Wang Jialing, Chen Yan, He Xiaohua, Du Xiaoli, Gao Yongheng, Shan Xinggen, Hu Zhiqun, Hu Qinghai
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
Front Microbiol. 2023 Jan 11;13:1082712. doi: 10.3389/fmicb.2022.1082712. eCollection 2022.
mainly infects domestic ducks, geese, turkeys, and other birds, and causes considerable economic losses to the global duck industry. Previous studies have shown that concentrated cell-free culture filtrates of induce highly significant protection against homologous challenge. In this study, 12 immunogenic proteins were identified in the culture supernatant of strain Yb2 with immunoproteomic analysis. Of these, three immunogenic proteins, AS87_RS06600 (designated "PaR1" in this study), AS87_RS09020, and AS87_RS09965, which appeared in more than three spots on the western-blotted membrane, were expressed in and purified. Animal experiments showed that the recombinant PaR1 (rPaR1) protein protected 41.67% of immunized ducklings against challenge with virulent Yb2, whereas rAS87_RS09020 or rAS87_RS09965 did not, and that ducklings immunized once with rPaR1 were 20, 40, and 0% protected from challenge with strains WJ4 (serotype 1), Yb2 (serotype 2), and HXb2 (serotype 10), respectively. In addition, rPaR1 immunized rabbit serum showed bactericidal activity against strain Yb2 at a titer of 1:8. These results indicate that rPaR1 of strain Yb2 protects against homologous challenge. Amino acid homology analysis show that PaR1 is a non-serotype-specific protein among different serotypes. Furthermore, PaR1 is mainly secreted outside the cell through the T9SS. Overall, our results demonstrate that PaR1 is a non-serotype-specific protective protein secreted by the T9SS.
主要感染家鸭、鹅、火鸡及其他禽类,给全球鸭产业造成巨大经济损失。先前研究表明,[病原体名称]的浓缩无细胞培养滤液对同源攻击具有高度显著的保护作用。在本研究中,通过免疫蛋白质组学分析在[病原体名称]菌株Yb2的培养上清液中鉴定出12种免疫原性蛋白。其中,在western印迹膜上出现超过三个斑点的三种免疫原性蛋白,即AS87_RS06600(本研究中命名为“PaR1”)、AS87_RS09020和AS87_RS09965,在[表达系统名称]中表达并纯化。动物实验表明,重组PaR1(rPaR1)蛋白可保护41.67%的免疫雏鸭免受强毒株Yb2的攻击,而rAS87_RS09020或rAS87_RS09965则不能,并且用rPaR1免疫一次的雏鸭分别对WJ4(血清型1)、Yb2(血清型2)和HXb2(血清型10)菌株攻击的保护率为20%、40%和0%。此外,rPaR1免疫兔血清对菌株Yb2的杀菌效价为1:8。这些结果表明,菌株Yb2的rPaR1可抵御同源攻击。氨基酸同源性分析表明,PaR1是不同[病原体名称]血清型之间的非血清型特异性蛋白。此外,PaR1主要通过T9分泌系统分泌到细胞外。总体而言,我们的结果表明,[病原体名称]PaR1是一种由T9分泌系统分泌的非血清型特异性保护蛋白。