Key Laboratory of Animal Disease Diagnostic & Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
Proteome Sci. 2011 Jun 26;9(1):32. doi: 10.1186/1477-5956-9-32.
Actinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide. Identification and characterization of novel antigenic APP vaccine candidates are underway. In the present study, we use an immunoproteomic approach to identify APP protein antigens that may elicit an immune response in serotype 1 naturally infected swine and serotype 1 virulent strain S259-immunized rabbits.
Proteins from total cell lysates of serotype 1 APP were separated by two-dimensional electrophoresis (2DE). Western blot analysis revealed 21 immunoreactive protein spots separated in the pH 4-7 range and 4 spots in the pH 7-11 range with the convalescent sera from swine; we found 5 immunoreactive protein spots that separated in the pH 4-7 range and 2 in the pH 7-11 range with hyperimmune sera from S259-immunized rabbits. The proteins included the known antigens ApxIIA, protective surface antigen D15, outer membrane proteins P5, subunit NqrA. The remaining antigens are being reported as immunoreactive proteins in APP for the first time, to our knowledge.
We identified a total of 42 immunoreactive proteins of the APP serotype 1 virulent strain S259 which represented 32 different proteins, including some novel immunoreactive factors which could be researched as vaccine candidates.
胸膜肺炎放线杆菌（APP）是世界范围内最重要的猪病原体之一。目前正在鉴定和表征新型抗原 APP 疫苗候选物。在本研究中，我们使用免疫蛋白质组学方法来鉴定可能在血清型 1 自然感染猪和血清型 1 强毒力菌株 S259 免疫兔中引起免疫反应的 APP 蛋白抗原。
血清型 1 APP 总细胞裂解物的蛋白质通过二维电泳（2DE）分离。Western blot 分析显示，与恢复期猪血清相比，pH 值为 4-7 的 21 个免疫反应性蛋白斑点和 pH 值为 7-11 的 4 个蛋白斑点分离；与 S259 免疫兔的高免血清相比，pH 值为 4-7 的 5 个免疫反应性蛋白斑点和 pH 值为 7-11 的 2 个蛋白斑点分离。这些蛋白包括已知的抗原 ApxIIA、保护性表面抗原 D15、外膜蛋白 P5、亚基 NqrA。据我们所知，其余抗原是首次被报道为 APP 中的免疫反应性蛋白。
我们共鉴定了血清型 1 强毒力菌株 S259 的 42 种免疫反应性蛋白，代表了 32 种不同的蛋白，其中包括一些新的免疫反应性因子，可作为疫苗候选物进行研究。
