封闭PD-L1对小鼠细菌脓毒血症免疫麻痹的研究  

Effect of PD-L1 blocking on immune paralysis of mice with bacterial sepsis

作　　者：马瑜红[1] 梁婧洁 刘润秋 杨沐雨 曲震理[1] 王卫娜[1] 王方[1] 

机构地区：[1]南阳医学高等专科学校基础医学部,河南南阳473003 [2]四川农业大学农学院,四川成都611130 [3]河南中医药大学第一临床医学院,河南郑州450046

出　　处：《中华医院感染学杂志》2018年第1期10-13,共4页Chinese Journal of Nosocomiology

基　　金：河南省基础与前沿技术研究计划基金资助项目(152300410018);南阳市科技攻关计划基金资助项目(2015KJGG18)

摘　　要：目的探讨脂多糖所致细菌脓毒血症小鼠树突状细胞PD-L1的表达和免疫反应的变化。方法以脂多糖刺激小鼠树突状细胞建立脓毒血症免疫麻痹体系,利用显微拍照技术、流式细胞术、免疫荧光技术和BrdU细胞增殖反应试剂盒分别检测树突状细胞形态、PD-L1表达及抗原特异性混合淋巴细胞增殖反应的变化。结果脂多糖刺激引起树突状细胞由未成熟转变为成熟,且共抑制分子PD-L1阳性细胞平均荧光强度由Control组的4.91±0.04增加到26.56±0.39(P<0.05);抗原提呈SIINFEKL-H-2Kb阳性百分比由(19.08±0.86)%升至(87.54±1.36)%(P<0.05);抗原特异性混合淋巴细胞反应BrdU掺入实验OD值由0.767±0.026减少为0.358±0.030,但封闭PD-L1后细胞增殖反应OD值由0.358±0.030增加到0.590±0.018(P<0.05)。结论脂多糖所致脓毒血症小鼠树突状细胞高表达免疫共抑制分子PD-L1引起T细胞免疫反应低下,封闭PD-L1改善小鼠细菌脓毒血症免疫麻痹。OBJECTIVE To explore the expression of dendritic cell PD-L1 and immune response of mice with li- popolysaccharide （LPS）-induced bacterial sepsis.METHODS The dendritic cells of mice were stimulated by LPS so as to establish immune paralysis system of sepsis.The morphology of dendritic cells, expression of PI）-L1 and SI- INFEKL-specific T cell proliferation in vitro were detected by means of microscopic imaging technique, flow cy- tometry, immunofluorescence and BrdU cell proliferation assay kit.RESULTS The immature dendritic cells turned to be mature ones by the stimulation of LPS, and the average fluorescence intensity of positive PD-L1 cell was sig- nificantly increased from 4.91±0.04 in Control group to 26.56±0.39（P〈0.05） ; the percentage of positive SIIN- FEKL-H-2Kb was significantly increased from （19.08±0.86）% to （87.54±1.36）% （P〈0.05）.The OD value of BrdU-labeled SIINFEKL-specific T cell proliferation was decreased to 0.358±0.030, however, the OD value of cell proliferation was significantly increased from 0.358 ± 0. 030 to 0.590 ± 0. 018 after the PD-L1 blocking （P 0.05）.CONCLUSION The high-expression co-inhibitory molecule PD-L1 in the dendritic cells may lead to the low immune response of T cells of the mice with LPS-induced sepsis, however, the PD-L1 blocking may improve the immune paralysis of the mice with bacterial sepsis.

关 键 词：PD-L1 脂多糖 细菌脓毒血症 感染 免疫麻痹 

分 类 号：R631.1[医药卫生—外科学]