Recombinant duck enteritis virus expressing the HA gene from goose H5 subtype avian influenza virus
Recombinant duck enteritis virus expressing the HA gene from goose H5 subtype avian influenza virus
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DOI：
10.1016/j.vaccine.2013.10.035
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发表时间：
2013-12-05
期刊：
影响因子：
5.5
通讯作者：
Wang, JunWei
中科院分区：
文献类型：
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作者：
Liu, Xiaomei;Wei, Shuangshi;Wang, JunWei
鸭肠炎病毒(DEV)基因组大，宿主范围窄，有望成为水产禽类疫苗免疫的候选病毒载体，可抵抗多种病原体。最近，我们描述了两个包含病毒US2或glgE基因缺失的DEV重组体。将鸡源H5N1型高致病性禽流感病毒(HPAIV)的血凝素(HA)基因插入缺失区，构建了表达HA抗原的rDEV。将所构建的rDEV-Delta GigE-HA或rDEV-Delta US2-HA重组DEV病毒感染鸭胚胎成纤维细胞。逆转录聚合酶链式反应、免疫荧光和免疫印迹分析结果表明，rDEV-Delta GigE-HA和rDEV-Delta US2-HA在鸭胚成纤维细胞中成功表达。为了研究HA基因在重组病毒中的稳定性，将重组病毒在DEFS中传代18次。两个重组子均可通过PCR扩增出HA基因。免疫4周龄的雏鸭能诱导出针对DEV和AIV HA的特异性抗体，并能抵抗DEV AV1221病毒的攻击感染。(C)2013爱思唯尔有限公司。保留所有权利。
The duck enteritis virus (DEV) may be a promising candidate viral vector for an aquatic poultry vaccination that can protect against multiple pathogens because it has a very large genome and a narrow host range. Recently, we described two DEV recombinants that contained deletions of the viral US2 or glgE genes. The hemagglutinin (HA) gene of an H5N1-type highly pathogenic avian influenza virus (HPAIV) of goose origin was inserted into the deletion sites to construct two rDEVs expressing the AIV HA antigen. The resulting rDEV-Delta gIgE-HA or rDEV-Delta US2-HA recombinant DEV viruses were used to infect duck embryo fibroblasts. Reverse transcription PCR, immunofluorescence and western blot analysis results indicated that rDEV-Delta gIgE-HA and rDEV-Delta US2-HA were successfully expressed in duck embryo fibroblasts (DEFs). To investigate whether the HA gene could be stably maintained in the recombinant viruses, the viruses were passaged in DEFs 18 times. The HA gene in both recombinants could be detected by PCR amplification. The immunized four-week-old ducks induced specific antibodies against DEV and AIV HA and were protected against challenge infections with DEV AV1221 viruses. (C) 2013 Elsevier Ltd. All rights reserved.