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

发布时间：2018-04-25 13:53

本文选题：密码子优化 + 乙型肝炎　；参考：《南京医科大学学报(自然科学版)》2009年03期

【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。
[Abstract]:Aim: to observe whether codon optimization can enhance the immunogenicity of hepatitis B virus surface antigen protein nucleic acid vaccine. Methods: according to the amino acid sequence of the surface antigen of hepatitis B virus (HBV / ADR), a codon optimized gene was designed and synthesized without changing its amino acid sequence. The gene was cloned into nucleic acid vaccine vector pSW3891 and codon optimized MHBs nucleic acid vaccine was constructed. The above nucleic acid vaccine was used to transfect 293T cells with the expressed wild type MHBs nucleic acid vaccine (named as w / pSW3891 / MHBs / adrr) and no-loaded pSW3891. The expression of MHBs in transfected cells was detected by Western blot. BALB/c mice were immunized with optadr and pSW3891 by intramuscular injection. The titer of HBs antibody in serum of immunized mice was detected by ELISA method. The spleen cells secreting IFN- 纬 on the surface antigen of immunized mice were detected by Elispot method. Results both of them were highly expressed in 293T cells. The protein expression of opt was higher than that of wild-type. Opt immunized mice and the number of IFN- 纬 splenocytes secreted by surface antigen of immunized mice was significantly higher than that of adr immunized mice. Conclusion: codon optimization can enhance the protein expression of hepatitis B virus DNA vaccine in vitro and the humoral and cellular immunogenicity of immunized mice.
【作者单位】：南京医科大学第一附属医院感染科;
【基金】：国家自然科学基金资助(30371276)
【分类号】：R392

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