CD11b激动剂leukadherin-1通过阻断NF-κB p65通路抑制小鼠骨髓来源树突状细胞TLR7和TLR9活化  

CD11b agonist leukadherin-1 inhibits activation of TLR7 and TLR9 in mouse bone marrow-derived dendritic cells by blocking NF-κB p65 pathway

作　　者：翟伟伟 邹孟君 孙明慧 梁玥 董知临 张馨元 杨永红[4] 董冠军[3] 司传平[3] ZHAI Weiwei;ZHOU Mengjun;SUN Minghui;LIANG Yue;DONG Zhilin;ZHANG Xinyuan;YANG Yonghong;DONG Guanjun;SI Chuanping(College of Basic Medicine,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117;Department of Clinical Laboratory,Jining NO.1 People's Hospital,Jining 272000;Institute of Immunology and Molecular Medicine,Jining Medical University,Jining 272067;Medical Research Center,Affiliated Hospital of Jining Medical University,Jining 272029,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)基础医学院,山东济南250117 [2]济宁市第一人民医院医学检验科,山东济宁272000 [3]济宁医学院免疫学与分子医学研究所,泰山学者实验室,山东济宁272067 [4]济宁医学院附属医院医学研究中心,山东济宁272029

出　　处：《细胞与分子免疫学杂志》2021年第1期1-7,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81771668,81601426);济宁市重点研发计划项目(2018SMNS003);济宁医学院大学生创新训练计划项目(cx2019003)。

摘　　要：目的研究整合素CD11b激动剂白细胞黏附素1(LA1)对小鼠骨髓来源的树突状细胞(BMDC)中Toll样受体7(TLR7)和TLR9通路活化的影响,并探索其调控机制。方法成功诱导BMDC,用CCK-8法和异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双染法筛选出对BMDC活性和凋亡都无影响的LA1浓度。LA1预处理BMDC 2 h后,用TLR7激动剂R837和TLR9激动剂CpG1826刺激BMDC,收集细胞用流式细胞术检测BMDC表面标志CD40、CD86和主要组织相容性复合体Ⅱ(MHC-Ⅱ)的表达;收集细胞培养上清,用ELISA检测白细胞介素6(IL-6)、IL-12p40和肿瘤坏死因子α(TNF-α)的含量;用Western blot法检测BMDC中核因子κB p65(NF-κB p65)的磷酸化水平。结果LA1浓度低于20μmol/L时对BMDC活力和凋亡无影响;LA1预处理显著抑制BMDC中R837和CpG1826诱导的CD40、CD86和MHC-Ⅱ的表达;LA1预处理显著抑制BMDC培养上清中R837和CpG1826诱导的IL-6、IL-12p40和TNF-α的含量;进一步研究发现,LA1预处理显著抑制BMDC中R837和CpG1826激活的NF-κB p65磷酸化水平。结论CD11b激动剂LA1可显著抑制BMDC中TLR7和TLR9通路的活化,且通过NF-κB p65信号通路发挥作用。Objective To study the effect of CD11b agonist leukadherin-1(LA1)on Toll-like receptor 7(TLR7)-and TLR9-induced activation of mouse bone marrow-derived dendritic cells(BMDCs)and its specific mechanism.Methods BMDCs were successfully induced and the concentrations of LA1 used in the study were determined by CCK-8 assay and annexinⅤ-FITC/PI double staining.BMDCs were treated with LA1 for 2 hours followed by stimulation of TLR7 agonist R837 and TLR9 agonist CpG1826.The expression of BMDCs surface markers CD40,CD86 and MHC-Ⅱwere detected by flow cytometry;IL-6,IL-12p40 and tumor necrosis factorα(TNF-α)in the cell culture supernatant were detected by ELISA;the phosphorylation of NF-κB p65 in BMDCs was detected by Western blotting.Results LA1 concentration below 20μmol/L had no effect on the viability and apoptosis of BMDCs.LA1 pretreatment significantly inhibited R837-and CpG 1826-induced expression of CD40,CD86 and MHC-Ⅱ,and the secretion of IL-6,IL-12p40 and TNF-αin BMDCs.Moreover,LA1 pretreatmentsignificantly inhibited the phosphorylation of NF-κB p65 activated by R837 and CpG1826 in BMDCs.Conclusion CD11b agonist LA1 can significantly inhibit the activation of TLR7 and TLR9 in BMDCs by blocking the NF-κB p65 signaling pathway.

关 键 词：骨髓来源树突状细胞(BMDC) Toll样受体(TLR) 整合素CD11b 白细胞黏附素1(LA1) 

分 类 号：R392.12[医药卫生—免疫学] R593.241[医药卫生—基础医学]