.06）, （19.05±0.64） μmol/L］. At 24 h after induction, Try concentration in negative control group（22.90±0.14） was higher than that of MHF-treated group （20.65±0.34）（P<0.05）.  Conclusion  Under in vitro condition, rAgB and MHF can up-regulate IDO expression. The ability of rAgB to up-regulate IDO activity was stronger than that of MHF at 24 h after induction.