Xin Cheng, Chen Zhuting, Zhou Jingming, Chen Yumei, Liu Yankai, Liu Hongliang, Liang Chao, Zhu Xifang, Qi Yanhua, Zhang Gaiping, Wang Aiping
School of Life Sciences, Zhengzhou University, Henan, Zhengzhou, China.
Longhu Laboratory, Henan, Zhengzhou, China.
Microbiol Spectr. 2024 Oct 23;12(12):e0141124. doi: 10.1128/spectrum.01411-24.
African swine fever virus (ASFV) is a highly contagious and often fatal pathogen that poses a significant threat to the swine industry worldwide. The H171R protein, a structural component of ASFV, plays crucial roles in viral assembly, host cell entry, and modulation of the host immune response. This study aimed to comprehensively characterize the linear B-cell epitopes on the H171R protein to facilitate the development of diagnostic tools and subunit vaccines against ASFV. A combined approach involving bioinformatics analysis and experimental techniques was employed. The recombinant H171R protein was expressed and purified, and specific monoclonal antibodies were generated through immunization and hybridoma technology. Systematic epitope mapping using overlapping peptide fragments and alanine-scanning mutagenesis revealed four minimal linear epitopes: HPLLPYQQSSDEQP, SDEQPMMPYQQPPG, PYEQIYHKKHASQQ, and LNDYYQHILALGDED. The identified epitopes exhibited strong immunogenicity, as demonstrated by their reactivity with ASFV-positive swine sera. Critical amino acid residues within each epitope were identified through mutational analysis. Structural modeling and visualization of the H171R protein provided insights into the spatial distribution and accessibility of the epitope regions. These findings contribute to a better understanding of the H171R protein's antigenic properties and lay the foundation for developing effective diagnostic assays and subunit vaccines against African swine fever.
African swine fever virus (ASFV) poses a severe threat to the global swine industry. This study characterizes linear B-cell epitopes on the crucial ASFV H171R protein, facilitating the development of improved diagnostics and subunit vaccines. Four immunogenic epitopes were identified, offering valuable information for designing sensitive diagnostic assays and potential subunit vaccine candidates. By advancing the understanding of H171R's antigenic landscape, this research contributes to controlling ASFV's devastating impacts, safeguarding the swine industry, and ensuring food security.
非洲猪瘟病毒(ASFV)是一种极具传染性且往往致命的病原体,对全球养猪业构成重大威胁。H171R蛋白是ASFV的一种结构成分,在病毒组装、宿主细胞进入以及宿主免疫反应调节中发挥关键作用。本研究旨在全面表征H171R蛋白上的线性B细胞表位,以促进针对ASFV的诊断工具和亚单位疫苗的开发。采用了生物信息学分析和实验技术相结合的方法。表达并纯化了重组H171R蛋白,并通过免疫和杂交瘤技术产生了特异性单克隆抗体。使用重叠肽片段和丙氨酸扫描诱变进行系统的表位定位,揭示了四个最小线性表位:HPLLPYQQSSDEQP、SDEQPMMPYQQPPG、PYEQIYHKKHASQQ和LNDYYQHILALGDED。鉴定出的表位具有很强的免疫原性,这通过它们与ASFV阳性猪血清的反应性得到证明。通过突变分析确定了每个表位内的关键氨基酸残基。H171R蛋白的结构建模和可视化提供了表位区域的空间分布和可及性的见解。这些发现有助于更好地理解H171R蛋白的抗原特性,并为开发针对非洲猪瘟的有效诊断方法和亚单位疫苗奠定基础。
非洲猪瘟病毒(ASFV)对全球养猪业构成严重威胁。本研究表征了关键的ASFV H171R蛋白上的线性B细胞表位,促进了改进诊断方法和亚单位疫苗的开发。鉴定出四个免疫原性表位,为设计敏感诊断方法和潜在亚单位疫苗候选物提供了有价值的信息。通过加深对H171R抗原格局的理解,本研究有助于控制ASFV的破坏性影响,保障养猪业并确保食品安全。