Generation of influenza virus from avian cells infected by Salmonella carrying the viral genome.
Generation of influenza virus from avian cells infected by Salmonella carrying the viral genome.
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DOI：
10.1371/journal.pone.0119041
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Curtiss R 3rd
中科院分区：
文献类型：
--
作者：
Zhang X;Kong W;Wanda SY;Xin W;Alamuri P;Curtiss R 3rd
家禽是将甲型流感病毒从野生水禽水库传播给人类的中间媒介，导致禽流感在家禽中爆发，并可能在人类中流行。为了对抗新出现的禽流感病毒，一种廉价的、热稳定的口服流感疫苗将有助于为大型商业家禽甚至候鸟接种疫苗。我们假设的疫苗是一种重组减毒菌株，能够在体内介导流感病毒的减毒，从而诱导对流感的保护性免疫。在此，我们报告了基于探索性研究的这种理想疫苗的可行性和技术限制。构建了5个携带氯霉素耐药基因或不含抗生素耐药标记的8单元质粒。这两种质粒分别转染后，在禽细胞中成功地产生了流感病毒。沙门氏菌载体被设计成能够稳定维持并有条件地将8个单位的质粒释放到禽流感细胞中，以恢复流感病毒。在26例鸡胚成纤维细胞（CEF）和Madin-Darby犬肾（MDCK）细胞共培养的11例中，携带8单位质粒的重组沙门氏菌感染甲型流感病毒，其组织培养感染剂量（TCID50）高达107 50% /ml。我们的数据证明，细菌载体可以通过将其DNA货物运送到允许的宿主细胞中来介导流感病毒的产生。虽然我们在开发这种流感沙门氏菌疫苗递送系统方面取得了进展，但要实现有效的病毒生产，特别是在体内，还需要进一步改进。
Domestic poultry serve as intermediates for transmission of influenza A virus from the wild aquatic bird reservoir to humans, resulting in influenza outbreaks in poultry and potential epidemics/pandemics among human beings. To combat emerging avian influenza virus, an inexpensive, heat-stable, and orally administered influenza vaccine would be useful to vaccinate large commercial poultry flocks and even migratory birds. Our hypothesized vaccine is a recombinant attenuated bacterial strain able to mediate production of attenuated influenza virus in vivo to induce protective immunity against influenza. Here we report the feasibility and technical limitations toward such an ideal vaccine based on our exploratory study. Five 8-unit plasmids carrying a chloramphenicol resistance gene or free of an antibiotic resistance marker were constructed. Influenza virus was successfully generated in avian cells transfected by each of the plasmids. The Salmonella carrier was engineered to allow stable maintenance and conditional release of the 8-unit plasmid into the avian cells for recovery of influenza virus. Influenza A virus up to 107 50% tissue culture infective doses (TCID50)/ml were recovered from 11 out of 26 co-cultures of chicken embryonic fibroblasts (CEF) and Madin-Darby canine kidney (MDCK) cells upon infection by the recombinant Salmonella carrying the 8-unit plasmid. Our data prove that a bacterial carrier can mediate generation of influenza virus by delivering its DNA cargoes into permissive host cells. Although we have made progress in developing this Salmonella influenza virus vaccine delivery system, further improvements are necessary to achieve efficient virus production, especially in vivo.
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