Xing Li-he, Wang Fu-sheng, Zhu Chuan-lin
Institute of Infectious Diseases, The No.302 Hospital of PLA, Beijing, 100039, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2003 Dec;17(4):365-8.
To investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant.
DCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method.
DCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg.
Human dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.
研究人树突状细胞（DCs）诱导特异性T淋巴细胞反应的能力以及抑制2.2.15细胞培养上清液中HBeAg和HBsAg表达的能力。
采用粒细胞巨噬细胞集落刺激因子（GM-CSF）和白细胞介素4诱导外周血单个核细胞制备DCs。在DCs成熟前用纯HBsAg冲击DCs，并与自身血T淋巴细胞共培养，同时制备未用纯HBsAg刺激的DCs组、T淋巴细胞组和单纯T淋巴细胞组作为对照组。分别在第1天、第3天、第5天和第7天收集经刺激的T淋巴细胞培养的2.2.15细胞的培养上清液。采用ELISA法检测HBeAg和HBsAg的表达水平。
抗原刺激后的DCs具有较强的抗原呈递能力和诱导免疫激活能力，正常对照组和慢性肝炎患者组中加载抗原后的DCs对T淋巴细胞增殖的刺激能力强于未加载抗原的DCs组和单纯T淋巴细胞组（P<0.01）。DCs的刺激能力与加载抗原的剂量呈正相关；DCs刺激产生的CTLs对2.2.15细胞上清液中HBeAg的表达有特异性抑制作用，但对HbsAg的表达无抑制作用。
体外HBsAg刺激的人树突状细胞能有效呈递抗原并刺激产生针对HBV的特异性CTLs，可有效抑制2.2.15细胞上清液中HBeAg的表达——DC疫苗未来可能成为慢性乙型肝炎病毒感染患者的抗病毒治疗策略。
