乙型肝炎病毒治疗性多表位基因疫苗在小鼠中的免疫应答研究  

Study on the immune response of the therapeutic multi-epitope gene vaccine of hepatitis B virus in mice

作　　者：骆利敏 夏虎[2] 刘树人 李明[3] 

机构地区：[1]解放军458医院肝病中心传染科 [2]南方医科大学附属珠江医院呼吸科 [3]南方医科大学生物技术学院,广东广州510602

出　　处：《细胞与分子免疫学杂志》2011年第3期260-262,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30170532);广东省自然科学基金资助项目(990375);广州市重点攻关课题资助项目(2001-2-035-01)

摘　　要：目的:分析乙型肝炎病毒(HBV)重组多表位抗原基因在大肠杆菌中非融合表达的蛋白B-BPT的免疫原性,以期为HBV预防/治疗性疫苗的研制奠定实验基础。方法:构建重组质粒pBAD/BPT,并在大肠杆菌中诱导表达。以纯化的蛋白B-BPT免疫BALB/c小鼠,用乳酸脱氢酶(LDH)释放法检测该融合蛋白诱导的小鼠细胞毒T淋巴细胞(CTL)应答。用流式细胞术(FCM)检测小鼠CD4+、CD8+T淋巴细胞亚群的比例,用MTT比色法检测淋巴细胞的增殖效应。结果:B-BPT蛋白免疫BALB/c小鼠后,可诱导特异性CTL应答(P<0.05,与空白对照组比较);B-BPT蛋白免疫后,可显著刺激小鼠脾淋巴细胞增值(P<0.05,与空白对照组比较)。结论:该非融合表达的蛋白B-BPT可有效地诱导小鼠特异性CTL免疫应答,为进一步研制乙肝预防治疗性疫苗奠定了基础。AIM： To study the immunogenicity of the therapeutic multi-epitope gene vaccine of Hepatitis B virus.METHODS： Multi-epitope gene of Hepatitis B virus was cloned into prokaryon expression vector pBAD/gIIIA to express a non-fusional antigen B-BPT and BALB/c mice were immunized with the antigen.Cytotoxic T-lymphocyte（CTL） was determined with CytoTox96 kit which quantitatively measures the lactate dehydrogenase（LDH） that is released on cell lysis.CD4＋,CD8＋ T lymphocytes subsets in immunized mice was detected by using flow cytometry（FCM）.Lymphocyte proliferation response was completed by MTT colorimetry.RESULTS： Injection of B-BPT elicited high-level of CTL response and also stimulus spleen lymphocytes to proliferate effectively.CONCLUSION： B-BPT can induce specific cellular immune responses and may be a good candidate for therapeutic vaccine.

关 键 词：乙型肝炎病毒 治疗性疫苗 多表位抗原 

分 类 号：R979.5[医药卫生—药品]