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目的探讨体外HBeAg特异性细胞免疫反应对乙型肝炎病毒(HBV)mRNA表达及蛋白合成的影响。方法自正常人外周血单核细胞中诱导培养树突状细胞(DC),经不同浓度的HBeAg负载后与自身T细胞共同培养,诱导产生HBeAg特异性T细胞,同时设与无HBeAg刺激的DC共同培养的T细胞、单纯T细胞为对照组,作用于HepG2.2.15细胞系,逆转录PCR(RT-PCR)检测HepG2.2.15细胞内乙肝表面抗原(HBsAg)、乙肝核心抗原(HBcAg)的mRNA水平,酶联免疫法(ELISA)检测细胞培养上清中HB-sAg和乙肝e抗原(HBeAg)水平。结果经HBeAg刺激激活的T细胞可以有效地抑制HBsAg、HBcAg mRNA的表达(P<0.05)及HBsAg、HBeAg的合成(P<0.05),且随着HBeAg刺激浓度的升高,对上述指标的抑制率逐渐上升。结论HBeAg特异性细胞免疫反应可显著抑制HBV mRNA和蛋白的表达,为开发HBeAg相关的治疗性疫苗奠定理论基础。
Objective To investigate the effect of HBeAg-specific cellular immune response on the expression of hepatitis B virus (HBV) mRNA and protein synthesis in vitro. Methods Dendritic cells (DCs) were induced from normal human peripheral blood mononuclear cells (DCs). After different concentrations of HBeAg were loaded and co-cultured with their own T cells, HBeAg-specific T cells were induced. Meanwhile, DCs stimulated with HBeAg Co-cultured T cells and pure T cells were used as controls in HepG2.2.15 cell line. The mRNA expression of HBsAg and HBcAg in HepG2.2.15 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) The levels of HBsAg and HBeAg in the cell culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA). Results The T cells stimulated by HBeAg could effectively inhibit the expression of HBsAg and HBcAg (P <0.05) and the synthesis of HBsAg and HBeAg (P <0.05), and inhibit the above mentioned indexes with the increase of HBeAg stimulation The rate gradually increased. Conclusion HBeAg-specific cellular immune response can significantly inhibit the expression of HBV mRNA and protein, which lays a theoretical foundation for the development of HBeAg-related therapeutic vaccines.