吞噬了PUVA处理的人脾脏凋亡细胞的未成熟树突状细胞抵抗脂多糖诱导的成熟作用

发布时间：2018-06-16 20:39

本文选题：体外光化学疗法 + 树突状细胞　；参考：《细胞与分子免疫学杂志》2017年01期

【摘要】：目的探讨吞噬凋亡人脾脏淋巴细胞的未成熟树突状细胞(imDC)是否可被脂多糖(LPS)诱导成熟。方法获得人外周血单个核细胞(PBMC),利用重组人白细胞介素4(rh IL-4)、重组人粒细胞巨噬细胞集落刺激因子(rh GM-CSF)诱导生成DC。分离人脾脏淋巴细胞(h SP),经补骨脂素联合A波段紫外线(PUVA)处理h SP得到凋亡的PUVA-h SP。将imDC与PUVAh SP共培养得到体外光化学治疗性树突状细胞(ecpDC)。收集imDC、ecpDC并加入10 ng/m L的LPS培养1 d,分别得到LPS刺激的imDC和LPS刺激的ecpDC。流式细胞术检测上述细胞CD11c、CD83、CD86的表达。ELISA检测细胞上清IL-10的含量。结果 imDC和ecpDC CD83、CD86阳性率无显著性差异,LPS刺激的imDC CD83、CD86阳性率明显高于LPS处理的ecpDC。imDC培养上清液中IL-10的含量低于ecpDC,LPS刺激的imDC上清液中IL-10含量低于LPS刺激的ecpDC。结论 imDC与ecpDC均表达不成熟表型,但ecpDC可抑制LPS诱导DC成熟的作用。
[Abstract]:Objective to investigate whether immature dendritic cells (DC) phagocytosis of apoptotic human spleen lymphocytes can be induced to mature by lipopolysaccharide (LPS). Methods Human peripheral blood mononuclear cells (PBMC) were obtained and induced by recombinant human interleukin (4(rh) IL-4 and recombinant human granulocyte macrophage colony stimulating factor (rh GM-CSF). Human spleen lymphocytes were isolated and treated with psoralen combined with A band ultraviolet (PUVA) to obtain apoptotic PUVA-h SPS. In vitro photochemotherapeutic dendritic cells (DC) were obtained by co-culture of imDC and PUVAh SP. LPS-stimulated and LPS-stimulated ecpDC were obtained by collecting and adding 10 ng/m / L LPS-induced ecpDC for 1 day, and then LPS-stimulated ecpDC and LPS-stimulated ecpDC were obtained respectively. Flow cytometry was used to detect the expression of CD11cU CD83 and CD86. Elisa was used to detect the level of IL-10 in the supernatant. Results there was no significant difference between imDC and ecpDC CD83 CD86 positive rate. The positive rate of lipopolysaccharide stimulated imDC CD83 + CD86 was significantly higher than that of ecpDC.imDC supernatant treated with LPS. The level of IL-10 in imDC supernatant was lower than that in ecpDC-stimulated imDC supernatant. Conclusion both imDC and ecpDC express immature phenotypes, but ecpDC can inhibit LPS-induced DC maturation.
【作者单位】：河北北方学院研究生部;解放军第309医院全军器官移植研究所肝胆外科;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;75559部队医院;
【基金】：国家自然科学基金(811022422) 首都临床特色应用研究(Z111107058811008)
【分类号】：R392

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