文章摘要
张 璐,张春华,李 军,黄祖瑚,卢 山,王世霞.密码子优化的中国HIV-1流行亚型gp120共识序列核酸疫苗的构建及其免疫原性的初步研究[J].南京医科大学学报,2008,28(9):1104~1108
密码子优化的中国HIV-1流行亚型gp120共识序列核酸疫苗的构建及其免疫原性的初步研究
Construction and immunogenicity of codon optimized DNA vaccines expressing consensus gp120 of HIV-1 subtypes in China
  
DOI:10.7655
中文关键词: 疫苗  核酸疫苗  HIV-1  gp120  密码子优化
英文关键词: vaccine  DNA vaccine  HIV-1  gp120  codon optimization
基金项目:江苏省感染病学重点实验室(传染病临床医学中心)资助[苏卫规财(2007)158]
作者单位
张 璐 南京医科大学第一附属医院中美疫苗研究中心,江苏省感染病学重点实验室,江苏 南京 210029 
张春华 南京医科大学第一附属医院中美疫苗研究中心,江苏省感染病学重点实验室,江苏 南京 210029 
李 军 南京医科大学第一附属医院中美疫苗研究中心,江苏省感染病学重点实验室,江苏 南京 210029 
黄祖瑚 南京医科大学第一附属医院中美疫苗研究中心,江苏省感染病学重点实验室,江苏 南京 210029 
卢 山 美国马萨诸塞州大学医学院内科核酸疫苗研究室,美国 渥斯特 01605 
王世霞 美国马萨诸塞州大学医学院内科核酸疫苗研究室,美国 渥斯特 01605 
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中文摘要:
      目的:构建能表达密码子优化的中国HIV-1流行亚型AE?BC和B’gp120共识序列的核酸疫苗,并研究其免疫原性?方法:根据HIV-1的分子流行病学资料和包膜糖蛋白的氨基酸序列分析,设计了针对中国流行的AE,BC和B’(ThB)亚型的gp120共识序列?按哺乳动物细胞偏好的密码子对AE,BC和ThB 3个亚型的gp120共识序列分别进行密码子优化,合成优化基因?将密码子优化的gp120基因克隆到核酸疫苗载体pSW3891,构建重组质粒核酸疫苗?将重组质粒转染293T 细胞,并以重组质粒疫苗免疫新西兰兔?应用蛋白质免疫印迹法检测转染细胞中gp120蛋白的表达?ELISA方法检测免疫后兔血清中gp120特异性抗体的产生?结果:3个密码子优化的gp120核酸疫苗,均能正确表达gp120?这些gp120核酸疫苗免疫家兔后,能产生HIV-1包膜糖蛋白特异性的抗体?结论:成功构建3个密码子优化的?表达HIV-1中国流行亚型AE,BC和ThB 的共识序列的gp120核酸疫苗,能诱导HIV-1 包膜蛋白特异的免疫反应?
英文摘要:
      Objective:To construct and test the immunogenicity of codon-optimized DNA vaccines expressing gp120 from key representative HIV-1 subtypes AE,BC and B′(ThB) in China. Methods:Based on HIV-1 epidemiological information and sequence analysis of HIV-1 envelope glycoprotein(Env) of subtypes AE,BC and ThB in China,we designed the consensus gp120 sequence for each subtype AE,BC or ThB. These consensus gp120 genes were codon-optimized using codons preferred for high level expression in mammalian systems and chemically synthesized. The codon optimized gp120 genes were subcloned into the DNA vaccine vector pSW3891. The expression of gp120 by the constructed DNA vaccine plasmids in 293T cells was confirmed by Western blot. New Zealand rabbits were immunized with gp120 DNA vaccines using electroporation method,and gp120-specific antibodies in rabbit sera were evaluated by ELISA. Results:In the codon optimized gp120 gene,the frequency of the mammalian cell preferred codons was higher than that of the wild type gp120 gene. Three codon optimized gp120 DNA vaccines of consensus subtypes AE,BC or ThB were constructed and the expression of gp120 antigens was confirmed in transiently transfected 293T cells. The rabbits immunized with these gp120 DNA vaccines elicited HIV-1 Env-specific antibody responses. Conclusion:Three codon optimized DNA vaccines expressing consensus gp120 of HIV-1 sybtypes AE,BC and ThB were successfully constructed with proven expression and immunogenicity in eliciting HIV-1 Env-specific humoral responses.
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