Department of Veterinary and Animal Sciences, Stigbøjlen 4, 1870 Frb. C., 1-20, Building: 301, University of Copenhagen, Copenhagen, Denmark.
Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124, Braunschweig, Germany.
Vet Microbiol. 2019 Aug;235:188-194. doi: 10.1016/j.vetmic.2019.07.001. Epub 2019 Jul 3.
Outer membrane vesicles (OMVs) are produced and secreted virtually by every known Gram-negative bacterium. Despite their non-live nature, they share antigenic characteristics with the bacteria they originate from. This, together with their relative ease of purification, casts the OMVs as a very promising and flexible tool in both human and veterinary vaccinology. The aim of the current work was to get an insight into the antigenic pattern of OMVs from the pig pathogen Actinobacillus pleuropneumoniae in the context of vaccine development. Accordingly, we designed a protocol combining 2D Western Blotting and mass spectrometric identification to robustly characterize the antigenic protein pattern of the vesicles. Our analysis revealed that A. pleuropneumoniae OMVs carry several immunoreactive virulence factors. Some of these proteins, LpoA, OsmY and MIDG2331_02184, have never previously been documented as antigenic in A. pleuropneumoniae or other pathogenic bacteria. Additionally, we showed that despite their relative abundance, proteins such as FrpB and DegQ do not contribute to the antigenic profile of A. pleuropneumoniae OMVs.
外膜囊泡（OMVs）几乎可以由所有已知的革兰氏阴性菌产生和分泌。尽管它们是非活体的，但它们与它们起源的细菌具有共同的抗原特征。这一点，再加上它们相对容易纯化，使得 OMV 成为人类和兽医疫苗学中一种非常有前途和灵活的工具。目前这项工作的目的是深入了解猪病原体胸膜肺炎放线杆菌 OMV 的抗原模式，以促进疫苗的开发。因此，我们设计了一种结合 2D 印迹和质谱鉴定的方案，以稳健地表征囊泡的抗原蛋白模式。我们的分析表明，胸膜肺炎放线杆菌 OMV 携带多种免疫反应性毒力因子。其中一些蛋白，LpoA、OsmY 和 MIDG2331_02184，以前从未在胸膜肺炎放线杆菌或其他病原菌中被记录为抗原。此外，我们还表明，尽管 FrpB 和 DegQ 等蛋白相对丰富，但它们并不构成胸膜肺炎放线杆菌 OMV 抗原谱的一部分。
