霉酚酸衍生物对人PBMC产生IL-17和IFN-γ的抑制作用

The inhibition effect of mycophenolic acid derivative on production of IL17 and IFN-γ by human peripheral blood mononuclear cell

目的探讨霉酚酸衍生物(JU95-07B)对人外周血单个核细胞(PBMC)产生IL-17和IFN-γ的抑制作用,为临床治疗炎症及自身免疫性疾病提供依据。方法用anti-CD3+anti-CD28刺激PBMC,酶联免疫吸附法(ELISA)检测加入不同浓度JU95-07B及作用不同时间点的IL-17和IFN-γ水平。用流式细胞术(FCM)检测JU95-07B对T细胞不同亚群表达IL-17和IFN-γ的影响。用RT-PCR法测定JU95-07B对IL-17和IFN-γmRNA的作用。结果 JU95-07B对anti-CD3+anti-CD28诱导PBMC产生的IL-17和IFN-γ均呈剂量依赖性的抑制,与作用时间有关,早期无明显的抑制现象,12 h后开始出现有统计学意义的抑制效果(P<0.05),培养时间越长抑制效果越明显。JU95-07B可明显降低CD4+和CD8+T细胞产生IL-17和IFN-γ的比例,对IL-17的抑制率高于IFN-γ。此外,JU95-07B抑制IL-17和IFN-γmRNA的表达。结论 JU95-07B能有效抑制T细胞产生IL-17和IFN-γ。

Objective To study the effect of mycophenolic acid derivative （JU95-O7B） on the production of IL-17 and IFN-y by human PBMC. Methods PBMC were cultured with anti-CD3 plus anti-CD28 in the absence or presence of JU95-07B at different concentrations or different time points. of IL-17 and IFN-y by enzyme-linked immunosorbent assay The supernatants were harvested and detected for production （ELISA）. Flow cytometry （FCM） was used to detect the effect of JU95-07B on the expression of IL-17 and IFN-yfrom different T cells subsets. Furthermore,the mRNA levels of IL-17 and IFN-y were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results After stimulation with anti-CD3 plus anti-CD28, PBMC produced high levels of IL-17 and IFN-y, but was inhibited by JU95-07B in a dose dependent manner, and also associated with the length of cultures. There were no significant inhibitory effects on the production of cytokines within 8 h. However, the inhibition of IL-17 and IFN-y,/ production in PBMC by JU95-O7B occurred at 12 h, the longer the culture, the stronger the inhibition. JU95-07B could significantly reduce the ratio of IL- l7 and IFN-y-producing CD4＋ and CD8＋ T cell. In addition, JU95-07B also inhibited the expression of IL-17 and IFN- mRNA. Conclusion JU95-07B can effectively inhibit the production of IL-17 and IFN-yfrom T cells.