摘要
目的：研究过表达白细胞介素17受体样分子的人脐带间充质干细胞（IL-17RLM-hUCMSCs）对三硝基苯磺酸（TNBS）诱导的结肠炎小鼠脾脏淋巴细胞的免疫调节作用,为炎症性肠病的干细胞治疗提供优化的种子细胞。方法：体外分离培养hUCMSCs,利用慢病毒载体向干细胞内转入IL-17RLM基因,构建IL-17RLM-hUCMSCs。采用TNBS诱导小鼠实验性结肠炎模型,无菌取炎症小鼠脾脏制备淋巴细胞悬液,在刀豆蛋白A（Con A）刺激下将淋巴细胞分别与不同浓度的IL-17RLM-hUCMSCs及hUCMSCs共培养,72 h后以淋巴细胞＋Con A为阳性对照,CCK8法及CFSE标记法检测淋巴细胞的增殖情况;同时用流式细胞术检测T淋巴细胞亚群（Th1、Th2、Th17及Treg）比例的改变。结果：hUCMSCs及IL-17RLM-hUCMSCs均对ConA刺激下的淋巴细胞增殖有抑制作用（P〈0.05）;当MSCs/淋巴细胞为1∶1~1∶10时,MSCs对淋巴细胞增殖的抑制作用呈现浓度依赖性。在有效浓度范围内,IL-17RLM-hUCMSCs较h UCMSCs抑制作用更强（P〈0.05）。hUCMSCs及IL-17RLM-h UCMSCs均能够下调结肠炎小鼠脾脏淋巴细胞的Th1和Th17细胞亚群比例,上调Treg细胞亚群比例,但IL-17RLM-h UCMSCs对Th17细胞亚群的抑制作用更显著（P〈0.05）。结论：IL-17RLM-hUCMSCs呈浓度依赖性地抑制TNBS诱导的结肠炎小鼠脾脏淋巴细胞增殖,且该作用优于h UCMSCs。同时,IL-17RLM-hUCMSCs可调节T细胞亚群的免疫平衡,且抑制Th17细胞亚群作用强于hUCMSCs。
AIM： To investigate the immunomodulatory effect of human umbilical cord mesenchymal stem cells with interleukin-17 receptor-like molecule overexpression（ IL-17RLM-hUCMSCs） on the spleen lymphocytes from the mice with trinitrobenzene sulfonic acid（ TNBS）-induced colitis for providing optimal seed cells to treat inflammatory bowel disease. METHODS： Mesenchymal stem cells were isolated from human umbilical cord. The IL-17 RLM gene was transferred into mesenchymal stem cells by lentivirus vector to establish IL-17RLM-hUCMSCs. The experimental colitis mice were induced by TNBS enema,and the spleen lymphocyte suspension was isolated. The lymphocytes were co-cultured with different concentrations of IL-17RLM-hUCMSCs and hUCMSCs under the stimulation of concanavalin A（ Con A） for 72 h. The proliferation of lymphocytes was detected by the methods of CCK8 assay and CFSE labeling with lymphocytes ＋ Con A as positive control. The changes of lymphocyte subsets（ Th1,Th2,Th17 and Treg） were examined by flow cytometry. RESULTS： Both hUCMSCs and IL-17RLM-h UCMSCs inhibited T-cell proliferation in vitro co-culture system（ P〈0. 05）.When the ratios of MSCs to lymphocytes ranged from 1∶ 1 to 1∶ 10,the inhibitory rates were in a dose-dependent manner.IL-17RLM-h UCMSCs showed higher inhibitory rate than hUCMSCs within the effective concentration range（ P〈0. 05）.Both h UCMSCs and IL-17RLM-hUCMSCs reduced the proportions of Th1 and Th17 cell subsets and increased Treg cell population of spleen lymphocytes from TNBS-induced colitis mice,and IL-17RLM-hUCMSCs showed a stronger inhibitory effect on Th17 cell subset（ P〈0. 05）. CONCLUSION： IL-17RLM-hUCMSCs remarkably inhibit the proliferation of spleen lymphocytes from TNBS-induced colitis mice in a dose-dependent manner. Meanwhile,they regulate immune balance of T cells and have stronger inhibitory effect on Th17 subset.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第6期961-970,共10页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81370498)
关键词
脐带间充质干细胞
白细胞介素17受体样分子
淋巴细胞
免疫调节
Umbilical cord-derived mesenchymal stem cells
Interleukin-17 receptor-like molecule
Lymphocytes
Immunomodulation
