非甲基化CG的寡脱氧核苷酸链对慢性乙型肝炎患者树突状细胞功能的影响

目的:探讨非甲基化CG的寡脱核苷酸链(CPG ODN)对慢性乙型肝炎(CHB)患者树突状细胞(DCs)功能的影响.方法:对10例慢性乙型肝炎患者,10例HBV携带者及10例健康对照者外周血来源DCs进行体外培养.应用流式细胞技术检测DCs的表面分子HLA-DR,CD80和CD86的表达,ELISA检测DCs培养上清中IL-12水平,并在电镜下观察DCs的改变.结果:体外培养第6天的DCs,加入CpG ODN继续培养24 h后,透射电镜下观察,与完全培养基对照组DCs相比,其表面的空起丰富、增粗,粗面内质网增多,空泡减少甚至消失.流式细胞仪分析显示, CpG ODN刺激组CD80,CD86及HLA-DR在CHB患者、HBV携带者及正常人DCs上的表达阳性率均较完全培养基对照组明显升高(P＜0.05).CHB患者、HBV携带者及正常人DCs均用完全培养基培养时,CHB患者及HBV携带者DCs上的HLA-DR,CD86和CD80的表达阳性率较正常人明显下降(P＜0.05),但CHB患者和HBV携带者之间比较差异无统计学意义(P＞0.05).CHB患者、HBV携带者及正常人DCs均用CpG ODN刺激时,CHB患者DCs上的HLA-DR和CD86表达阳性率较HBV携带者及正常人明显下降(P＜0.05),但HBV携带者和正常人之间比较却无统计学差异(P＞0.05);CHB患者和HBV携带者DCs的CD80表达阳性率较正常人明显下降(P＜0.05),但CHB患者和HBV携带者之间比较却无统计学差异(P＞0.05).ELISA检测发现CpG ODN刺激组DCs分泌IL-12的水平无论在CHB患者、HBV携带者及正常人均较完全培养基对照组明显升高(P＜0.05).无论是完全培养基培养还是CpG ODN刺激,CHB患者及HBV携带者DCs分泌IL-12较正常人明显下降(P＜0.05),但CHB患者和HBV携带者之间比较却无统计学差异(P＞0.05).结论:CpG ODN能够明显促进CHB患者外周血DCs成熟,为CpG ODN治疗CHB提供了实验基础.

Effect of CpG motif containing oligodeoxynucleotides on dendritic cell function in patients with chronic hepatitis B

Objective To explore the effect of unmethylated CpG motif containing oligodeoxynucleotides (CpG ODN) on the function of dendritic cells (DCs) in patients with chronic hepatitis B (CHB). Methods DCs were obtained from peripheral blood mononuclear cells (PBMCs) of 15 CHB patients, 12 hepatitis B virus (HBV) carriers, and 10 healthy controls. The expressions of HLA-DR, CD80, and CD86 on DCs were determined by fluorescence activated cell sorting (FACS). The IL-12 level in supernatant of the culture medium was measured by ELISA, and the morphological changes of DCs were observed under transmission electron microscope. Results Compared with the controls, DCs stimulated with CpG ODN represented enrichment in cell surface protrusions and rough endoplasmic reticulum, decreased or disappeared vacuole. The expressions of HLA-DR, CD86, and CD80 were much higher in DCs stimulated with CpG ODN than those in complete medium control (P＜0.05). When culturing in complete medium, the expressions of HLA-DR, CD86, and CD80 were much lower in CHB patients and HBV carriers than healthy controls (P＜0.05). The expressions of HLA-DR and CD86 stimulated with CpG ODN were much lower in CHB patients than HBV carriers and healthy controls (P＜0.05). The expressions of CD80 were much lower in CHB patients and HBV carriers than healthy controls (P＜0.05). The levels of IL-12 were much higher in DCs stimulated with CpG ODN than that in complete medium controls (P＜0.05). The levels of IL-12 in complete medium or medium added with CpG ODN were much lower in CHB patients and HBV carriers than in healthy controls (P＜0.05).Conclusion CpG ODN could significantly promote the maturation of dendritic cells in peripheral blood in CHB patients.