摘要
目的 研究甘露聚糖结合凝集素（mannan-binding lectin,MBL）对辅助性T细胞17（T helper cell 17,Th17）诱导分化的影响.方法 通过MACS分选器从人新鲜脐带血液中分选获得CD4＋T细胞,对照组用anti-CD3 McAb和anti-CD28 McAb活化CD4＋T细胞并加入适当浓度的IL-6和TGF-β1作为诱导剂,在对照组的基础上加入不同浓度的MBL作为实验组,细胞培养72 h后,利用流式细胞术（FACS）检测各组中Th17细胞比例变化情况;Q-PCR分别检测未加MBL对照组和不同浓度MBL实验组Th17细胞核转录因子维甲酸孤儿受体-γt（retinoid-related orphan receptor-γ-t,RORγt）mRNA表达水平;ELISA检测各组细胞上清液中IL-17A的含量;FACS检测Th17细胞在MBL-/-小鼠和WT小鼠中的变化.结果 FACS分析表明,经体外72 h培养,与对照组相比,MBL能够显著降低Th17细胞的比例;Q-PCR结果显示,MBL能够显著降低RORγt mRNA的表达;ELISA结果显示,MBL能够显著降低IL-17A的表达水平;动物实验结果表明,与WT小鼠相比,MBL-/-小鼠CD4＋RORγt＋Th17细胞比例显著上升.结论 MBL能够抑制CD4＋T细胞向Th17细胞诱导分化.
Objective To investigate the effects of mannan-binding lectin （ MBL） on the differen-tiation of Th17 cells （T helper cell 17, Th17）. Methods CD4＋T cells were separated in vitro from fresh human cord blood by MACS （ magnetic-activated cell sorting ） separator. Anti-CD3 monoclonal antibody （ McAb） and anti-CD28 McAb were used to stimulate CD4＋T cells with IL-6 and TGF-β1 as inducers. Then, these cells were treated with （ MBL group） or without （ control group） different concentrations of MBL. Percentages of Th17 cells in different groups were detected by fluorescence-activated cell sorting（ FACS） after 72 hours of culturing. Quantitative real-time PCR （ Q-PCR） was used to analyze the expres-sion of RORγt （ retinoid-related orphan receptor-γ-t） at mRNA level in both control and MBL groups. En-zyme-linked immunosorbent assay （ELISA） was performed to detect the levels of IL-17A in supernatants of cell culture from different groups. FACS was used to detect the percentages of Th17 cells in MBL-/-and wild type （ WT） mice. Results MBL could significantly reduce the percentage of Th17 cells after 72 hours of culturing as compared with the control group. Moreover, MBL could significantly down-regulate the expres-sion of RORγt at mRNA level and decrease the expression of IL-17A. Results of animal experiments showed that the percentages of CD4＋RORγt＋Th17 cells in MBL-/- mice were significantly higher than those in WT mice. Conclusion MBL can inhibit the differentiation of CD4＋T cells to Th17 cells, which is induced by IL-6 and TGF-β1.
作者
陈晨
张卫斌
王艳
杨琳
马俊维
王帅
王凡平
王明永
Chen Chen Zhang Weibin Wang Yan Yang Lin Ma Junwei Wang Shuai Wang Fanping Wang Mingyong(School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China Henan Clinical Medicine （Clinical Laboratory Diagnostics） Gradu- ate Student Innovation Practice Base, Xinxiang Ya Shijie Medical Laboratory, Xinxiang 453003, China Shangqiu Medical College, Shangqiu 476100, China the First People＇s Hospital of Xinxiang 453003, China Department of Obstetrics and Gynecology, the Third Affili- ated Hospital of Xinxiang Medical University, Xinxiang 453003, China Department of Human Parasitology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, China Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang 453003, China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2017年第9期666-673,共8页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目（31100645,81373120）
教育部新世纪优秀人才支持计划（NCET-13-0990）
国家级大学生创新创业训练计划项目（201310472041,201310472033）
新乡医学院研究生科研创新支持计划项目（YJSCX201607Z）.
