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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

21卷

期数:

2015年05期

页码:

393-396

栏目:

基础医学

出版日期:

2015-09-20

文章信息/Info

Title:

Study on iTreg cells obtaining method and its immunosuppression function

文章编号:

1006-8147（2015）05-0393-04

作者:

王 凊¹; 车绪春²

?（1.天津医科大学总医院检验科，天津300052；2. 天津医科大学免疫学系，天津300070）

Author(s):

WANG Qing¹;  CHE Xu-chun²

(1. Department of Clinical Laboratory, General Hospital Tianjin Medical University, Tianjin 300052, China; 2. Department of Immunolgy, Tianjin Medical University, Tianjin 300070, China)

关键词:

?诱导性调节性T细胞' target="_blank" rel="external">">?诱导性调节性T细胞; Foxp3; TGF-β; 免疫抑制

Keywords:

induced regulatory T cells' target="_blank" rel="external">">induced regulatory T cells;  Foxp3;  TGF-β;  immunosuppression

' target="_blank" rel="external">" />

分类号:

R392.12

DOI:

-

文献标志码:

A

摘要:

目的：制备诱导性调节性T淋巴细胞（iTreg），探讨iTreg的免疫抑制功能及其作用机制研究。方法：免疫磁珠分选获取CD4⁺CD25^-T淋巴细胞，TGF-β诱导法获取iTreg，流式细胞术和实时定量PCR方法检测iTreg的得率及其叉头状/翅膀状螺旋转录因子（Foxp3）mRNA水平，活性染料羧基荧光素乙酰乙酸琥珀酰亚胺酯（CFSE）染色和流式细胞术方法观察Treg细胞对CD4⁺T淋巴细胞体外增殖能力的影响。利用ELISA和流式细胞术方法分别检测iTreg的白细胞介素-10（IL-10）、TGF-β分泌，以及胞内因子IL-2、IL-4、IL-17、干扰素-g（IFN-g）水平。结果：TGF-β诱导法iTreg得率可达42.10% ，TGF-β诱导后获得的iTreg Foxp3 mRNA水平明显升高。获得的iTreg可以抑制自体CD4⁺T淋巴细胞增殖，IL-10和TGF-β分泌水平较原始CD4⁺ T细胞上升（P<0.05），但几乎不分泌IL-2、IL-4、IL-17、IFN-g。结论：利用TGF-β诱导法制备获得的iTreg具有明显的免疫抑制功能，其发挥免疫抑制功能过程可能均有IL-10和TGF-β的参与。

Abstract:

Objective: To establish TGF-β induced method to obtain induced regulatory T cells (iTreg), and to study the immunosuppression fuction and mechanism of iTreg . Methods: CD4⁺CD25^-T were obtained by MACS, after which the iTreg cells were obtained by TGF-β induced method. Flow cytometry and PCR were used to detect iTreg yield and Foxp3 mRNA level, CFSE staining and Flow cytometry were applied to investigate the effect of iTreg on proliferation activity of CD4⁺T. ELISA and Flow cytometry were adopted to detect iTreg IL-10 and TGF-β secretion levels and intracellular cytokine levels of IL-2, IL-4, IL-17 and IFN-g. Results: The yield rate of iTreg cells was increased to 42.10% using by TGF-β induced method. The Foxp3 mRNA of the induced iTreg were higher than that of CD4⁺T (P<0.05). CD4⁺T cell proliferation and IL-10, TGF-β secretion were significantly inhibited by iTreg cells versus non-Treg cells (P<0.05). IL-2, IL-4, IL-17 and IFN-g were hardly secreted by iTreg cells. Conclusion: TGF-β induction can be successfully used to obtain iTreg. IL-10 and TGF-β may be both involved in iTreg immunosuppression function.

?

参考文献/References:

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备注/Memo

备注/Memo:

作者简介 王凊（1978-），女，主管技师，硕士，研究方向：免疫调节及抗肿瘤免疫；
通信作者：车绪春， E-mail ：chexuchun@163.com。

常用功能

更新日期/Last Update: 2015-09-21