Dectin-2受体介导宿主抵抗烟曲霉菌感染的研究  

Dectin-2 mediates host defense against Aspergillus fumigatus infection

作　　者：吴梦雪 于垚 郭文 王蓉蓉 贾鑫明[1] WU Mengxue;YU Yao;GUO Wen;WANG Rongrong;JIA Xinming(School of Medicine,Tongji University,Shanghai 200092,China)

机构地区：[1]同济大学医学院,上海200092

出　　处：《同济大学学报（医学版）》2022年第5期626-631,共6页Journal of Tongji University(Medical Science)

基　　金：上海市科学技术委员会基础研究重点基金(20JC1417700)。

摘　　要：目的探讨C型凝集素受体Dectin-2在烟曲霉菌感染中的作用及机制。方法用紫外线灭活的烟曲霉菌膨胀态分生孢子刺激野生型(wild type,WT)和Dectin-2缺失(Clec4n-/-)小鼠骨髓来源的巨噬细胞(bone marrow derived macrophages,BMDMs)。刺激20 min及40 min后,利用Western印迹法检测BMDMs中脾酪氨酸激酶(spleen tyrosine kinase,Syk)和核转录因子κB抑制因子(inhibitor-κB,IκBα)的磷酸化水平。刺激16 h后,利用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测BMDMs细胞上清液中IL-6、TNF-α和IL-12p40的水平。另一方面,利用压舌感染的方法在WT和Clec4n-/-小鼠中构建烟曲霉菌肺部感染模型。感染2 d后,统计小鼠整个肺脏荷菌量,并检测肺脏匀浆液中IL-6和IL-12p40水平。结果体外试验提示,烟曲霉菌刺激后,Dectin-2缺失的BMDMs中Syk和IκBα的磷酸化水平及IL-6、TNF-α和IL-12p40水平显著下降(P<0.05)。体内试验发现,烟曲霉菌感染后,Dectin-2缺失小鼠中肺脏荷菌量显著升高(P<0.05),肺脏匀浆液内IL-6、IL-12p40水平显著降低(P<0.05)。结论C型凝集素受体Dectin-2激活烟曲霉菌诱导的NF-κB信号通路并介导促炎细胞因子的产生,可在小鼠肺烟曲霉菌病动物模型中发挥保护性作用。Objective To investigate the role and mechanism of C-type lectin receptor Dectin-2 in Aspergillus fumigatus infection.Methods Bone marrow-derived macrophages(BMDMs)from wild type(WT)and Dectin-2-deficient(Clec4n-/-)mice were stimulated with UV-inactivated Aspergillus fumigatus swollen conidia.The phosphorylation levels of spleen tyrosine kinase(Syk)and inhibitor-κB(IκBα)in BMDMs of WT and Clec4n-/-mice were detected by Western blot after stimulation for 20 and 40 min.The levels of IL-6,TNF-αand IL-12p40 in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA)after stimulation for 16 h.In addition,WT and Clec4n-/-mice were challenged intratracheally with Aspergillus fumigatus conidia for 2 d.The fungal burden of the whole lung was checked,and the levels of IL-6 and IL-12p40 in the lung homogenate were examined in the infected mice.Results In vitro,after stimulation by Aspergillus fumigatus,BMDMs with Dectin-2 deficiency significantly reduced the production of IL-6,TNF-αand IL-12p40 as well as the phosphorylation levels of Syk and IκBαin BMDMs(P<0.05).In vivo,Clec4n-/-mice exhibited significantly heavier fungal burden,while decreased pro-inflammatory cytokine production in lung homogenate compared with WT mice(P<0.05).Conclusion C-type lectin receptor Dectin-2 activates Aspergillus fumigatus induced NF-κB signaling pathway and mediates pro-inflammatory cytokine production,which plays a protective role in defense against pulmonary Aspergillus fumigatus infection.

关 键 词：Dectin-2 C型凝集素受体 小鼠 肺曲霉菌病 烟曲霉菌 

分 类 号：R392[医药卫生—免疫学]