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目的研究抗原特异性T细胞活化后,其所表达的Foxp3、CTLA-4和细胞因子的动态变化,为深入了解特异性免疫应答调控奠定基础。方法体外采用树突状细胞(DC)系D2SC/1作为抗原提呈细胞(APC)将OVA323-339直接提呈给OVA特异性TCR转基因小鼠DO11.10来源的CD4~+T细胞。采用CCK-8法测定细胞活化增殖情况,并应用Real time-PCR动态分析细胞因子IL-2、IFN-γ、IL-10、TGF-β,CTLA-4和Foxp3的mRNA表达水平。结果OVA特异性T细胞活化所需的最佳OVA剂量为2μmol/L,T细胞活化后IL-2、IFN-γ、IL-10和TGF-βmRNA表达水平最高峰值分别在8、4、2 h和8 h,Foxp3 mRNA在2 h时有一过性的表达峰,随后在24 h又有所升高,而CTLA-4 mRNA在静止时有较高水平的表达,但活化后迅速下调,其在24 h内无明显变化。结论初步了解了适应性免疫应答中Foxp3、CTLA-4和主要免疫调节相关的细胞因子表达趋势,为深入研究适应性免疫应答调控机制奠定初步基础。
Objective To study the dynamic changes of Foxp3, CTLA-4 and cytokines expressed by antigen-specific T cells in order to further understand the regulation of specific immune responses. Methods OVA323-339 was directly presented to DO11.10-derived CD4 ~ + T cells of OVA-specific TCR transgenic mice using dendritic cell (DC) line D2SC / 1 as antigen presenting cell (APC) in vitro. The activation and proliferation of cells were measured by CCK-8. The mRNA expression of cytokines IL-2, IFN-γ, IL-10, TGF-β, CTLA-4 and Foxp3 were analyzed by Real time- Results The optimal OVA dose required for OVA-specific T cell activation was 2 μmol / L. The highest peak of the expression of IL-2, IFN-γ, IL-10 and TGF-β mRNA after T cell activation was at 8, And 8 h, Foxp3 mRNA had a transient expression peak at 2 h, then increased again at 24 h, whereas CTLA-4 mRNA showed a higher level at rest but rapidly down-regulated after activation No significant change within 24 h. Conclusion The expression of Foxp3, CTLA-4 and the major immunoregulatory cytokines in adaptive immune response are preliminarily understood, which provides a preliminary basis for further study on the regulatory mechanism of adaptive immune response.