显性调控下免疫细胞局部微环境的特征

Microenviroment character of immune response under dominant regulation

目的 观察显性调控过程中细胞因子谱系的动态变化及不同刺激强度的影响作用 ,归纳显性调控作用下免疫反应局部微环境的特征。方法 分离外周血单个核细胞 ,采用不同组合刺激 ,以ELISA、RT PCR检测细胞因子表达水平。结果 ELISA结果显示 :IL 2、IL 4、IL 12分泌受到抑制 ,在整个观察期中 ,无论单刺激组还是共刺激组与未接受IL 10干预的对照组比较 ,IL 4、IL 12皆呈现显著差异 ,IL 2分泌在单刺激组抑制显著 ,IFN γ仅表现为明显抑制 ,无统计学差异 ,IL 10促进TGF β1的合成与分泌 ,在单刺激时增加明显 ,而在共刺激组 ,只在反应 72h才显著增加。在基因水平 ,被测细胞因子呈现相似的结果。结论 显性调控的建立主要通过IL 10抑制IL 12和IL 4过度表达 ,防止免疫应答在早期产生特定的和特异的单极化免疫效应。

Objective To observe the dynamic change of cytokines and effect of different stimulating combinations on dominant regulation, and describe the microenviroment character of local immune response. Methods PBMCs were separated from peripheral blood of healthy donors with density gradient centrifugation. The expression levels of cytokine genes under different stimulating combinations, including IL-2, IFN-γ, IL-12, IL-4, IL-10 and TGF-β1, were detected by ELISA and RT-PCR. Results ELISA results indicated that secretion level of IL-2, IL-4, IL-12 and IFN-γ were inhibited by IL-10. There were significant differences in IL-4 and IL-12 between control group and stimulating groups under the stimulation by anti-CD3 monoclonal antibody (McAb) or anti-CD3 McAb + anti-CD28 McAb combination. Significant differences in IL-2 only appeared in anti-CD3 antibody stimulating group. The concentrations of IFN-γ were decreased moderately. At the mean time, IL-10 significantly promoted the secretion of TGF-β1 under the anti-CD3 McAb stimulation. There were similar results on the genes level for the studied cytokines. Conclusion The induction and maintenance of dominant regulation were dependent mainly on IL-10, which inhibits the overexpression of IL-4 and IL-12. It is the vital step for dominant regulation to avoid the mono-polarization development at the beginning of immune response.