HCV核心-包膜E2抗原融合基因DNA疫苗的构建及对小鼠的免疫应答试验  

Construction of HCV core and envelope 2 fusion gene DNA vaccine and its immunogenicity in mice

作　　者：姜春鹏[1] 赵平[2] 朱诗应[2] 王路[2] 戚中田[2] 

机构地区：[1]东北农业大学生物工程系 [2]第二军医大学微生物学教研室

出　　处：《中华微生物学和免疫学杂志》2002年第5期510-514,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 (3 9980 0 3 8);军队"十五"医药卫生科研基金资助项目 (0 1MA160 )

摘　　要：目的　观察一种结构新颖的HCV融合抗原DNA疫苗在BALB c小鼠的免疫效果 ,探讨其用于防治丙型肝炎的可行性。方法　用重叠延伸PCR拼接编码小鼠IgGkappa链信号肽和通用型辅助性T细胞表位PADRE的DNA片段 ,PCR分别扩增HCV核心抗原基因和包膜E2抗原基因 ,将 3段基因插入真核表达载体pcDNA3.1,构成重组表达质粒pST CE2t,转染COS7细胞 ,免疫组化检测HCV抗原的表达。将pST CE2t和HCV核心抗原DNA疫苗pcDNA3.1core分别肌肉注射接种BALB c小鼠 ,检测小鼠的血清抗体、T细胞增殖和CTL反应。结果　pST CE2t可在COS7细胞内表达HCV核心抗原和E2抗原 ,接种于BALB c小鼠能有效诱导体液和细胞免疫应答 ,其中抗HCV核心抗原免疫应答的强度明显超过pcDNA3.1core ,且更趋向于TH1型免疫应答。结论　pSTObjective To observe the immunogenicity of a novel hepatitis C virus (HCV)fusion protein DNA vaccine in BALB/c mice and determine its value as an agent against hepatitis C. Methods A DNA fragment encoding signal peptide of murine IgG kappa chain and a universal helper T lymphocyte epitope PADRE were spliced by overlapping PCR. HCV core gene and envelope 2 (E2)gene were amplified by PCR respectively. The three DNA fragments were then inserted into an eukaryotic expression vector pcDNA3.1. The resultant recombinant expression plasmid pST CE2t was transfected into COS7 cells and its expressed product was identified by immunohistochemical staining. BALB/c mice were intramuscularly vaccinated with pST CE2t or pcDNA3.1 core (HCV core DNA vaccine) three times. The serum antibodies, T lymphocyte proliferative response and cytotoxic T lymphocyte response of the mice were examined. Results pST CE2t expressed HCV core and E2 antigens in COS7 cells and induced humoral as well as cellular immune responses in BALB/c mice effectively. The immune responses against HCV core protein elicited by pST CE2t are stronger and more inclined to T H1 type than that of pcDNA3.1 core. Conclusion This optimal HCV core and E2 fusion protein DNA vaccine may be potentially used for prevention and treatment of hepatitis C. [

关 键 词：HCV 丙型肝炎病毒 丙型肝炎 DNA疫苗 核心抗原 包膜E2抗原 免疫应答 

分 类 号：R392[医药卫生—免疫学]