与脐血多能干细胞共培养的小鼠淋巴细胞对AD小鼠的免疫调节作用

基础医学与临床 ›› 2017, Vol. 37 ›› Issue (8): 1072-1076.

与脐血多能干细胞共培养的小鼠淋巴细胞对AD小鼠的免疫调节作用

贺毅¹,李恒²,张凤³,唐小三¹,张贵涛³,李晓红³

1. 山东大学医学院
2. 山东大学附属济南市中心医院神经内科
3. 山东大学附属济南市中心医院

收稿日期:2016-06-06 修回日期:2016-09-25 出版日期:2017-08-05 发布日期:2017-07-17
通讯作者: 李晓红 E-mail:Xiaohong-Li@sdu.edu.cn
基金资助:
国家自然科学基金面上项目;济南市科技发展国际合作项目

Immune modulation by mouse lymphocytes co-cultured with cord blood-derived multipotent stem cells in Alzheimer‘s disease model mice

Received:2016-06-06 Revised:2016-09-25 Online:2017-08-05 Published:2017-07-17
Supported by:
; Jinan Municipal Science and Technology Development Program

摘要/Abstract

摘要： 目的探讨与脐血多能干细胞(CB-SCs）共培养的淋巴细胞对阿尔兹海默病(AD)小鼠的免疫调节作用及治疗潜能。方法从人脐血中分离培养CB-SCs；从小鼠脾脏分离淋巴细胞；淋巴细胞与CB-SCs共同培养72h或淋巴细胞单独培养；将AD小鼠随机分为实验组与对照组，实验组尾静脉注射与CB-SCs 共培养的淋巴细胞，对照组尾静脉注射单独培养的淋巴细胞。注射结束后进行行为学实验,流式细胞学检测外周血淋巴细胞中调节性T细胞（Tregs）比例, ELISA检测小鼠外周血血浆中TNF-α和IL-10的表达，PCR检测AD小鼠脑内炎性因子的表达，免疫荧光染色检测AD小鼠脑内淀粉样斑块（β-amyloid, Aβ）含量。结果1）实验组小鼠的空间学习记忆能力得到改善；2）实验组小鼠脑内的Aβ沉积量低于对照组；3）实验组外周血淋巴细胞中Tregs百分比及血浆中的抗炎因子IL-10浓度高于对照组（P ＜0.05），而血浆中的促炎因子TNF-α浓度低于对照组（P ＜0.001）；4）实验组小鼠脑内IL-10的mRNA表达量高于对照组（P ＜0.05），而TNF-α的mRNA表达量低于对照组（P ＜0.05）。结论与脐血多能干细胞共培养的淋巴细胞对AD小鼠具有免疫治疗作用，主要通过增加Tregs细胞的比例并增强其抗炎功能来发挥作用。

关键词: 阿尔兹海默病（AD）, 脐血多能干细胞(CB-SCs), 淋巴细胞, 免疫调节

Abstract: Objective To investigate the immune modulation effect of lymphocytes co-cultured with human cord blood derived-multipotent stem cells(CB-SCs) and further to explore their therapeutic potential for Alzheimer’s Disease (AD) model mice. Methods CB-SCs were isolated from human cord blood. Lymphocytes were isolated from the spleens of AD model mice. The lymphocytes were co-cultured with CB-SCs or cultured alone for 72 h. AD model mice were divided into experimental group and control group randomly, and then the experimental group mice were administrated with lymphocytes co-cultured with CB-SCs and control group were administrated with lymphocytes cultured alone by caudal vein injection. Then, the behavior experiment was carried out. The CD4+CD25+Foxp3+Tregs were detected by flow cytometry. The protein expression of TNF-αand IL-10 in peripheral blood was detected by ELISA, The mRNA expression of TNF-α and IL-10 in brain tissue was detected by PCR. The amyloid-β（Aβ）plaques were detected by immunohistochemistry. Results 1) There is spatial learning and memory improvement on experimental group. 2) The Aβ plaques of experimental group were decreased compared to the control group. 3) The percentage of CD4+CD25+Foxp3+Tregs in peripheral blood and IL-10 level in plasma were higher in experimental group compared to those in the control group（P ＜0.05）. The pro-inflammatory cytokines, TNF-α level in plasma of experimental group was lower than that in the control group（P ＜0.001）. 4) The mRNA expression of IL-10 in brain was higher in experimental group compared to those in the control group（P ＜0.05）, and the mRNA expression of TNF-α of experimental group was lower than that in the control group（P ＜0.05）. Conclusion The lymphocytes co-cultured with CB-SCs have immunotherapeutic effect on AD model mice, which is mainly displayed with increasing proportion of Tregs and enhanced anti-inflammatory function of Tregs．

Key words: Alzheimer’s Disease（AD）, cord blood-derived multipotent stem cells（CB-SCs）, lymphocytes, immune modulation

中图分类号:

R741.05

贺毅李恒张凤唐小三张贵涛李晓红. 与脐血多能干细胞共培养的小鼠淋巴细胞对AD小鼠的免疫调节作用[J]. 基础医学与临床, 2017, 37(8): 1072-1076.

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