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首页> 外文期刊>Virus Research: An International Journal of Molecular and Cellular Virology >Recombinant adenovirus co-expressing capsid proteins of two serotypes of foot-and-mouth disease virus (FMDV): in vitro characterization and induction of neutralizing antibodies against FMDV in swine.
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Recombinant adenovirus co-expressing capsid proteins of two serotypes of foot-and-mouth disease virus (FMDV): in vitro characterization and induction of neutralizing antibodies against FMDV in swine.

机译：重组腺病毒共表达两种口蹄疫病毒（FMDV）血清型的衣壳蛋白：猪中FMDV的中和抗体的体外表征和诱导。

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摘要

Human adenovirus type 5 (Ad5) has been evaluated as a novel gene delivery vector for the development of live-viral vaccines for foot-and-mouth disease (FMD). In this study, we constructed an Ad5 vector co-expressing the capsid precursor proteins, P1, of FMD virus (FMDV) field strains A24 Cruzeiro and O1 Campos and examined the neutralizing antibody responses in swine after inoculation with the vector. To construct the Ad5 vector, a bicistronic expression cassette containing a cytomegalovirus promoter, the P1 coding sequence of FMDV A24, the internal ribosomal entry site (IRES) of FMDV A12, the P1 coding sequence of FMDV O1 Campos and the coding region of A12 3C protease was inserted into the E1 region of an E1/E3-deleted Ad5. The recombinant adenovirus, Ad5A24+O1, was generated by transfection of 293 cells with full-length pAd5A24+O1 recombinant plasmid DNA. The recombinant Ad5 co-expressed P1 of both A24 and O1 in infected 293 cells and P1 of both serotypes was processed to produce VP0, VP3, and VP1. We further demonstrated the formation of capsid protein complexes by co-precipitation of VP0, VP3, and VP1 with monoclonal antibodies against viral capsid proteins. Swine inoculated with Ad5A24+O1 generated neutralizing antibodies against both A24 and O1. However, the overall neutralizing antibody response was considerably lower than that induced by a commercial FMD vaccine or a monovalent Ad5-A24 vaccine.
机译：人类腺病毒5型（Ad5）已被评估为开发用于口蹄疫（FMD）的活病毒疫苗的新型基因传递载体。在这项研究中，我们构建了一个Ad5载体，该载体共表达FMD病毒（FMDV）田间菌株A24 Cruzeiro和O1 Campos的衣壳前体蛋白P1，并在接种该载体后检查了猪中和抗体的反应。为了构建Ad5载体，含有巨细胞病毒启动子，FMDV A24的P1编码序列，FMDV A12的内部核糖体进入位点（IRES），FMDV O1 Campos的P1编码序列以及A12 3C的编码区的双顺反子表达盒将蛋白酶插入缺失E1 / E3的Ad5的E1区域。重组腺病毒Ad5A24 + O1是通过用全长pAd5A24 + O1重组质粒DNA转染293细胞而产生的。重组的Ad5在感染的293细胞中共同表达A24和O1的P1，并处理两种血清型的P1，以产生VP0，VP3和VP1。我们进一步证明了VP0，VP3和VP1与抗病毒衣壳蛋白的单克隆抗体共沉淀可形成衣壳蛋白复合物。接种Ad5A24 + O1的猪产生针对A24和O1的中和抗体。然而，总的中和抗体应答明显低于商业FMD疫苗或单价Ad5-A24疫苗诱导的应答。

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