人外周血γδT细胞激活后IL-2和IFN-γ表达的特点

    Expression patterns of IL-2 and IFN-γof activated γδT cells from human peripheral blood

    • 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6~8h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CD28mAb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;MtbAg能诱导γδT细胞表达IL-2,但过程较慢。

       

      Abstract: Objective: To investigate the characteristics of expression of Th1 cytokines interleukin-2(IL-2) and interferon-γ(IFN-γ) of human peripheral γδT cells stimulated with different stimuli.Methods: Human peripheral blood mononuclear cells(PBMCs) were stimulated with phorbol 12-myristate 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-γ 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 γδ 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 cells were not significantly increased.Meanwhile the percentages of IFN-γ-secreting γδ and αβT cells 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 cells among PBMCs default produced of IFN-γ while αβT cells default 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.

       

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