补体调节蛋白Crry对树突状细胞的调控及诱导同种移植免疫低反应  

Complement regulatory protein Crry regulates dendritic cells and induces allograft immune hyporesponsiveness

作　　者：张艳[1] 陈栋[1,2] 李明[1,2] 李永海[1,2] 刘斌[1,2] 陈刚[1,2] 陈实[1,2] 

机构地区：[1]华中科技大学同济医学院附属同济医院综合医疗科,武汉430030 [2]华中科技大学同济医学院附属同济医院器官移植研究所,卫生部/教育部器官移植重点实验室,武汉430030

出　　处：《中国免疫学杂志》2013年第1期25-28,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.30600573)

摘　　要：目的:探讨补体调节蛋白Crry对树突状细胞(DC)的调节作用及诱导同种移植免疫低反应性的机制。方法:分离BALB/c小鼠的骨髓来源树突状细胞,试验分为2组:①脂多糖(LPS)刺激组,即培养结束前加入外源性脂多糖(1 mg/L)刺激24小时;②Crry处理组,加入抗小鼠Crry抗体5μg/ml,结束前加入外源性脂多糖(1 mg/L)刺激24小时,流式细胞仪检测树突状细胞表面分子CD86、CD40、MHCⅡ的表达,ELISA法检测其细胞上清液中IFN-γ、IL-10和IL-12的水平,ELISA法检测细胞上清液中C3、C5、C3a和C5a的含量,并以BALB/c小鼠的DC作为刺激细胞,以C57BL/6小鼠的淋巴细胞作为反应细胞进行同种混合淋巴细胞培养,MTT法检测淋巴细胞增殖活性。结果:经LPS刺激后,培养的DC表面共刺激分子CD40、CD86和MHCⅡ分子的表达显著增加(P<0.05);Crry处理组DC表面共刺激分子CD86和MHCⅡ分子的表达较LPS刺激组显著降低(P<0.05),而CD40的表达无显著性差异;Crry处理组培养上清液中C3和C5的含量无明显变化,但C3a、C5a的含量较LPS刺激组显著减少(P<0.05);培养DC上清液中IFN-γ和IL-12的含量较LPS刺激组显著降低(P<0.05),IL-10的含量较LPS刺激组显著增加(P<0.05);培养的DC加入Crry抗体后,其淋巴细胞增殖活性显著降低(P<0.05)。结论:补体调节蛋白Crry可以对DC具有重要的调控作用,可以影响其共刺激分子的表达、补体的生成以及细胞因子的合成等,从而诱导同种移植免疫低反应性,丰富先天免疫对获得性免疫的调节作用。Objective ：To investigate the allograft immune hyporesponsiveness induced by complement regulatory protein Crry as well as dendritic cells （DCs） involved. Methods： DCs were isolated and cultured from bone marrow of BALB/c mice. The experi- ment was divided into 2 groups, （1）LPS-stimulated group, DCs were cultured and stimulated by LPS for 48 h ; （2） Crry-treated group, DCs were cultured and anti-Crry antibody were added to culture medium with the concentration of 5 μg/ml, DCs were stimulated by LPS for 48 h. The expression of CD86, CIM0, MHC lI in DCs were examined by flow cytometry. Production of interferon-γ（IFN-γ） , interleukin-10（IL-10） and interleukind2（IL-12） was detected in the supernants of different groups by ELSIA; complement C3, C5, C3a and C5a in the supernants of different groups were detected by ELISA; Allo-MLR was examined by MTT colorimetry with DCs from BALB/c mice as the stimulators and CD4 ＋T cells of C57BL/6 mice as responders. Results：Compared with LPS-stimulated group, CD86 and MHC II expression in DCs , C3a, C5a,IFN-γ/and IL-12 in the upernants were significantly decreased（P 〈 0.05）, IL-10 in the supernants was significantly increased （ P 〈 0.05 ）, Crry-stimulated DCs significantly inhibited proliferation in mixed lymphocyte culture （ P 〈 0.05）. Conclusion ： Complement regulatory protein Crry modulated costimulatory molecular expression, cytokines produc- tion, and complement production in DCs, which induces allograft immune hyporesponsiveness.

关 键 词：补体调节蛋白 Crry 树突状细胞 同种移植 

分 类 号：R392.4[医药卫生—免疫学]