前列腺素E1抑制慢性乙型重型肝炎树突状细胞成熟及CD4+/CD8+T细胞活性的观察

摘要

Objective To study the maturation of dendritic cells and the activity of CD4 + /CD8 + T cells after prostagl andin El treatment in chronic severe hepatitis B patients. Methods PBMC were isolated from fresh peripheral blood of chronic severe hepatitis B patients by Ficoll - Hypaque density gradient centrifugation and cultured with plastic - adherence method. Granulocyte - macrophage colony stimulating factor and interleukin -4 with or without prostaglandin El were used to induce the PBMC to obtain dendritic cells for 8 days. FACS was used to analyze the expression of the DC surface markers, including human leukocyte antigen ( HLA - DR) , CD83, CD86. The secretion of IFN - γ, JL - 12p70 was detected by ELISA. Some of the non -adherent cells were used to analyze the ratio of CD4+/CD8+ T cells and the expression of CD69 and HLA -DR, the others were incubated with IL-2 for analyzing the cytotoxic T lymphocyte activity. Results Although prostaglandin El had no effects on the activity of CD4+/CD8+T cells in chronic severe hepatitis B patients, it could decrease the expression of CD4+ /CD8+ T cells activators, such as CD69 and HLA - DR. Further more, compared with the non - prostaglandin El control group, prostaglandin El could down - regulate the expression of the surface markers on dendritic cells, including HLA -DR, CD83 and CD86, and inhibit ihe secretion of IL - 12p75. In addition, prostaglandin El could inhibit the cytotoxie activity of the T lymphocyte in vitro. Conclusion Prostaglandin El inhibited the activation of CD4+ /CD8+ T cells, the maturation of dendritic cells and the cytotoxie activity of T lymphocytes.%目的 研究前列腺素E1对慢性乙型重型肝炎树突状细胞成熟及CD4 +/CD8+T细胞活性的影响.方法 抽取前列腺素E1治疗10 d前后的慢性乙型重型肝炎患者外周静脉血,密度梯度离心法分离淋巴细胞,贴壁培养获得PBMC,经重组人白细胞介素4(rhIL-4)、重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)、加或不加前列腺素E1刺激培养8d获得树突细胞(DC).经流式细胞仪检测DC表达的HLA-DR、CD83、CD86,ELISA测定分泌的IFN γ、IL-12p70.悬浮T细胞部分检测CD4 +/CD8+T细胞比例及其细胞表面CD69、HLA-DR表达；部分经IL-2培养,用于检测DC体外诱导的T淋巴细胞毒活性.结果前列腺素E1治疗慢性乙型重型肝炎,对患者CD4 +/CD8+T细胞比例无明显影响,可抑制CD4 +/CD8+T细胞活化分子CD69、HLA-DR的表达；前列腺素E1体外培养的DC低表达HLA-DR、CD83、CD86,分泌IFN γ、IL-12p70均下降,体外诱导的自体淋巴细胞毒活性减弱,与未加用前列腺素E1比较差异有统计学意义(P＜0.01).结论 前列腺素E1治疗慢性乙型重型肝炎,可抑制CD4 +/CD8+T细胞活化；体外培养可抑制DC成熟,诱导减弱自体淋巴细胞毒活性.