摘要
目的探讨树突状细胞经60Co-γ射线照射后激活调节性T淋巴细胞(regulatory T cells,Treg)的能力,以及在Treg诱导的上皮间充质转化(epithelial-mesenchymal transition,EMT)中的作用。方法免疫磁珠分选的小鼠CD4+CD25-常规T细胞(conventional T cells,Tcon)与60Co-γ线照射(6 Gy)后的树突状细胞共培养,72 h后采用流式术检测CD4+CD25+Treg细胞的比例。通过磁珠分选后的Treg与小鼠肺上皮细胞系MLE-12共培养72 h后取MLE-12细胞,免疫荧光和Western印迹技术检测上皮标志蛋白E-钙黏蛋白(E-cadherin,E-cad)和细胞表面蛋白C(prosurfactant protein C,Spc)以及间充质标志蛋白N-钙黏蛋白(N-cadherin,N-cad)和波形蛋白(vimentin)的表达。结果免疫磁珠法分选小鼠脾Treg比例>90%,效果良好;树突状细胞照后抗原提呈功能增强,能显著诱导Tcon分化为Treg; Western印迹和免疫荧光实验证明,Treg能明显促进上皮标志E-cad和Spc表达下调,间充质标志N-cad和vimentin表达升高。结论树突状细胞照射后能显著诱导Treg分化,而Treg能明显诱导肺上皮细胞发生EMT。
Objective To investigate the ability of dendritic cells to activate regulatory T cells( Tregs) after60 Co-γ ray irradiation,and epithelial-mesenchymal transition( EMT) induced by Tregs. Methods Immunomagnetic beads were sorted into mouse CD4+CD25-conventional T cells( Tcons) and co-cultured with60 Co-γ-ray 6 Gy dosed dendritic cells.Flow cytometry was performed after 72 hours of co-culture to detect the proportion of CD4+CD25+Tregs cells. The Tregs after magnetic bead sorting were co-cultured with the mouse lung epithelial cell line MLE-12. After 72 hours of co-culture,MLE-12 cells were detected by immunofluorescence and Western blotting to detect the expression of epithelial marker proteins and interstitial markers. Results The percentage of spleen Tregs cells in the immunomagnetic beads method exceeded 90%,and the effect was good. The antigen-presenting function of dendritic cells was enhanced,which could significantly induce Tcons to differentiate into Tregs cells. Western blotting and immunofluorescence assays simultaneously proved that Tregs could significantly promote EMT in lung epithelial cells. Conclusion Dendritic cells can significantly induce the differentiation of Tregs after irradiation,and Tregs can induce EMT in lung epithelial cells.
作者
孙泽文
梁鑫
郗停停
贺琦多
杨陟华
王易龙
顾永清
潘秀颉
朱茂祥
SUN Ze-wen;LIANG Xin;X;HE Qi-duo;YANG Zhi-hua;WANG Yi-long;GU Yong-qing;PAN Xiu-jie;ZHU Mao-xiang(Graduate School,Guangdong Pharmaceutical University,Guangzhou 510000,China;Institute of Radiation Medicine, Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;General Hospital of PLA, Beijing 100853,China)
出处
《军事医学》
CAS
CSCD
北大核心
2018年第7期502-506,共5页
Military Medical Sciences
基金
国家自然科学基金资助项目(81472910
81673095)
北京市重点实验室资助项目(Z161100005016032)
关键词
辐射
电离
抗原提呈
T淋巴细胞
调节性
上皮间充质转化
钙黏着糖蛋白
肺纤维化
radiation,ionizing
antigen presentation
T-lymphocytes,regulatory
epithelial-mesenchymal transition
cadherins
pulmonary fibrosis
