中文摘要：

目的用核因子κB（NF-κB）寡核苷酸诱骗剂（ODN Decoy）诱导大鼠耐受型树突状细胞（DC）并负载牛Ⅱ型胶原（BⅡC）,评价其免疫学特性。方法用GM-CSF和IL-4诱导SD大鼠脾脏单个核细胞分化成DC,将其分为对照组及实验1、2组,均继续加入GM-CSF、IL-4,同时实验1、2组加入ODN Decoy,培养7 d实验2组加入BⅡC。流式细胞术检测细胞表面共刺激分子CD80、CD86,同种异体淋巴细胞刺激试验评价淋巴细胞增殖情况及其对BⅡC和人血清刺激的反应,检测上清液中Th1/Th2细胞因子IFN-γ/IL-10。结果与对照组相比,实验1、2组CD80、CD86表达均降低（P均〈0.05）;实验1组淋巴细胞增殖程度及IFN-γ明显降低（P均〈0.05）,IL-10无明显变化;实验2组加入BⅡC后淋巴细胞增殖程度及IFN-γ、IL-10均无明显变化（P均〉0.05）。实验2组中,与加入BⅡC相比,加入健康人血清后淋巴细胞明显增殖（P〈0.05）,IFN-γ升高,IL-10降低（P均〈0.05）。结论用NF-κB ODN Decoy成功诱导获得耐受型DC。该DC刺激同种异体淋巴细胞反应的能力较低,且抑制淋巴细胞免疫反应向Th1方向偏移;负载BⅡC后刺激的淋巴细胞对该抗原特异性耐受,而对无关抗原的免疫应答无明显影响。

英文摘要：

Objective To evaluate the immunological characters of rat tolerogenic dendritic cells（ DCs） induced by nuclear factor κB oligonucleotides decoy（ NF-κB ODN decoy） and loaded with bovine collageb typeⅡ（ BⅡC） in vitro. Methods The rat spleen monocytes were induced by recombinant GM-CSF and IL-4 in vitro to differentiate into DCs. Those DCs were divided into three groups： the control group which was added with cytokines,experimental group 1 which was dealt with NF-κB ODN decoy,and the experimental group 2 which was dealt with NF-κB ODN decoy and followed by BⅡC after being cultured for7 days. The expression of CD80 and CD86 was analyzed by using flow cytometry. Allogeneic lymphocyte stimulation test was used to evaluate the lymphocyte proliferation and the reaction for BⅡC and human serum stimulation. Meanwhile,the concentration of IL-10 and IFN-γ in supernatant was detected. Results Compared with the control group,the CD80 and CD86 expression intensities in the experimental groups 1 and 2 were decreased（ all P〈0. 05）; the lymphocyte proliferation and expression of IFN-γ was significantly decreased in the experimental group 1（ all P〈0. 05）,but the IL-10 showed no significant difference. In the experimental group 2,after adding BⅡC,lymphocyte proliferation and expression of IFN-γ and IL-10 showed no significant difference（ all P〈0.05）. However,after adding healthy human serum,the lymphocyte proliferation and expression of IFN-γ was significantly increased,and the expression of IL-10 was decreased in the experimental group 2（ all P〈0. 05）. Conclusions NF-κB ODN decoy successfully induced to construct rats＇ tolerogenic DCs. The tolerogenic DC exhibited low capability to stimulate lymphocytes and inhibited the lymphocyte immune response to Th1 shift direction. When loaded with BⅡC,the lymphocytes that stimulated by BⅡC exhibited specific tolerance against the specific antigens but had no significant effect on the antigen-independent immune response.