论文部分内容阅读
目的探讨人白细胞介素 15 (humaninterleukin 15 ,hIL 15 )基因佐剂对柯萨奇B3病毒 (CoxsackievirusB3 ,CVB3 )VP1基因疫苗的免疫增强效果。方法从人胚肺二倍体细胞提取hIL 15的mRNA ,用逆转录 聚合酶链反应 (reversetranscriptase polymerasechainreaction ,RT PCR)扩增出cDNA ,构建成真核表达克隆 pcDNA3 IL 15。将pcDNA3 VP1、pcDNA3 VP1+ pcDNA3 IL 15、pcDNA3 IL 15、pcDNA3和盐水分别肌内注射Balb/c小鼠 ,每次免疫后 6天取血清用微量中和试验测CVB3中和抗体 ,3次免疫后用 80 0TCID5 0CVB3感染小鼠 ,观察小鼠的生存率。结果 pcDNA3 IL 15重组质粒的目的基因与hIL 15基因序列相同 ;pcDNA3 VP1和pcDNA3 VP1+ pcDNA3 IL 15能诱导小鼠产生中和抗体 ,抗体滴度随免疫次数增加而提高 ;pcDNA3 VP1+ pcDNA3 IL 15组抗体滴度和生存率均高于 pcDNA3 VP1组。结论本研究成功构建了hIL 15重组质粒 ,该质粒对真核表达重组质粒 pcDNA3 VP1有一定的免疫原性增强作用
Objective To investigate the effect of human interleukin 15 (hIL 15) adjuvant on VP1 gene vaccine of Coxsackievirus B3 (CVB3). Methods mRNA of hIL 15 was extracted from human embryo lung diploid cells. The cDNA of hIL 15 was amplified by reverse transcriptase polymerase chain reaction (RT PCR) and cloned into eukaryotic expression plasmid pcDNA3 IL 15. Balb / c mice were intramuscularly injected with pcDNA3 VP1, pcDNA3 VP1 + pcDNA3 IL15, pcDNA3 IL15, pcDNA3 and saline respectively. After 6 days of each immunization, the serum was neutralized with CVB3 by neutralization test. After three immunizations Mice were infected with 80 0 TCID5 0CVB3 to observe the survival rate of mice. Results The target gene of pcDNA3 IL 15 recombinant plasmid was the same as that of hIL 15 gene. The pcDNA3 VP1 and pcDNA3 VP1 + pcDNA3 IL 15 could induce neutralizing antibodies in mice. The antibody titers increased with the increase of the number of immunizations. The antibody against pcDNA3 VP1 + pcDNA3 IL 15 Titers and survival rates were higher than pcDNA3 VP1 group. Conclusion The hIL 15 recombinant plasmid was successfully constructed in this study. The recombinant plasmid has eukaryotic expression of recombinant plasmid pcDNA3 VP1 to enhance immunogenicity