Department of Genetic Medicine, Weill Cornell Medical College , New York, NY 10026, USA.
Hum Gene Ther. 2010 Jul;21(7):891-901. doi: 10.1089/hum.2009.148.
The aerosol form of the bacterium Yersinia pestis causes pneumonic plague, a rapidly fatal disease that is a biothreat if deliberately released. At present, no plague vaccines are available for use in the United States, but subunit vaccines based on the Y. pestis V antigen and F1 capsular protein show promise when administered with adjuvants. In the context that adenovirus (Ad) gene transfer vectors have a strong adjuvant potential related to the ability to directly infect dendritic cells, we hypothesized that modification of the Ad5 capsid to display either the Y. pestis V antigen or the F1 capsular antigen on the virion surface would elicit high V antigen- or F1-specific antibody titers, permit boosting with the same Ad serotype, and provide better protection against a lethal Y. pestis challenge than immunization with equivalent amounts of V or F1 recombinant protein plus conventional adjuvant. We constructed AdYFP-pIX/V and AdLacZ-pIX/F1, E1(-), E3(-) serotype 5 Ad gene transfer vectors containing a fusion of the sequence for either the Y. pestis V antigen or the F1 capsular antigen to the carboxy-terminal sequence of pIX, a capsid protein that can accommodate the entire V antigen (37 kDa) or F1 protein (15 kDa) without disturbing Ad function. Immunization with AdYFP-pIX/V followed by a single repeat administration of the same vector at the same dose resulted in significantly better protection of immunized animals compared with immunization with a molar equivalent amount of purified recombinant V antigen plus Alhydrogel adjuvant. Similarly, immunization with AdLacZ-pIX/F1 in a prime-boost regimen resulted in significantly enhanced protection of immunized animals compared with immunization with a molar-equivalent amount of purified recombinant F1 protein plus adjuvant. These observations demonstrate that Ad vaccine vectors containing pathogen-specific antigens fused to the pIX capsid protein have strong adjuvant properties and stimulate more robust protective immune responses than equivalent recombinant protein-based subunit vaccines administered with conventional adjuvant, suggesting that F1-and/or V-modified capsid Ad-based recombinant vaccines should be considered for development as anti-plague vaccines.
鼠疫耶尔森菌的气溶胶形式引起肺鼠疫,这是一种迅速致命的疾病,如果故意释放,将构成生物威胁。目前,美国没有可用的鼠疫疫苗,但基于鼠疫耶尔森菌 V 抗原和 F1 荚膜蛋白的亚单位疫苗在与佐剂一起使用时显示出前景。在腺病毒 (Ad) 基因转移载体具有与直接感染树突状细胞的能力相关的强大佐剂潜力的背景下,我们假设修饰 Ad5 衣壳以在病毒表面上显示鼠疫耶尔森菌 V 抗原或 F1 荚膜抗原将引起高 V 抗原或 F1 特异性抗体滴度,允许用相同的 Ad 血清型进行加强,并且比用相同量的 V 或 F1 重组蛋白加常规佐剂免疫能更好地保护免受致死性鼠疫耶尔森菌的挑战。我们构建了 AdYFP-pIX/V 和 AdLacZ-pIX/F1、E1(-)、E3(-) 血清型 5 Ad 基因转移载体,其中包含 V 抗原或 F1 荚膜抗原的序列与 pIX 的羧基末端序列的融合,pIX 是一种衣壳蛋白,可容纳整个 V 抗原(37 kDa)或 F1 蛋白(15 kDa)而不干扰 Ad 功能。用 AdYFP-pIX/V 免疫,然后用相同剂量的同一载体重复一次,与用等量纯化的重组 V 抗原加 Alhydrogel 佐剂免疫的动物相比,显著更好地保护了免疫动物。同样,用 AdLacZ-pIX/F1 进行初免-加强免疫方案,与用等量纯化的重组 F1 蛋白加佐剂免疫的动物相比,显著增强了免疫动物的保护作用。这些观察结果表明,含有与 pIX 衣壳蛋白融合的病原体特异性抗原的 Ad 疫苗载体具有强大的佐剂特性,并刺激比用常规佐剂给予的等效重组蛋白亚单位疫苗更强大的保护性免疫应答,表明 F1-和/或 V 修饰的衣壳 Ad 基于重组疫苗应被视为抗鼠疫疫苗的发展。