吞噬PUVA处理的人脾细胞的未成熟树突状细胞具有调节性树突状细胞特性  

Immature dendritic cells phagocytosing human spleen cells treated by PUVA present the characteristics of regulatory dendritic cells

作　　者：杨绍臻 杜国盛[1] 魏玉香[2] 索龙龙[1] 周林[1] 封立魁[1] 宋继勇[1] 朱志东[1] 郑永根 

机构地区：[1]解放军第309医院全军器官移植研究所肝胆外科 [2]解放军第309医院全军器官移植研究所器官移植与免疫调节北京市重点实验室 [3]75559部队医院

出　　处：《细胞与分子免疫学杂志》2017年第4期455-459,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(811022422);首都临床特色应用研究(Z111107058811008)

摘　　要：目的探讨吞噬经补骨脂素长波紫外线(PUVA)处理的人脾淋巴细胞的未成熟树突细胞(imDC)的表型及功能。方法分离人外周血单个核细胞(PBMC),利用白细胞介素4(IL-4)和重组人粒细胞、巨噬细胞集落刺激因子诱导培养。培养第6天收获imDC;其中一部分加入脂多糖(LPS)1 d诱导为成熟DC。PUVA处理人脾脏淋巴细胞(PUVA-SP)分别将PUVA-SP、SP与imDC共培养,获得体外光化学治疗性树突状细胞(ecpDC)和SP-DC。收集各组DC,检测其表面抗体CD11c、CD83、CD86的表达情况。获取上述细胞上清,ELISA检测IL-10和IL-12含量。结果经PUVA处理后的SP早期凋亡率为(94.21±3.75)%。SP-DC组CD83、CD86的阳性率明显高于imDC,而ecpDC组CD83、CD86的阳性率与imDC相近,但低于DC。结论吞噬凋亡的SP得到的DC为imDC,具有负向免疫调控作用。Objective To investigate the effect of psoralen combined with A-band ultraviolet （PUVA）-treated human spleen lymphocytes on the phenotype and function of immature dendritic cells （imDCs）. Methods Human peripheral blood mononuclear cells （PBMCs） were isolated and induced to produce DCs by interleukin-4 （IL-4） and recombinant human granulocyte macrophage colony stimulating factor （rhGM-CSF）. On the sixth day, the imDCs were obtained and stimulated by lipopolysaccharide （LPS）. One day later, mature DCs were harvested. Human spleen cells （SPs） were isolated and treated with PUVA to prepare apoptotic PUVA-SPs. Co-culture of imDCs with PUVA-SPs resulted in extracorporeal photochemotheraputic DCs （ecpDCs）. Co-culture of imDCs with SPs resulted in SP-DCs. The expressions of CD11c, CD83 and CD86 were detected by flow cytometry. The levels of IL-10 and IL-12 in the supernatants of the above cells were determined by ELISA. Results The early apoptosis rate of PUVA-SPs was （94.21 ± 3. 75 ） %. There was no significant difference in the expressions of CD83 and CD86 between imDCs and ecpDCs. But the positive rates of CD83 and CD86 in ecpDCs were lower than those in DCs. However, the positive rates of CD83 and CD86 in SP-DCs were significantly higher than those of the imDCs. Conclusion The imDCs phagocytosing apoptotic human SPs present phenotype and function of regulatory DCs.

关 键 词：体外光化学疗法 树突状细胞 细胞表型 免疫耐受 

分 类 号：R392.1[医药卫生—免疫学] R454.2[医药卫生—基础医学]