慢性乙型肝炎患者树突状细胞体外功能恢复及诱导T细胞反应研究

Rescue of dendritic cells malfunction and loading with HBV proteins in patients with chronic hepatitis B in vitro

目的研究慢性乙型肝炎（CHB）患者外周血树突状细胞（DC）和T细胞体外功能，并尝试恢复功能的方法。方法12例CHB患者作为研究对象，10例健康人作为正常对照研究。对外周血DC进行表型分析和混合淋巴细胞反应（MLR）测定，并分别用HBsAg或HBcAg作为抗原负载DC，后用ELIspot法检测产生IFN-7的细胞频数。结果CHB患者外周血DC共刺激分子水平表达降低，刺激MLR能力不足。经过体外补充促成熟细胞因子，DC缺陷能够得到部分纠正。成熟DC较不成熟DC刺激MLR和诱导产生IFN-γ的T细胞能力强，HBcAg比HBsAg有更强的免疫原性。结论慢性CHB患者外周血DC功能缺陷，能够通过补充促成熟因子纠正。成熟DC比不成熟DC显示出更强的MLR反应和诱导IFN-γ产生细胞的能力，这提示HBV蛋白负载DC作为治疗性疫苗具有可行性。

Objective To investigate dendritic cell （DC） malfunctions in patients with chronic hepatitis B （CHB） and try some means to restore the function in vitro. Methods Twelve CHB patients and 10 healthy people were enrolled in the study. Phenotype analysis and allogeneic mixed lymphocyte reaction （AMLR） assay of DC from these subjects were made. Enzyme-linked ELISpot method for detecting IFN-γ-producing CD8 （ ＋ ） T cells were used to evaluate the efficacy of DC loaded in vitro with HBsAg or HBcAg. Results DC from patients had a lower expression of co-stimulatory molecules and impaired AMLR capacity, but was restored partially by cytokine cocktail in vitro. Mature DC loaded with HBsAg or HBcAg showed a greater capacity for IFN-γ-production than immature DC. Conclusion Malfunction of DC from CHB patients may be rescued by a cocktail of cytokines, and therapeutic DC vaccines loaded with HBV protein might be helpful to treat CHB natients.