CpG寡核苷酸联合HBsAg对慢性乙型肝炎患者树突细胞功能及核因子活性的影响

Effects of CpG-ODN combined with HBsAg on the phenotype, function and the activity of NF-kappa B and AP-1 of monocyte-derived dendritic cells in chronic hepatitis B patients

目的研究含CpG寡核苷酸（CpG—ODN）联合重组HBsAg对慢性乙型肝炎（CHB）患者外周血树突状细胞（DC）表型和功能及其对核因子-κB和激活蛋白-1活性的影响。方法以重组细胞因子联合诱导扩增CHB患者外周血单个核细胞得到DC；以CpG—ODN和HBsAg及肿瘤坏死因子α刺激DC，评价其表型、功能和胞核核因子-κB、激活蛋白-1活性。结果CpG—ODN联合HBsAg明显提高CHB患者DC表面分子人类白细胞抗原-DR表达、白细胞介素-12分泌以及刺激同种T淋巴细胞增殖的能力，尤其能提高CD1a的表达。上述免疫佐剂能增强DC细胞核因子-κB活性，同时抑制激活蛋白-1活性。结论CpG—ODN与hTNF α一样能够促进CHB患者外周血DC成熟；其作用机制可能是选择性激活或抑制DC胞内不同核因子活性。

Objective To investigate the effect of synthetic non-methylated CpG-ODN combined with recombined HBsAg on the phenotype, function and the expression level of nucleic transcription factor NF- kappa B and AP-1 of monocyte-derived dendritic cells （DC） in patients with chronic hepatitis B （CHB）. Methods Purified monocytes were isolated from the peripheral blood of CHB patients and healthy volunteers and cultured with human granulocyte-monocyte colony stimulating factor added together with intedeukin- 4. On the fifth day of culture, CpG-ODN, HBsAg and other reagents, such as TNF alpha and PBS, were added to the medium, and 18 hours later cells were collected for the detection of surface molecules （HLA-DR/ CD86/CD1a）. IL-12p70 levels in the supernatant and stimulating capacity to allogenic T lymphocytes were detected. The nucleic proteins of NF-kappa B and AP-1 in DC were extracted and purified for the gel shift assay. Results Compared with those of the PBS group, the expression rates of HLA-DR of DC treated with CpG-ODN and/or HBsAg were obviously increased, Both the IL-12p70 level and the stimulating capacity of DC to allogenic T lymphocytes in mixed lymphocyte reaction increased significantly in the CpG-ODN group and in the CpG-ODN/HBsAg combination group （P 〈 0,01 and 0.05, respectively）, The expression rates of CD1a were raised only in the CpG-ODN/HBsAg combination group, Three kinds of immunological adjuvants, TNF alpha, CpG-ODN and CpG-ODN/HBsAg enhanced the expression of nucleic NF-kappa B and inhibited the expression of AP-1 in DC. Conelusion CpG-ODN, like TNF alpha, has remarkable stimulatory effect on the impaired phenotype and function of monocyte-derived DC in patients with CHB; CpG-ODN and HBsAg have a synergetic effect in increasing the antigen presenting function, The regulating ability of CpG-ODN and TNF alpha on the expression levels of NF-kappa B and AP-1 might be one of the mechanisms of their immunostimulatory effects on DC of the CHB patients.