天津医科大学学报
天津醫科大學學報
천진의과대학학보
Journal of Tianjin Medical University
2015年
5期
393-396
,共4页
诱导性调节性T细胞%Foxp3%TGF-β%免疫抑制
誘導性調節性T細胞%Foxp3%TGF-β%免疫抑製
유도성조절성T세포%Foxp3%TGF-β%면역억제
induced regulatory T cells%Foxp3%TGF-β%immunosuppression
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. 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IL-10、TGF-β分泌以及胞内因子IL-2、IL-4、IL-17、干扰素-γ(IFN-γ)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-γ. 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.
目的:製備誘導性調節性T淋巴細胞(iTreg),探討iTreg的免疫抑製功能及其作用機製. 方法:免疫磁珠分選穫取CD4+CD25-T淋巴細胞,TGF-β誘導法穫取iTreg,流式細胞術和實時定量PCR方法檢測iTreg的得率及其扠頭狀/翅膀狀螺鏇轉錄因子(Foxp3)mRNA水平,活性染料羧基熒光素乙酰乙痠琥珀酰亞胺酯(CFSE)染色和流式細胞術方法觀察Treg細胞對CD4+T淋巴細胞體外增殖能力的影響.利用ELISA和流式細胞術方法分彆檢測iTreg的IL-10、TGF-β分泌以及胞內因子IL-2、IL-4、IL-17、榦擾素-γ(IFN-γ)水平. 結果:TGF-β誘導法iTreg得率可達42.10%,TGF-β誘導後穫得的iTreg Foxp3 mRNA水平明顯升高. 穫得的iTreg可以抑製自體CD4+T淋巴細胞增殖,IL-10和TGF-β分泌水平較原始CD4+T細胞上升(P<0.05),但幾乎不分泌IL-2、IL-4、IL-17、IFN-γ. 結論:利用TGF-β誘導法製備穫得的iTreg具有明顯的免疫抑製功能,其髮揮免疫抑製功能過程可能均有IL-10和TGF-β的參與.
목적:제비유도성조절성T림파세포(iTreg),탐토iTreg적면역억제공능급기작용궤제. 방법:면역자주분선획취CD4+CD25-T림파세포,TGF-β유도법획취iTreg,류식세포술화실시정량PCR방법검측iTreg적득솔급기차두상/시방상라선전록인자(Foxp3)mRNA수평,활성염료최기형광소을선을산호박선아알지(CFSE)염색화류식세포술방법관찰Treg세포대CD4+T림파세포체외증식능력적영향.이용ELISA화류식세포술방법분별검측iTreg적IL-10、TGF-β분비이급포내인자IL-2、IL-4、IL-17、간우소-γ(IFN-γ)수평. 결과:TGF-β유도법iTreg득솔가체42.10%,TGF-β유도후획득적iTreg Foxp3 mRNA수평명현승고. 획득적iTreg가이억제자체CD4+T림파세포증식,IL-10화TGF-β분비수평교원시CD4+T세포상승(P<0.05),단궤호불분비IL-2、IL-4、IL-17、IFN-γ. 결론:이용TGF-β유도법제비획득적iTreg구유명현적면역억제공능,기발휘면역억제공능과정가능균유IL-10화TGF-β적삼여.
Objective:To establish TGF-βinduced method to obtain induced regulatory T cells (iTreg), and to study the immunosuppression function and mechanism of iTreg. Methods:CD4+CD25-T were obtained by MACS, after which the iTreg cells were obtained by TGF-βinduced 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-βsecretion levels and intracellular cytokine levels of IL-2, IL-4, IL-17 and IFN-γ. Results:The yield rate of iTreg cells was increased to 42.10%using by TGF-βinduced 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-βsecretion were significantly inhibited by iTreg cells versus non-Treg cells (P<0.05). IL-2, IL-4, IL-17 and IFN-γwere hardly secreted by iTreg cells. Conclusion:TGF-βinduction can be successfully used to obtain iTreg. IL-10 and TGF-βmay be both involved in iTreg immunosuppression function.
