Charland Nathalie, Harel Josée, Kobisch Marylène, Lacasse Serge, Gottschalk Marcelo
Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de médecine vétérinaire, Université de Montréal, CP 5000, St-Hyacinthe, Québec, Canada J2S 7C6.
Centre National d'Études Vétérinaires et alimentaires, Unité Mycoplasmologie Bactériologie, BP 53, Zoopôle, 22440 Ploufragan, France.
Microbiology (Reading). 1998 Feb;144 ( Pt 2):325-332. doi: 10.1099/00221287-144-2-325.
Streptococcus suis serotype 2 is responsible for a wide variety of porcine infections. In addition, it is considered a zoonotic agent. Knowledge about the virulence factors for this bacterium is limited but its polysaccharide capsule is thought to be one of the most important. Transposon mutagenesis with the self-conjugative transposon Tn916 was used to obtain acapsular mutants from the virulent S. suis type 2 reference strain S735. Clones were screened by colony-dot ELISA with a monoclonal antibody specific for a type 2 capsular epitope and clones that failed to react with the antibody were characterized. Two mutants, 2A and 79, having one and two Tn916 insertions respectively, were chosen for further characterization. Absence of capsule was confirmed by coagglutination, capillary precipitation and capsular reaction tests and by transmission electron microscopy. Absence of capsular polysaccharides correlated with increased hydrophobicity and phagocytosis by both murine macrophages and porcine monocytes compared to the wild-type strain. Furthermore, both mutants were shown to be avirulent in murine and pig models of infection. Finally, mutant 2A was readily eliminated from circulation in mice compared to the wild-type strain, which persisted more than 48 h in blood. Thus, isogenic mutants defective in capsule production demonstrate the importance of capsular polysaccharides as a virulence factor for S. suis type 2.
猪链球菌2型可引发多种猪感染。此外,它被认为是一种人畜共患病原体。关于这种细菌毒力因子的知识有限,但其多糖荚膜被认为是最重要的毒力因子之一。利用自共轭转座子Tn916进行转座子诱变,从强毒猪链球菌2型参考菌株S735获得无荚膜突变体。通过用针对2型荚膜表位的单克隆抗体进行菌落斑点ELISA筛选克隆,并对不与该抗体反应的克隆进行特性分析。选择分别有一个和两个Tn916插入的两个突变体2A和79进行进一步特性分析。通过协同凝集、毛细管沉淀和荚膜反应试验以及透射电子显微镜证实无荚膜。与野生型菌株相比,无荚膜多糖与小鼠巨噬细胞和猪单核细胞的疏水性增加及吞噬作用增强相关。此外,在小鼠和猪感染模型中,两个突变体均显示无毒性。最后,与野生型菌株相比,突变体2A在小鼠体内循环中很容易被清除,野生型菌株在血液中持续存在超过48小时。因此,荚膜产生缺陷的同基因突变体证明了荚膜多糖作为猪链球菌2型毒力因子的重要性。