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
2010年
2期
110-114
,共5页
邱莲女%周永列%陈秀君%胡惟孝
邱蓮女%週永列%陳秀君%鬍惟孝
구련녀%주영렬%진수군%호유효
ZGDHu-1%环孢霉素A%T细胞活化%细胞凋亡%流式细胞术
ZGDHu-1%環孢黴素A%T細胞活化%細胞凋亡%流式細胞術
ZGDHu-1%배포매소A%T세포활화%세포조망%류식세포술
ZGDHu-1%Cyclosporin A%T cells activation%Apoptosis%Flow cytometry
目的 研究四嗪二甲酰胺(ZGDHu-1)对植物血凝素(PHA)刺激的T淋巴细胞体外活化的影响,初步探讨其免疫抑制作用.方法 从健康人外周血分离淋巴细胞,经PHA刺激诱导淋巴细胞活化,ZGDHu-1单独或联合环孢霉素A(CSA)作用24 h、48 h,检测各组T淋巴细胞的增殖、凋亡情况.采用流式细胞仪检测CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+、CD8~+ CD25~+和CD3~+ Fas~+、CD4~+ Fas~+、CD8~+ Fas~+.用Annexin V/PI染色结合流式细胞术,分析淋巴细胞早期凋亡.酶联免疫吸附法(ELISA)检测细胞培养上清IL-2和转化生长因子-β1(TGF-β1).结果 ZGDHu-1能明显降低PHA活化的CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+和CD3~+ Fas~+、CD4~+ Fas~+、Annexin V~+/PI~-,并降低活化T淋巴细胞的IL-2分泌及促进TGF-β1的分泌,联合CSA应用效果更明显.结论 ZGDHu-1能抑制T淋巴细胞活化,降低T淋巴细胞凋亡,和CSA联合具有一定的协同效应.
目的 研究四嗪二甲酰胺(ZGDHu-1)對植物血凝素(PHA)刺激的T淋巴細胞體外活化的影響,初步探討其免疫抑製作用.方法 從健康人外週血分離淋巴細胞,經PHA刺激誘導淋巴細胞活化,ZGDHu-1單獨或聯閤環孢黴素A(CSA)作用24 h、48 h,檢測各組T淋巴細胞的增殖、凋亡情況.採用流式細胞儀檢測CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+、CD8~+ CD25~+和CD3~+ Fas~+、CD4~+ Fas~+、CD8~+ Fas~+.用Annexin V/PI染色結閤流式細胞術,分析淋巴細胞早期凋亡.酶聯免疫吸附法(ELISA)檢測細胞培養上清IL-2和轉化生長因子-β1(TGF-β1).結果 ZGDHu-1能明顯降低PHA活化的CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+和CD3~+ Fas~+、CD4~+ Fas~+、Annexin V~+/PI~-,併降低活化T淋巴細胞的IL-2分泌及促進TGF-β1的分泌,聯閤CSA應用效果更明顯.結論 ZGDHu-1能抑製T淋巴細胞活化,降低T淋巴細胞凋亡,和CSA聯閤具有一定的協同效應.
목적 연구사진이갑선알(ZGDHu-1)대식물혈응소(PHA)자격적T림파세포체외활화적영향,초보탐토기면역억제작용.방법 종건강인외주혈분리림파세포,경PHA자격유도림파세포활화,ZGDHu-1단독혹연합배포매소A(CSA)작용24 h、48 h,검측각조T림파세포적증식、조망정황.채용류식세포의검측CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+、CD8~+ CD25~+화CD3~+ Fas~+、CD4~+ Fas~+、CD8~+ Fas~+.용Annexin V/PI염색결합류식세포술,분석림파세포조기조망.매련면역흡부법(ELISA)검측세포배양상청IL-2화전화생장인자-β1(TGF-β1).결과 ZGDHu-1능명현강저PHA활화적CD3~+ CD69~+、CD3~+ CD25~+、CD4~+ CD25~+화CD3~+ Fas~+、CD4~+ Fas~+、Annexin V~+/PI~-,병강저활화T림파세포적IL-2분비급촉진TGF-β1적분비,연합CSA응용효과경명현.결론 ZGDHu-1능억제T림파세포활화,강저T림파세포조망,화CSA연합구유일정적협동효응.
Objective To investigate the effects of ZGDHu-1 on T lymphocytes activation in vitro to elucidate its immunosuppressive effects. Methods Lymphocytes isolated from healthy persons were stim-ulated with phytohanmagglutinin(PHA) and different experimental groups were set by cocultured for 24 h, 48 h with ZGDHu-1 or with ZGDHu-1 and Cyclosporin A(CSA). To assess the proliferation and apoptosis of T lymphocytes, we detected CD3~+ CD69~+ , CD3~+ CD25~+ , CD4~+ CD25~+ , CD8~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+ , CD8~+ Fas~+ with flow cytometry. The early apoptosis rate of lymphocytes was analyzed with flow cytometry. Culture supernatant IL-2 and TGF-β1 were detected with ELISA. Results ZGDHu-1 decreased PHA activative CD3~+ CD69~+, CD3~+ CD25~+, CD4~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+, Annexin V~+/ PI~- and inhibited IL-2 secretion and promoted TGF-β1 secretion respectively. ZGDHu-1 has synergistic effect with CSA to be more obvious. Conclusion ZGDHu-1 can inhibit T lymphocytes activation and de-creased apeptosis of T lymphocytes. ZGDHu-1 has synergistic effect with CSA to be obvious.
