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【目的】以减毒鼠伤寒沙门菌(Salmonella typhimurium aroA strain SL7207,SL7207)为载体携带可表达呼吸道合胞病毒(Human respiratory syncytial vrius,RSV)密码子优化的融合蛋白(Fusion glycoprotein,F)的真核表达质粒,探讨不同黏膜免疫途径及密码子优化对免疫效果的影响。【方法】通过对RSV野生型F基因(Fwt)进行密码子优化,获得密码子优化的F基因(Fsyn),并构建可表达Fsyn的真核表达质粒pcDNA3.1/Fsyn,转化SL7207得到SL7207/pcDNA3.1/Fsyn。分别经滴鼻和灌胃途径,免疫BALB/c小鼠,采用间接ELISA方法比较免疫效果。【结果】与灌胃组相比,滴鼻组诱导小鼠产生了更高水平的血清IgG和黏膜SIgA,获得了更好的免疫效果(P<0.05)。与野生型相比,密码子优化的F蛋白具有更好的免疫原性(P<0.05)。【结论】经滴鼻途径免疫和密码子优化能够提高以SL7207为载体的RSV DNA疫苗免疫效果。
【Objective】 Salmonella typhimurium aroA strain SL7207 (SL7207) was used as a carrier to carry fusion protein encoding fusion glycoprotein (F) of human respiratory syncytial vrius (RSV) Nuclear expression plasmids, to explore different mucosal immune pathway and codon optimization on the immune effect. 【Method】 The codon-optimized F gene (Fsyn) was obtained by codon-optimized Fwt gene of RSV. The eukaryotic expression plasmid pcDNA3.1 / Fsyn expressing Fsyn was constructed and SL7207 was transformed into SL7207 / pcDNA3.1 / Fsyn. The BALB / c mice were immunized by intranasal and intragastric route respectively, and the immune effect was compared by indirect ELISA. 【Results】 Compared with the intragastric administration group, the nasal drops group produced higher levels of serum IgG and mucosal SIgA and achieved better immune effect (P <0.05). The codon optimized F protein has better immunogenicity (P <0.05) compared to the wild type. 【Conclusion】 The intranasal route of immunization and codon optimization can improve the immunogenicity of RSV DNA vaccine using SL7207 as carrier.