Department of Infectious Diseases and Preventive Medicine, Veterinary Research Institute, Brno, Czechia.
Department of Pharmacology and Toxicology, Veterinary Research Institute, Brno, Czechia.
Front Immunol. 2021 Apr 22;12:635097. doi: 10.3389/fimmu.2021.635097. eCollection 2021.
In Glässer's disease outbreaks, has to overcome the non-specific immune system in the lower respiratory tract, the alveolar macrophages. Here we showed that porcine alveolar macrophages (PAMs) were able to recognize and phagocyte with strain-to-strain variability despite the presence of the capsule in virulent (serovar 1, 5, 12) as well in avirulent strains (serovar 6 and 9). The capsule, outer membrane proteins, virulence-associated autotransporters, cytolethal distending toxins and many other proteins have been identified as virulence factors of this bacterium. Therefore, we immunized pigs with the crude capsular extract (cCE) from the virulent CAPM 6475 strain (serovar 5) and evaluated the role of the anti-cCE/post-vaccinal IgG in the immune response of PAMs to infection with various strains. We demonstrated the specific binding of the antibodies to the cCE by Western-blotting assay and immunoprecipitation as well as the specific binding to the strain CAPM 6475 in transmission electron microscopy. In the cCE, we identified several virulence-associated proteins that were immunoreactive with IgG isolated from sera of immunized pigs. Opsonization of strains by post-vaccinal IgG led to enhanced phagocytosis of by PAMs at the first two hours of infection. Moreover, opsonization increased the oxidative burst and expression/production of both pro- and anti-inflammatory cytokines. The neutralizing effects of these antibodies on the antioxidant mechanisms of may lead to attenuation of its virulence and pathogenicity . Together with opsonization of bacteria by these antibodies, the host may eliminate in the infection site more efficiently. Based on these results, the crude capsular extract is a vaccine candidate with immunogenic properties.
在格拉泽氏病暴发中,必须克服下呼吸道的非特异性免疫系统,即肺泡巨噬细胞。在这里,我们表明,尽管有荚膜的存在,猪肺泡巨噬细胞 (PAMs) 能够识别和吞噬具有毒力的菌株(血清型 1、5、12)以及无毒性菌株(血清型 6 和 9)。荚膜、外膜蛋白、与毒力相关的自动转运蛋白、细胞致死性扩张毒素和许多其他蛋白质已被确定为该细菌的毒力因子。因此,我们用从强毒株(血清型 5)CAPM 6475 菌株中提取的粗荚膜提取物 (cCE) 对猪进行免疫接种,并评估抗 cCE/疫苗后 IgG 在 PAMs 对各种 菌株感染的免疫反应中的作用。我们通过 Western-blotting 分析和免疫沉淀证明了抗体与 cCE 的特异性结合,以及在透射电子显微镜下与菌株 CAPM 6475 的特异性结合。在 cCE 中,我们鉴定了几种与毒力相关的蛋白质,它们与免疫接种猪血清中分离的 IgG 发生免疫反应。疫苗后 IgG 对 菌株的调理作用导致感染后前两小时 PAMs 对 的吞噬作用增强。此外,调理作用增加了氧化爆发和促炎和抗炎细胞因子的表达/产生。这些抗体对 的抗氧化机制的中和作用可能导致其毒力和致病性减弱。结合这些抗体对细菌的调理作用,宿主可能更有效地在感染部位清除 。基于这些结果,粗荚膜提取物是一种具有免疫原性的候选疫苗。