以减毒鼠伤寒沙门菌为载体的人呼吸道合胞病毒F蛋白DNA疫苗滴鼻免疫抗体分析  

Antibody responses induced by mucosal DNA vaccine encoding the codon-optimized F protein of human respiratory syncytial virus (RSV) delivered with attenuated Salmonella typhimurium

作　　者：程明[1,2] 何金生[2] 付远辉[2] 乔伟[1] 焦月盈[2] 

机构地区：[1]安徽医科大学免疫学教研室,合肥230032 [2]北京交通大学生命科学与生物工程研究院,北京100044

出　　处：《微生物学报》2011年第7期965-971,共7页Acta Microbiologica Sinica

基　　金：北京市自然科学基金(7092053)~~

摘　　要：【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] In order to investigate antibody responses by musosal DNA vaccines encoding the codon-optimized F protein of human respiratory syncytial virus（RSV） delivered with attenuated Salmonella typhimurium aroA strain SL7207（SL7207） via intranasal or intragastric routes.[Methods] After the codon-optimized F gene was synthesized,we constructed eukaryotic expression plasmid pcDNA3.1/Fsyn and transformed it into SL7207 to get recombinant SL7207 of SL7207/pcDNA3.1/Fsyn.Then,SL7207/pcDNA3.1/Fsyn was exploited to evaluate its immunogenicity in BALB/c mice administered intranasally or intragastrically.The resultant serum and mucosal antibody responses were analyzed by indirect ELISA.[Results] Compared with intragastric immunization group,intranasal immunization group achieved higher level of RSV specific serum IgG and secretion IgA（SIgA）（P0.05）.Moreover,F protein expressed by the codon-optimized F gene of Fsyn was of elevated immunogenicity,compared with wild type F（Fwt）（P0.05）.[Conclusion] Intranasal immunization and codon optimization approaches can improve antibody responses of RSV DNA vaccine delivered by attenuated Salmonella typhimurium aroA strain SL7207.

关 键 词：人呼吸道合胞病毒 F蛋白密码子优化 黏膜免疫 减毒鼠伤寒沙门氏菌SL7207 

分 类 号：R392[医药卫生—免疫学]