环孢素对正常人PBMCs产生IFN-γ的抑制作用

Inhibitory Effect of Ciclosporine on Production of IFN-γ by Human PBMCs

【目的】探讨在不同刺激条件下,环孢素(ciclosporine,CsA)对正常人外周血单个核细胞(PBMCs)γ-干扰素(interferon-gamma,IFN-γ)的产生,以及对细胞亚群的影响。【方法】PBMCs在抗CD3单克隆抗体(anti-CD3)、抗CD3和抗CD28(anti-CD28)单克隆抗体、IL-12(Th1细胞分化因子)刺激的情况下或在混合淋巴细胞反应中,观察CsA对IFN-γ产生和Th1细胞分化的影响。同时使用流式细胞仪,在单个细胞水平上评价CsA对T淋巴细胞表面活化分子CD25及细胞内细胞因子IFN-γ和IL-2产生的影响。【结果】CsA对由不同条件诱导产生的IFN-γ,均表现为剂量依赖性抑制关系。此外,CsA抑制IL-12诱导的Th1分化。进一步研究结果表明,CsA可抑制CD4+和CD8+T细胞IFN-γ的表达。【结论】CsA剂量依赖性抑制T淋巴细胞的活化,抑制CD4+、CD8+T淋巴细胞产生IFN-γ和Th1细胞的分化,其抑制机理可能与细胞的活化有关。

[Objective] To evaluate the effect of ciclosporine (CsA) on the production of IFN-γ by peripheral blood mononuclear cells (PBMCs) and on the cell subsets following stimulation under different culture condition.[Methods]PBMCs were prepared and cultured either with anti-CD3 alone, anti-CD3 plus anti-CD28, IL-12 or mixed leukocyte culture (MLC) in the presence or absence of CsA at different concentration. After stimulation for 3 or 5 days, the level of IFN-γ in the culture supernatant was assayed by ELISA. In addition, the cultured cells were harvested, restimulated and stained for the expression of intracellular cytokines. The frequency of IFN-γ-producing and IL-2-producing cells by CD4+ and CD8+ T cells was examined at a single cell level by flow cytometric analysis.[Results]After stimulation at different culture conditions, T cells produced high levels of IFN-γ. CsA inhibited INF-γ production in a dose dependent manner. The inhibitory effects of CsA on IL-12-induced differentiation of Th1 cells and IL-12 responsiveness were observed as well. Further analysis indicated that CsA suppressed IFN-γ expression by both CD4+ and CD8+ T cells.[Conclusion] CsA inhibits the production of IFN-γ by both CD4+ and CD8+ T cells and the differentiation of Th1 cells via a mechanism of suppression of early T cell activation.