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目的 为激发机体对ＨＢｓＡｇ的ＣＴＬ反应，寻求对慢性乙型肝炎更有效的治疗方法。方法 构建了共表达ＨＢＳＡｇ及Ｂ７－２抗原的Ｅ１区插入重组腺病毒载体，用脂质体法转染２９３细胞，经空斑筛选表达目的抗原的重组腺病毒，用ＥＬＩＳＡ法及Ｗｅｓｔｅｒｎ ｂｌｏｔｔｉｎｇ法分别检测ＨＢｓＡｇ及Ｂ７－２抗原表达。在２９３细胞中扩增，再纯化、浓缩后，体内感染Ｃ５７小鼠，检测体液免疫反应和细胞免疫反应。结果 重组腺病毒在体外培养的２９３细胞及ＨｅｐＧ２细胞中高效表达ＨＢＳＡｇ及Ｂ７－２抗原。感染小鼠体液免疫较弱，但可通过注射乙型肝炎疫苗而加强；重组腺病毒载体能诱导强而有效的细胞免疫反应；未观察到明显毒副作用。结论 ＨＢｓＡｇ及Ｂ７－２抗原重组腺病毒载体体外感染２９３、ＨｅｐＧ２细胞后高效表达目的抗原，感染免疫局诱导有效的细胞免疫反应。初步结果显示腺病毒载体是一种高效、安全的载体系统。
The purpose is to stimulate the body’s CTL response to HBsAg and to seek a more effective treatment for chronic hepatitis B. Methods Recombinant adenoviral vector was constructed by inserting the E1 region coexpressing HBsAg and B7-2 antigen into 293 cells. The recombinant adenovirus expressing the antigen of interest was screened by plaque. ELISA and Western blotting were performed respectively Detection of HBsAg and B7-2 antigen expression. After amplification in 293 cells, re-purification and concentration, C57 mice were infected in vivo to detect humoral and cellular immune responses. Results Recombinant adenovirus highly expressed HBSAg and B7-2 antigen in 293 cells and HepG2 cells cultured in vitro. Infected mice had weaker humoral immunity, but could be strengthened by injecting Hepatitis B vaccine. Recombinant adenovirus vector could induce strong and effective cellular immune response; no obvious side effects were observed. Conclusions The recombinant adenovirus vector of HBsAg and B7-2 antigen infect 293 and HepG2 cells in vitro and express the antigen of interest efficiently. Infection immunity Bureau induces effective cellular immune response. Preliminary results show that adenovirus vector is an efficient and safe vector system.