Li Zewei, Chen Yifei, Xie Haiping, Li Quan, Wang Shifeng, Shi Huoying
College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 Jiangsu China; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611-0880, USA.
Poult Sci. 2024 Dec;103(12):104436. doi: 10.1016/j.psj.2024.104436. Epub 2024 Oct 13.
Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) has resulted in significant losses for the poultry industry worldwide. Currently, there is no widely promoted vaccine for NE. In this study, immunoprecipitation (IP) was employed to isolate immunogenic proteins of C. perfringens, and 118 potential candidate antigens were identified through liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). From these, three candidate antigen proteins were selected based on their predicted antigenicity, hydrophilicity, stability, and transmembrane signalling properties: ArcB (an ornithine aminotransferase), TmpC (a probable membrane lipoprotein), and EntB (a possible enterotoxin). These three proteins were successfully produced in large quantities using Escherichia coli (E. coli), with confirmed good solubility. Both in vitro and in vivo research demonstrated that these antigens possess strong immunogenicity, eliciting robust antigen-specific humoral and cellular immune responses in chickens and mitigating NE symptoms caused by C. perfringens. The candidate antigens identified through immunoproteomics hold potential as subunit vaccines against C. perfringens infection.
由产气荚膜梭菌引起的坏死性肠炎(NE)给全球家禽业造成了重大损失。目前,尚无广泛推广的NE疫苗。在本研究中,采用免疫沉淀(IP)法分离产气荚膜梭菌的免疫原性蛋白,并通过液相色谱-质谱/质谱(LC-MS/MS)鉴定出118种潜在的候选抗原。基于预测的抗原性、亲水性、稳定性和跨膜信号特性,从这些抗原中选择了三种候选抗原蛋白:ArcB(鸟氨酸转氨酶)、TmpC(可能的膜脂蛋白)和EntB(可能的肠毒素)。利用大肠杆菌(E. coli)成功大量生产了这三种蛋白,并证实其溶解性良好。体外和体内研究均表明,这些抗原具有很强的免疫原性,能在鸡体内引发强烈的抗原特异性体液免疫和细胞免疫反应,并减轻产气荚膜梭菌引起的NE症状。通过免疫蛋白质组学鉴定出的候选抗原有望作为抗产气荚膜梭菌感染的亚单位疫苗。