摘要
目的:探讨人外周血γδT细胞在不同刺激剂激活后产生Th1型细胞因子IL-2和IFN-γ的不同特点.方法:健康成人外周血单个核细胞(PBMC)用佛波醇酯(PMA)、离子霉素(ionomycin)和抗CD28单抗(mAb)以不同组合刺激6～8 h后;用流式细胞术胞内细胞因子检测法测定γδT细胞和αβT细胞IL-2和IFN-γ的表达;或用结核杆菌抗原(Mtb-Ag)刺激γδT细胞加IL-2扩增3～8天后检测γδT细胞IL-2的表达.结果:PBMC经PMA+ionomycin刺激后,与αβT细胞中产生IL-2细胞(14.0%)相比,γδT细胞中极少数细胞(1.1%)表达IL-2;但分泌IFN-γ细胞在γδT细胞中(56.1%)明显多于αβT细胞(8.5%).如加抗CD28 mAb联合刺激后,IL-2产生细胞在αβT细胞中明显增加,但在γδT细胞中未见增加;而IFN-γ表达细胞在两类细胞中均有明显增加;PBMC用Mtb-Ag激活后扩增3天后在γδT细胞中开始检测到IL-2表达,第5～6天时达高峰,第8天时逐渐下降.结论:多克隆短时刺激对PBMC中γδT细胞表达IFN-γ能力强于αβT细胞,但产生IL-2很少;Mtb-Ag能诱导γδT细胞表达IL-2,但过程较慢.
Objective： To investigate the characteristics of expression of Thl eytokines interleukin-2 （IL-2） and interferon-γ（IFN-γ） of human peripheral γδT cells stimulated with different stimuli. Methods： Human peripheral blo：xt mononuclear cells（PBMCs） were stimulated with phorbol 12-myrlstate 13-acetate（PMA） and ionomycin, combined with or without anti-CD28 mAb for 6 to 8 h; or stimulated with Mtb-Ag and cultured in IL-2 containing medium for 3 to 8 days,the intracellular cytokines IL-2 and IFN-δ in γδ and αβT cells were detected by flow cytometry. Results：The percentages of IL-2 secreting cells of γδT cells among PBMCs stimulated with PMA＋ ionomycin were very low （1.1% ） ;compared with that of αβT cells（14.0% ）. The percentages of IFN-γ-secreting cells of γ9 and αβT cells among PBMCs stimulated with stimuli as above were 56.1%, and 8.5 %, respectively. Upon the stimulation of PBMCs with PMA ＋ Ionomycin combined with anti-CD28mAb, the percentages of IL-2 secreting αβT cells increase to 31%, but IL-2 secreting γδT ceils were not significantly increased. Meanwhile the percentages of IFN-γ-secreting γ9 and αβT ceils increase to 77.4 % and 15.4%, respectively. After PBMCs stimulated with Mtb-Ag and cultured in IL-2 containing medium, the percentage of IL-2 producing cells γδT cells were detected only on day 3 and later,increased markedly to a peak on day 6,and thereafter declining to a lower level. Conclusions：Stimulated with PMA ＋ ionomycin combined with or without anti-CD28mAb, the human γδT ceils among PBMCs default produced of IFN-γ while affF cells ddault produced of IL-2. Activated with Mtb-Ag and cultured in presence of IL-2 from 3 to 6 days, γδT cells were also able to produce IL-2.
出处
《蚌埠医学院学报》
CAS
2005年第5期377-380,共4页
Journal of Bengbu Medical College
基金
国家自然科学基金资助项目(30070721)
安徽省高等学校省级重点实验室重点科研项目(2004sys008)
