文章摘要
杜合娟,王世霞,李 军,黄祖瑚,卢 山,邢益平.密码子优化对乙肝病毒表面抗原中蛋白核酸疫苗免疫原性的影响[J].南京医科大学学报,2009,29(3):286~290
密码子优化对乙肝病毒表面抗原中蛋白核酸疫苗免疫原性的影响
The effect of codon optimization on the immunogenicity of middle hepatitis B surface antigen DNA vaccine
投稿时间:2008-09-23  
DOI:10.7655
中文关键词: 密码子优化  乙型肝炎  表面抗原  核酸疫苗  免疫原性
英文关键词: codon optimization  hepatitis B  surface antigen  DNA vaccine immunogenicity
基金项目:国家自然科学基金资助(30371276)
作者单位
杜合娟 南京医科大学第一附属医院感染科,江苏 南京 210029 
王世霞  
李 军  
黄祖瑚  
卢 山  
邢益平  
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中文摘要:
      目的:观察密码子优化能否增强乙型肝炎病毒表面抗原中蛋白核酸疫苗的免疫原性?方法:根据乙型肝炎病毒(adr亚型)表面抗原中蛋白(MHBs)的氨基酸序列,在不改变其氨基酸序列的基础上,设计并人工合成了密码子优化的基因,并将该基因克隆到核酸疫苗载体pSW3891中,构建了密码子优化的表达MHBs 核酸疫苗(命名为:pSW3891/MHBs/adr/opt,简称opt)?用上述核酸疫苗与表达野生型MHBs核酸疫苗(命名为:pSW3891/MHBs/adr,简称adr)及空载体质粒pSW3891一起瞬时转染293T 细胞,应用蛋白质印迹法检测转染细胞中MHBs的表达?采用肌肉注射法,以opt?adr及pSW3891,分别对BALB/c小鼠进行免疫?用ELISA方法检测免疫后小鼠血清中HBs抗体的滴度,ELISPOT法检测免疫小鼠表面抗原特异性分泌 IFN-γ的脾细胞?结果:opt和adr均可在体外293T 细胞中高效表达,且opt的蛋白表达量要高于野生型?opt免疫组小鼠特异性抗体滴度和免疫小鼠表面抗原特异性分泌 IFN-γ脾细胞数量都要显著的高于adr免疫组小鼠?结论:密码子优化可以增强乙型肝炎病毒DNA疫苗在体外的蛋白表达量及免疫小鼠的体液免疫和细胞免疫原性?
英文摘要:
      Objective:To investegate whether codon optimization of middle hepatitis B surface antigen could enhance the immunogenicity of HBV DNA vaccine. Methods:According to the sequence of amino acids of MHBs(adr subtype), the codon optimized genes were designed and synthesized without changing the sequence of amino acids and then cloned into vector pSW3891, which was named pSW3891/MHBs/adr/opt. 293T cells were transiently transfected with opt and wild-type MHBs DNA vaccine(pSW3891/MHBs/adr) and empty vector pSW3891. The protein expression level was assessed by western blot. BALB/c mice were intramuscularly injected with opt, adr or pSW3891. Anti-HBs in sera was tested by ELISA. IFN-γsecretion splenocytes of mice immunized with opt, adr and pSW3891 were tested by ELISPOT. Results:The level of protein expression in both supernatant and lysate of 293T cells transfected with opt is higher than that in 293T cells transfected with adr. The BALB/c mice immunized with opt showed stronger anti-HBs response and more IFN-γsecretion splenocytes than those immunized with adr. Conclusion:Codon optimization of MHBs DNA vaccine could improve protein expression and induce significant humoral and cellular immune responses in BALB/c mice.
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