Mulondo Goodman, Buyse Mélie L R, Labuschagne Kimberley, Jarvis David, van Zyl Albertha, Rybicki Edward P, Hitzeroth Inga I, Mbewana Sandiswa
Department of Molecular and Cell Biology, University of Cape Town, Cape Town, Rondebosch, 7700, South Africa.
Liselo Labs, Howick, 3291, South Africa.
Arch Virol. 2025 Jun 25;170(7):163. doi: 10.1007/s00705-025-06352-z.
Beak and feather disease virus (BFDV), a single-stranded DNA virus, infects endangered psittacine species, including the South African Cape parrot. The disease is highly contagious and can be transmitted through contact with contaminated faeces, crop secretions, and feather and skin dander. To date, there is no vaccine or cure available for BFDV. The production of an effective vaccine depends on having a production platform and methods that are both easy to use and capable of yielding a significant amount of protein that will induce a sufficient immune response. Therefore, the aim of this study was to produce a plant-based BFDV vaccine candidate and to evaluate its ability to elicit an immune response in birds.Recombinant BFDV capsid protein (CP) was transiently expressed in Nicotiana benthamiana and purified using density gradient ultracentrifugation. Japanese quails were immunized with purified BFDV CP. Yolk-derived IgY was purified by water dilution and salt precipitation, and its specificity was verified by western blot analysis. The expression levels of the coat protein increased from non-detectable to an average accumulation of 1.58 mg/kg of fresh plant tissue biomass, and antibodies against BFDV CP were detected in both the blood and eggs of immunized quails, indicating that vaccination with BFDV CP successfully elicited a humoral immune response.This study demonstrates that heterologous expression in plants is a viable method for producing BFDV CP. To the best of our knowledge, this is the first study to show the antibody response to a plant-produced BFDV antigen in a quail model. Given that the presence of anti-CP antibodies in infected birds is associated with immunity, this system can potentially be used to produce a vaccine against BFDV.
喙羽病病毒(BFDV)是一种单链DNA病毒,可感染包括南非海角鹦鹉在内的濒危鹦鹉物种。该疾病具有高度传染性,可通过接触受污染的粪便、嗉囊分泌物以及羽毛和皮屑传播。迄今为止,尚无针对BFDV的疫苗或治疗方法。有效疫苗的生产取决于拥有易于使用且能够产生大量可诱导充分免疫反应的蛋白质的生产平台和方法。因此,本研究的目的是生产一种基于植物的BFDV候选疫苗,并评估其在鸟类中引发免疫反应的能力。
重组BFDV衣壳蛋白(CP)在本氏烟草中瞬时表达,并通过密度梯度超速离心进行纯化。用纯化的BFDV CP免疫日本鹌鹑。通过水稀释和盐沉淀法纯化蛋黄来源的IgY,并通过蛋白质印迹分析验证其特异性。衣壳蛋白的表达水平从不可检测增加到平均积累量为每千克新鲜植物组织生物量1.58毫克,并且在免疫鹌鹑的血液和卵中均检测到针对BFDV CP的抗体,表明用BFDV CP进行疫苗接种成功引发了体液免疫反应。
本研究表明,在植物中进行异源表达是生产BFDV CP的可行方法。据我们所知,这是第一项在鹌鹑模型中显示对植物产生的BFDV抗原产生抗体反应的研究。鉴于感染鸟类中抗CP抗体的存在与免疫力相关,该系统有可能用于生产针对BFDV的疫苗。