Construction and immunogenicity of recombinant adenovirus vaccines expressing the HMW1, HMW2, or Hia adhesion protein of nontypeable Haemophilus influenzae.
Construction and immunogenicity of recombinant adenovirus vaccines expressing the HMW1, HMW2, or Hia adhesion protein of nontypeable Haemophilus influenzae.
复制标题
表达不可分型流感嗜血杆菌 HMW1、HMW2 或 Hia 粘附蛋白的重组腺病毒疫苗的构建和免疫原性。
DOI：
10.1128/cvi.00115-10
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Barenkamp,StephenJ
中科院分区：
文献类型：
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作者：
Winter,LindaE;Barenkamp,StephenJ
本研究的目的是构建表达流感嗜血杆菌（NTHi）HMW 1、HMW 2和Hia蛋白的重组腺病毒载体，并评价其免疫原性。这些蛋白是NTHi表达的关键粘附素和潜在的保护性抗原。将编码HMW 1或HMW 2远端一半的hmw 1A和hmw 2A结构基因片段克隆到T7表达载体pGEMEX-2中。这些构建体编码稳定的HMW 1或HMW 2重组融合蛋白，其表达大多数NTHi菌株共有的B细胞表位。还将编码成熟Hia的表面暴露部分的hiagene片段克隆到pGEMEX-2中。从亲本质粒上切下所得的T7基因10翻译融合体，并克隆到穿梭质粒pDC 316中。用pDC 316衍生物和pBHGloxΔE1，3Cre共转染HEK 293细胞导致产生病毒空斑，从中回收表达融合蛋白的重组腺病毒。用单次108-PFU剂量的HMW 2或Hia腺病毒构建体腹膜内免疫的龙猫在免疫后4周内产生高的抗HMW 2或抗Hia血清抗体滴度。用107- 109-PFU剂量的Hia腺病毒构建体鼻内免疫的龙猫在免疫后8周内也产生了高的抗Hia血清抗体滴度。重组腺病毒代表了一个有前途的系统，以诱导粘膜和全身免疫和保护粘膜疾病，如中耳炎。表达重组HMW 1、HMW 2或Hia蛋白的重组腺病毒将是NTHi疫苗开发工作中重要的新工具。
The objective of the present study was to construct and assess the immunogenicity of recombinant adenovirus vectors expressing the HMW1, HMW2, or Hia protein of nontypeableHaemophilus influenzae(NTHi). These proteins are critical adhesins and potential protective antigens expressed by NTHi. Segments of thehmw1Aandhmw2Astructural genes that encode the distal one-half of mature HMW1 or HMW2 were cloned into the T7 expression vector pGEMEX-2. These constructs encoded stable HMW1 or HMW2 recombinant fusion protein that expresses B-cell epitopes common to most NTHi strains. A segment of thehiagene that encodes the surface-exposed portion of mature Hia was also cloned into pGEMEX-2. The resulting T7 gene 10 translational fusions were excised from the parent plasmids and cloned into the shuttle plasmid pDC316. Cotransfection of HEK 293 cells with the pDC316 derivatives and pBHGloxΔE1,3Cre resulted in the production of viral plaques from which recombinant adenoviruses expressing fusion proteins were recovered. Chinchillas immunized intraperitoneally with a single 108-PFU dose of either the HMW2 or Hia adenoviral construct developed high anti-HMW2 or anti-Hia serum antibody titers within 4 weeks of immunization. Chinchillas immunized intranasally with a single 107- to 109-PFU dose of the Hia adenoviral construct also developed high anti-Hia serum antibody titers within 8 weeks of immunization. Recombinant adenoviruses represent a promising system to induce mucosal and systemic immunity and protection against mucosal diseases such as otitis media. Recombinant adenoviruses expressing recombinant HMW1, HMW2, or Hia protein will be important new tools in NTHi vaccine development efforts.