中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
9期
792-797
,共6页
栾弘%张茜%王艻%张妙%陈艳%许小利%李杏爱%刘丽丽%袁劲%吕永曼
欒弘%張茜%王艻%張妙%陳豔%許小利%李杏愛%劉麗麗%袁勁%呂永曼
란홍%장천%왕륵%장묘%진염%허소리%리행애%류려려%원경%려영만
Galectin-9%Th1%Th2%Th17
Galectin-9%Th1%Th2%Th17
Galectin-9%Th1%Th2%Th17
Galectin-9%Th1%Th2%Th17
目的 研究Galectin-9对活化的CD4+T细胞的免疫调节作用,并进一步探讨其作用机制.方法 获取野生型C57BL/6小鼠淋巴细胞,利用MACS分选CD4+ na(i)ve T细胞,给予anti-CD3 (2.5 μg/ml)抗体、anti-CD28(5μg/ml)抗体和IL-2(100 ng/ml)刺激,培养3d.将活化的CD4+T细胞分为3组:正常对照组、Galectin-9组和Galectin-9+α-乳糖组.利用CFSE检测T细胞增殖情况,并动态观察细胞形态变化；检测CD4+ CD69+T细胞、Th1、Th2和Th17细胞的比例；并利用ELISA检测淋巴细胞分泌IFN-γ 、IL-4、IL-10、IL-12、IL-17A和TGF-β1等细胞因子的水平；利用Western blot检测T-bet、GATA-3和ROR-yt等T细胞分化调控蛋白的变化.结果 与正常对照组和Galectin-9+α-乳糖组相比,Galectin-9组于2h时开始出现细胞形态改变,同时CD4+ CD69+T细胞、Th1和Th17细胞表达减少(P＜0.05),但Th2细胞无明显变化；培养上清液中IFN-γ、IL-12、IL-17A和TGF-β1的表达水平降低(P＜0.05),而IL-4和IL-10等Th2型细胞因子水平无明显变化；同时T-bet和ROR-γt的表达水平减少(P＜0.05).结论 Galectin-9抑制活化CD4+T细胞的Th1和Th17型免疫应答,而对Th2型免疫应答无影响,免疫调节的机制可能与其在转录水平影响Th1和Th17特定转录因子的表达有关.
目的 研究Galectin-9對活化的CD4+T細胞的免疫調節作用,併進一步探討其作用機製.方法 穫取野生型C57BL/6小鼠淋巴細胞,利用MACS分選CD4+ na(i)ve T細胞,給予anti-CD3 (2.5 μg/ml)抗體、anti-CD28(5μg/ml)抗體和IL-2(100 ng/ml)刺激,培養3d.將活化的CD4+T細胞分為3組:正常對照組、Galectin-9組和Galectin-9+α-乳糖組.利用CFSE檢測T細胞增殖情況,併動態觀察細胞形態變化；檢測CD4+ CD69+T細胞、Th1、Th2和Th17細胞的比例；併利用ELISA檢測淋巴細胞分泌IFN-γ 、IL-4、IL-10、IL-12、IL-17A和TGF-β1等細胞因子的水平；利用Western blot檢測T-bet、GATA-3和ROR-yt等T細胞分化調控蛋白的變化.結果 與正常對照組和Galectin-9+α-乳糖組相比,Galectin-9組于2h時開始齣現細胞形態改變,同時CD4+ CD69+T細胞、Th1和Th17細胞錶達減少(P＜0.05),但Th2細胞無明顯變化；培養上清液中IFN-γ、IL-12、IL-17A和TGF-β1的錶達水平降低(P＜0.05),而IL-4和IL-10等Th2型細胞因子水平無明顯變化；同時T-bet和ROR-γt的錶達水平減少(P＜0.05).結論 Galectin-9抑製活化CD4+T細胞的Th1和Th17型免疫應答,而對Th2型免疫應答無影響,免疫調節的機製可能與其在轉錄水平影響Th1和Th17特定轉錄因子的錶達有關.
목적 연구Galectin-9대활화적CD4+T세포적면역조절작용,병진일보탐토기작용궤제.방법 획취야생형C57BL/6소서림파세포,이용MACS분선CD4+ na(i)ve T세포,급여anti-CD3 (2.5 μg/ml)항체、anti-CD28(5μg/ml)항체화IL-2(100 ng/ml)자격,배양3d.장활화적CD4+T세포분위3조:정상대조조、Galectin-9조화Galectin-9+α-유당조.이용CFSE검측T세포증식정황,병동태관찰세포형태변화；검측CD4+ CD69+T세포、Th1、Th2화Th17세포적비례；병이용ELISA검측림파세포분비IFN-γ 、IL-4、IL-10、IL-12、IL-17A화TGF-β1등세포인자적수평；이용Western blot검측T-bet、GATA-3화ROR-yt등T세포분화조공단백적변화.결과 여정상대조조화Galectin-9+α-유당조상비,Galectin-9조우2h시개시출현세포형태개변,동시CD4+ CD69+T세포、Th1화Th17세포표체감소(P＜0.05),단Th2세포무명현변화；배양상청액중IFN-γ、IL-12、IL-17A화TGF-β1적표체수평강저(P＜0.05),이IL-4화IL-10등Th2형세포인자수평무명현변화；동시T-bet화ROR-γt적표체수평감소(P＜0.05).결론 Galectin-9억제활화CD4+T세포적Th1화Th17형면역응답,이대Th2형면역응답무영향,면역조절적궤제가능여기재전록수평영향Th1화Th17특정전록인자적표체유관.
Objective To study the immune regulation of Galectin-9 on the active CD4+T cells and demonstrate the mechanisms.Methods Lymphocytes were harvested from wild-type C57BL/6 mouse,from which na(i)ve CD4+T cells were separated via MACS and then stimulated with anti-CD3 antibody(Ab) (2.5 μg/ml),anti-CD28 Ab(5 μg/ml) and IL-2(100 ng/ml) for 3 days.The active CD4+T cells were divided into 3 groups:Control group,Galectin-9 group and Galectin-9+α-lactose group.We detected the cell proliferation level by CFSE fluorescence intensity and then dynamically observed the cell morphological changes.The proportion of CD4+CD69+T cell,Th1,Th2 and Th17 cell was valued; Meanwhile,ELISA was used to detect the cytokine levels of IFN-γ,IL-4,IL-10,IL-12,IL-17A and TGF-β1 secreted by lymphocytes.Also Western blot was used to observe the changes of T cell differentiation regulatory protein such as T-bet,GATA-3 and ROR-γt.Results Compared with control group and Galectin-9+α-lactose group,in Galectin-9 group,the cell morphology began to change at 2 h.Moreover the proportions of CD4+CD69+ T cell,Th1 and Th17 cells decreased (P＜0.05),but no significant differences in Th2 cells.The level of IFN-γ,IL-12,IL-17A and TGF-β1 from the supernatant decreased (P＜0.05),while Th2-type cytokines IL-4 and IL-1O did not change.In addition,the expressions of T-bet and ROR-γt were significantly down-regulated (P＜0.05).Conclusion Galectin-9 inhibited Th1 and Th17-type immune response,while had no effect on Th2-type immune response.The mechanism of the immune regulation may be related to affect the expression of Th1 and Th17 specific transcription factors at transcription level.
