Intranasal immunization with single-dose vaccine based on recombinant influenza virus H1N1 expressing the extracellular domain of respiratory syncytial virus G protein induces robust immunity and protection in mice
Intranasal immunization with single-dose vaccine based on recombinant influenza virus H1N1 expressing the extracellular domain of respiratory syncytial virus G protein induces robust immunity and protection in mice
目的 基于重组流感病毒载体的呼吸道合胞病毒(RSV)疫苗的构建及在小鼠体内的免疫保护效果评价。 方法 构建并拯救表达RSV A2型G蛋白胞外结构域(Gecto)的重组甲型流感病毒，将其命名为PR8NAGecto/WSN。体外验证重组病毒G蛋白表达与病毒生长动力学后，单剂滴鼻免疫BALB/c小鼠，并评价体液免疫、黏膜免疫与细胞免疫。免疫4周后，分别用RSV A2与RSV B9320进行攻毒，通过小鼠体重变化、肺组织病毒滴度及病理评价免疫保护效果。 结果 单剂滴鼻免疫PR8NAGecto/WSN能在小鼠体内产生较强的体液免疫、黏膜免疫及细胞免疫。与对照组比较，免疫组小鼠经RSV A2或B9320两个亚型病毒攻毒后肺病毒载量与肺组织病理均明显改善。 结论 单剂滴鼻免疫重组PR8NAGecto/WSN疫苗可在小鼠体内诱导较强的RSV特异免疫应答与攻毒保护。本研究为新型RSV黏膜疫苗的研发提供新的思路与实验资料。 Objective To construct a novel respiratory syncytial virus (RSV) vaccine based on a recombinant influenza virus vector and evaluate its immune protective effects in mice. Methods A recombinant H1N1 influenza A virus (IAV) expressing the extracellular domain (Gecto) of RSV A2 G protein was constructed and rescued, named as PR8NAGecto/WSN. After in vitro verification of the Gecto expression and PR8NAGecto/WSN growth kinetics, a single dose of PR8NAGecto/WSN was used to immunize BALB/c mice through intranasal administration to evaluate the efficacy of PR8NAGecto/WSN by assessing humoral (IgG, neutralizing antibody), mucosal (IgA) and cellular immunity (IFN-γ ELISPOT). Four weeks after immunization, the mice were challenged with RSV A2 or RSV B9320 to evaluate the protective effects of PR8NAGecto/WSN by analyzing mouse body weight changes, lung tissue virus titers and pathological changes. Results A single-dose intranasal immunization with PR8NAGecto/WSN induced robust humoral, mucosal and cellular immunity in mice. Moreover, the mice in the immunized group had lower lung virus loads and mild lung pathological damages following the challenge with RSV A or RSV B subtype as compared with the control group. Conclusions A single-dose intranasal immunization with PR8NAGecto/WSN induces robust immunity and provide protection against RSV A and B challenges in mice. This study provides new ideas and reference for the development of novel mucosal vaccines against RSV.
Abstract
Objective To construct a novel respiratory syncytial virus (RSV) vaccine based on a recombinant influenza virus vector and evaluate its immune protective effects in mice. Methods A recombinant H1N1 influenza A virus (IAV) expressing the extracellular domain (Gecto) of RSV A2 G protein was constructed and rescued, named as PR8NAGecto/WSN. After in vitro verification of the Gecto expression and PR8NAGecto/WSN growth kinetics, a single dose of PR8NAGecto/WSN was used to immunize BALB/c mice through intranasal administration to evaluate the efficacy of PR8NAGecto/WSN by assessing humoral (IgG, neutralizing antibody), mucosal (IgA) and cellular immunity (IFN-γ ELISPOT). Four weeks after immunization, the mice were challenged with RSV A2 or RSV B9320 to evaluate the protective effects of PR8NAGecto/WSN by analyzing mouse body weight changes, lung tissue virus titers and pathological changes. Results A single-dose intranasal immunization with PR8NAGecto/WSN induced robust humoral, mucosal and cellular immunity in mice. Moreover, the mice in the immunized group had lower lung virus loads and mild lung pathological damages following the challenge with RSV A or RSV B subtype as compared with the control group. Conclusions A single-dose intranasal immunization with PR8NAGecto/WSN induces robust immunity and provide protection against RSV A and B challenges in mice. This study provides new ideas and reference for the development of novel mucosal vaccines against RSV.
关键词
呼吸道合胞病毒/流感病毒/病毒载体/G蛋白
Key words
Respiratory syncytial virus/Influenza virus/Virus vector/G attachment protein
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