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目的探讨人乳头瘤病毒58型(HPV58)早期基因E2作为DNA疫苗候选抗原的可行性。方法构建HPV58E2真核重组表达载体pcDNA3/58E2,转染SiHa细胞,利用RT-PCR检测HPV58E2mRNA的转录。pcDNA3/58E2DNA肌肉注射免疫BALB/C小鼠,ELISA检测小鼠血清中特异性anti-HPV58E2IgG水平。结果RT-PCR结果表明,pcD-NA3/58E2能够在真核细胞中有效转录HPV58E2mRNA。动物实验表明,质粒pcDNA3/58E2肌肉接种可诱导免疫小鼠产生抗E2特异性抗体。结论本研究成功构建了重组真核表达载体pcDNA3/58E2,并初步证明HPV58E2DNA疫苗有良好的免疫原性。
Objective To investigate the feasibility of using human papillomavirus type 58 (HPV58) early gene E2 as a DNA vaccine candidate antigen. Methods The recombinant eukaryotic expression vector pcDNA3 / 58E2 was constructed and transfected into SiHa cells. The transcription of HPV58E2 mRNA was detected by RT-PCR. BALB / C mice were immunized with pcDNA3 / 58E2 DNA by intramuscular injection. The level of specific anti-HPV58E2 IgG in serum was detected by ELISA. Results RT-PCR results showed that pcD-NA3 / 58E2 efficiently transcribed HPV58E2 mRNA in eukaryotic cells. Animal experiments show that the plasmid pcDNA3 / 58E2 muscle inoculation can induce immune mice produce anti-E2 specific antibodies. Conclusion This study successfully constructed recombinant eukaryotic expression vector pcDNA3 / 58E2, and preliminary evidence of HPV58E2 DNA vaccine has good immunogenicity.