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用于猪传染性胸膜肺炎广谱免疫保护的重组galT-galU蛋白的研制与特性分析

Development and Characterization of a Recombinant galT-galU Protein for Broad-Spectrum Immunoprotection Against Porcine Contagious Pleuropneumonia.

作者信息

Chen Jia-Yong, Deng Yi, Liu Jiale, Wen Xin, Cao Yu-Qin, Mu Yu, Sun Mengke, Miao Chang, Peng Zhiling, Lu Kun, Wang Yu-Luo, Chen Xizhu, Pang Siyu, Wang Dan, Zhou Jiayu, Li Miaohan, Wen Yiping, Wu Rui, Zhao Shan, Lang Yi-Fei, Yan Qi-Gui, Huang Xiaobo, Du Senyan, Wang Yiping, Han Xinfeng, Cao San-Jie, Zhao Qin

机构信息

Research Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.

Sichuan Science-Observation Experimental Station of Veterinary Drugs and Veterinary Biotechnology, Ministry of Agriculture and Rural Affairs, Chengdu 611130, China.

出版信息

Int J Mol Sci. 2025 Apr 11;26(8):3634. doi: 10.3390/ijms26083634.

Abstract

Porcine contagious pleuropneumonia (PCP), caused by (APP), is a highly contagious disease that leads to significant economic losses in the swine industry. Current vaccines are ineffective due to the presence of multiple serotypes and the absence of a predominant seasonal serotype, underscoring the need for vaccines with broad-spectrum protection. Previous studies identified galT and galU as promising antigen candidates. In this study, we expressed and characterized a soluble recombinant galT-galU protein (rgalT-galU) from the pET-28a-galT-galU plasmid. The protein, with a molecular weight of 73 kDa, exhibited pronounced immunogenicity in murine models, as indicated by a significant elevation in IgG titers determined through an indirect ELISA. This immune response was further corroborated by substantial antigen-specific splenic lymphocyte proliferation, with a stimulation index of 51.5%. Immunization also resulted in elevated serum cytokines levels of IL-4, IL-12, and IFN-γ, as detected by cytokine assays. Vaccination with rgalT-galU provided immunoprotection against three predominant APP strains (APP1, APP5b, and APP7), achieving protection rates of 71.4%, 71.4%, and 85.7%, respectively. It also effectively mitigated pulmonary lesions and neutrophil infiltration, as verified by histopathological and immunohistochemical analyses. These results indicate that rgalT-galU is a promising candidate for developing cross-protective subunit vaccines against APP infection.

摘要

猪传染性胸膜肺炎(PCP)由胸膜肺炎放线杆菌(APP)引起,是一种高度传染性疾病,给养猪业造成重大经济损失。由于存在多种血清型且缺乏占主导地位的季节性血清型,目前的疫苗效果不佳,这凸显了对具有广谱保护作用疫苗的需求。先前的研究确定galT和galU为有前景的抗原候选物。在本研究中,我们从pET-28a-galT-galU质粒中表达并鉴定了一种可溶性重组galT-galU蛋白(rgalT-galU)。该蛋白分子量为73 kDa,在小鼠模型中表现出显著的免疫原性,通过间接ELISA测定的IgG滴度显著升高表明了这一点。大量的抗原特异性脾淋巴细胞增殖进一步证实了这种免疫反应,刺激指数为51.5%。免疫接种还导致通过细胞因子检测发现血清中IL-4、IL-12和IFN-γ的细胞因子水平升高。用rgalT-galU进行疫苗接种对三种主要的APP菌株(APP1、APP5b和APP7)提供了免疫保护,保护率分别达到71.4%、71.4%和85.7%。组织病理学和免疫组织化学分析证实,它还有效减轻了肺部病变和中性粒细胞浸润。这些结果表明,rgalT-galU是开发针对APP感染的交叉保护亚单位疫苗的有前景的候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de62/12027175/0429f81713e3/ijms-26-03634-g001.jpg

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