IL-37b通过抑制树突状细胞相关的免疫应答缓解感染性休克  

IL-37b alleviates septic shock through inhibition of immune responses related to dendritic cells

作　　者：李梦媛[1] 燕正强 韦荣飞 杨星九 朱瑞敏[1] 高苒[1] LI Meng yuan;YAN Zheng qiang;WEI Rong fei;YANG Xing jiu;ZHU Rui min;GAO Ran(Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences (CAMS) & Comparative Medicine Center, Peking Union Medical College (PUMC), Beijing 100021, China;Department of Orthopedics,Dingxi Hospital of Traditional Chinese Medicine,Dingxi 743000,China)

机构地区：[1]中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京100021 [2]定西市中医院骨伤一科,甘肃定西743000

出　　处：《中国比较医学杂志》2017年第11期44-49,共6页Chinese Journal of Comparative Medicine

基　　金：中国医学科学院医学与健康科技创新工程-重大协同创新项目-协同创新团队资助(编号:2016-I2M-3-019)

摘　　要：目的表达IL-37b重组蛋白,研究其在调节机体免疫应答方面的作用机制。方法构建原核表达载体p ET28/IL-37b,诱导表达的IL-37b重组蛋白由Ni^(2+)-NTA凝胶进行纯化。将IL-37b作用于C57BL/6 J小鼠,注射LPS诱导感染性休克,检测小鼠血清中IL-1β、IL-6、IFN-γ的表达水平。分离并培养小鼠树突状细胞,用IL-37b处理,经LPS诱导活化,检测细胞表面CD40、CD80、MHCII等标志物分子表达情况,以及培养上清中IL-1β、IL-10、IL-12、TNF-α的表达水平。分离小鼠CD4^+T细胞,检测IL-37b对LPS诱导的T细胞培养上清中IFN-γ、TNF-α、IL-10的抑制作用。结果IL-37b能够降低感染性休克的小鼠血清中促炎性细胞因子的表达;并抑制小鼠树突状细胞共刺激分子和促炎性细胞因子的表达,以及CD4^+T细胞表达促炎性细胞因子。结论纯化的IL-37b具备生物学活性,能够通过抑制树突状细胞活化及相关的T细胞免疫应答缓解机体感染性休克。ObjectiveTo express the recombinant IL37b protein and to investigate its role in the regulation of immune responses.MethodsThe prokaryotic expression vector pET28/IL37b was constructed.The recombinant IL37b protein was induced to be expressed and was purified using Ni2+NTA gel column.C57BL/6J mice were treated with IL37b and injected with lipopolysaccharide(LPS)to induce septic shock,and the expression levels of IL1β,IL6,IFNγin the serum of the mice were detected.Dendritic cells from bone marrow of the mice were isolated and cultured,and were treated with IL37b.After LPS induced activation,the expression levels of marker molecules such as CD40,CD80and MHCII on the cell surface,and cytokines such as IL1β,IL10,IL12and TNFαin the culture supernatants were detected by flow cytometry.The CD4+T cells from mice were isolated and the inhibitory effects of IL37b on the expression of IFNγ,TNFαand IL10in the culture supernatants of T cells after induction by LPS were detected.ResultsIL37b reduced the expression levels of proinflammatory cytokines in the serum of septic shock mice.IL37b also inhibited the expression of co stimulatory molecules and proinflammatory cytokines of the mouse dendritic cells,and suppress the activation of CD4+T cells in vitro.ConclusionsPurified recombinant IL37b protein has high bioactivity,and can alleviate septic shock in organisms through inhibiting the activation of dendritic cells and related T cell immune responses.

关 键 词：IL37b 纯化 树突状细胞 免疫应答 感染性休克 

分 类 号：R-33[医药卫生]