Yang Shanshan, Cao Qiuxia, Yan Kexin, Wang Chuanhong, Song Xu, Bian Xianyu, Li Sufen, Cheng Zhenkong, Zhang Xuehan, Wang Yi, Guo Rongli, Wang Xiaodu, Song Houhui, Fan Baochao, Li Bin
Key Laboratory of Veterinary Biological Engineering and Technology, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu, China; Jiangsu Key Laboratory for Food Quality and Safety-State, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Ministry of Agriculture, Nanjing, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, PR China.
Key Laboratory of Veterinary Biological Engineering and Technology, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu, China; College of Veterinary Medicine, Zhejiang A&F University, Hangzhou 311300, Zhejiang, PR China.
Cytokine. 2025 Apr;188:156880. doi: 10.1016/j.cyto.2025.156880. Epub 2025 Feb 7.
The insufficiency of current Porcine Epidemic Diarrhea (PED) vaccines against highly pathogenic strains highlights the critical importance of enhancing mucosal immunity in the prevention and control of porcine enteric viral diseases. Due to limited research platforms, the understanding of the porcine mucosal immune system and its response mechanisms remains incomplete. This study employed prokaryotic expression and purification methods to obtain eight essential cytokines involved in mucosal immune responses (CD40L, IL-2, IL-6, TNF-α, IL-13, IL-17α, TGF-β, APRIL). By utilizing various cell models including porcine intestinal organoids, IPEC-J2, Vero-E6, porcine peripheral blood lymphocytes, and porcine Peyer's patch lymphocytes, the functions of these eight cytokines were identified through flow cytometry, immunoblotting, relative quantitative PCR, and CFSE proliferation assays. The results demonstrate that all eight purified proteins exhibit both protein activity and function. The purification of these molecules lays the groundwork for further exploration of the mucosal barrier of pigs and mucosal immune-related studies, as well as providing research tools for the prevention and control of enteric viral diseases in pigs.
当前猪流行性腹泻（PED）疫苗对高致病性毒株的效力不足，这凸显了增强黏膜免疫在猪肠道病毒性疾病防控中的至关重要性。由于研究平台有限，对猪黏膜免疫系统及其反应机制的了解仍不完整。本研究采用原核表达和纯化方法，获得了参与黏膜免疫反应的八种关键细胞因子（CD40L、IL-2、IL-6、TNF-α、IL-13、IL-17α、TGF-β、APRIL）。通过利用包括猪肠道类器官、IPEC-J2、Vero-E6、猪外周血淋巴细胞和猪派伊尔结淋巴细胞在内的多种细胞模型，通过流式细胞术、免疫印迹、相对定量PCR和CFSE增殖试验确定了这八种细胞因子的功能。结果表明，所有八种纯化蛋白均具有蛋白活性和功能。这些分子的纯化，为进一步探索猪的黏膜屏障和黏膜免疫相关研究奠定了基础，也为猪肠道病毒性疾病的防控提供了研究工具。
