Motamedi Mohammad Javad, Shahsavandi Shahla, Amani Jafar, Kazemi Rouhollah, Takrim Somayeh, Jafari Mahyat, Salmanian Ali-Hatef
Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization, Karaj, Iran.
Iran J Biotechnol. 2018 Dec 12;16(4):e1749. doi: 10.21859/ijb.1749. eCollection 2018 Dec.
Newcastle disease virus (NDV) is a dangerous viral disease, infecting a broad range of birds, and has a fatal effect on the poultry industries. The attachment and consequently fusion of the virus to the host cell membrane is directed by the two superficial glycoproteins, the hemagglutinin-neuraminidase (HN) and the fusion (F) which is considered as the important targets for the poultry immune response.
The principal goal of this investigation was to realize the potential efficacy of the expression system for the production of the multi-epitopic HN, and F proteins with respect to the ability for the stimulation of the immune system and production of the cross-reactive antibodies in mice.
The recombinant HN and F (rHN, rF) have accumulated almost 40% of the total bacterial proteins. The presence of rHN and rF proteins recognized by the Western blotting with specific anti-HN, anti-F, anti-Newcastle B1, and anti-poly 6x His-tag antibodies. Furthermore, both rHN and rF have shown the specific reactivity against the Newcastle B1 antiserum as a standard strain.
The ELISA analysis showed that the higher dilutions of the antibody against Newcastle B1 could react with the as least quantity as 100 ng of the purified rHN, and rF. Cross-reactivity analysis of the sera from the mice immunized with Newcastle B1 in two time points indicated that the raise of anti-Newcastle B1, anti-HN and anti-F antibodies peaked at 28 days post immunization (dpi). Moreover, temporal variation in IgG titration between both time points was significant at 5% probability level.
The results provided valuable information about the cross-reactivity patterns and biological activity of the multi-epitopic proteins compared to the NDV standard strain which was determined by the Western blotting and ELISA.
新城疫病毒（NDV）是一种危险的病毒性疾病，可感染多种禽类，对家禽业具有致命影响。病毒与宿主细胞膜的附着及随后的融合由两种表面糖蛋白介导，即血凝素神经氨酸酶（HN）和融合蛋白（F），它们被认为是家禽免疫反应的重要靶点。
本研究的主要目标是了解表达系统生产多表位HN和F蛋白在刺激小鼠免疫系统及产生交叉反应抗体方面的潜在功效。
重组HN和F（rHN、rF）积累量几乎占细菌总蛋白的40%。用特异性抗HN、抗F、抗新城疫B1和抗多聚6x组氨酸标签抗体进行蛋白质印迹法可识别rHN和rF蛋白的存在。此外，rHN和rF对作为标准毒株的新城疫B1抗血清均表现出特异性反应。
ELISA分析表明，抗新城疫B1抗体的较高稀释度能与低至100 ng的纯化rHN和rF发生反应。对在两个时间点用新城疫B1免疫的小鼠血清进行交叉反应分析表明，抗新城疫B1、抗HN和抗F抗体在免疫后28天达到峰值。此外，两个时间点之间IgG滴定度的时间变化在5%概率水平上具有显著性。
结果提供了关于多表位蛋白与新城疫病毒标准毒株相比的交叉反应模式和生物学活性的有价值信息，这些信息通过蛋白质印迹法和ELISA得以确定。
