Shahriari Amir Ghaffar, Bagheri Abdolreza, Afsharifar Alireza, Habibi-Pirkoohi Maziar
Department of Agriculture and Natural Resources, Higher Education Center of Eghlid, Eghlid, Iran.
Department of Crop Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, Iran.
Iran J Biotechnol. 2019 Jan 11;17(1):e2215. doi: 10.21859/ijb.2215. eCollection 2019 Jan.
Newcastle disease is a major avian disease that causes enormous economic loss in poultry industry. There have been a number of reports on the suitability of plant-based recombinant vaccine against this disease. Fusion (F) and hemagglutinin-neuraminidase (HN) epitopes of the Newcastle disease virus (NDV) represent the major immunogenic sites for development of recombinant anti-ND vaccines in plant hosts.
The main objective of this research was to evaluate the ability of a recombinant anti-ND vaccine in induction of immune responses in animal model.
In this study, immunogenicity of recombinant fusion (F) and hemagglutinin-neuraminidase (HN) epitopes of the Newcastle disease virus (NDV) is investigated in an animal model. The corresponding genes encoding amino acids 65-81 of the F protein and 346-353 amino acids of HN were expressed in tobacco seedling using agrobacterium-mediated transformation. Expression of the foreign gene in the tobacco seedlings was investigated by a number of molecular assays including Real-Time PCR and ELISA. Transgenic plant extract was used to induce immunogenic response in animal model.
Integration of the foreign gene in plant host genome was confirmed by polymerase chain reaction (PCR). Expression of the foreign recombinant protein was confirmed by Real-Time PCR and ELISA assays. Immunogenicity of the recombinant protein was investigated in rabbit by subcutaneous injection. Results indicated that the transgenic plant extract can induce immune responses in the host as confirmed by presence of specific antibodies in the sera in ELISA assay. Western blot assays showed that the foreign gene was actually expressed in transgenic seedlings.
The results obtained in this research provide further evidence on applicability of plant-based recombinant vaccines for protection of poultry against Newcastle disease.
新城疫是一种主要的禽类疾病，给家禽业造成巨大经济损失。关于植物源重组疫苗针对该疾病的适用性已有多篇报道。新城疫病毒（NDV）的融合（F）和血凝素 - 神经氨酸酶（HN）表位是在植物宿主中开发重组抗新城疫疫苗的主要免疫原性位点。
本研究的主要目的是评估重组抗新城疫疫苗在动物模型中诱导免疫反应的能力。
在本研究中，在动物模型中研究新城疫病毒（NDV）重组融合（F）和血凝素 - 神经氨酸酶（HN）表位的免疫原性。使用农杆菌介导的转化在烟草幼苗中表达编码F蛋白65 - 81位氨基酸和HN 346 - 353位氨基酸的相应基因。通过包括实时PCR和ELISA在内的多种分子检测方法研究外源基因在烟草幼苗中的表达。转基因植物提取物用于在动物模型中诱导免疫原性反应。
通过聚合酶链反应（PCR）确认外源基因整合到植物宿主基因组中。通过实时PCR和ELISA检测确认外源重组蛋白的表达。通过皮下注射在兔中研究重组蛋白的免疫原性。结果表明，ELISA检测血清中存在特异性抗体证实转基因植物提取物可在宿主中诱导免疫反应。蛋白质印迹分析表明外源基因确实在转基因幼苗中表达。
本研究获得的结果为植物源重组疫苗用于保护家禽免受新城疫侵害的适用性提供了进一步证据。
