摘要
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8 d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IFNγ、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IFNγ、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。
Objective To study the maturation of dendritic cells and the activity of CD4＋/CD8＋T cells after prostaglandin E1 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 E1 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,IL-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 E1 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 E1 control group,prostaglandin E1could down-regulate the expression of the surface markers on dendritic cells,including HLA-DR,CD83 and CD86,and inhibit the secretion of IL-12p70.In addition,prostaglandin E1 could inhibit the cytotoxic activity of the T lymphocyte in vitro.Conclusion Prostaglandin E1 inhibited the activation of CD4＋/CD8＋T cells,the maturation of dendritic cells and the cytotoxic activity of T lymphocytes.
出处
《临床肝胆病杂志》
CAS
2012年第3期212-215,共4页
Journal of Clinical Hepatology
关键词
前列地尔
肝炎
乙型
慢性
树突细胞
alprostadil
hepatitis B
chronic
dendritic cells
