Smith H E, Damman M, van der Velde J, Wagenaar F, Wisselink H J, Stockhofe-Zurwieden N, Smits M A
Department of Bacteriology, Pathobiology and Epidemiology, DLO-Institute for Animal Science and Health, 8200 AB Lelystad, The Netherlands.
Infect Immun. 1999 Apr;67(4):1750-6. doi: 10.1128/IAI.67.4.1750-1756.1999.
To study the role of the capsule of Streptococcus suis serotype 2 in virulence, we generated two isogenic mutants disturbed in capsule production. For that purpose, we first cloned and characterized a major part of the capsular polysaccharide biosynthesis (cps) locus of S. suis serotype 2. Based on the established sequence, 14 open reading frames (ORFs), designated Orf2Z, Orf2Y, Orf2X, and Cps2A to Cps2K, were identified. Twelve ORFs belonged to a single transcriptional unit. The gene products of 11 of these ORFs showed similarity to proteins involved in polysaccharide biosynthesis of other gram-positive microorganisms. Nonencapsulated isogenic mutants were generated in the cps2B and cps2EF genes by insertional mutagenesis. In contrast to the wild-type S. suis serotype 2 strain, the nonencapsulated strains were highly sensitive to ingestion by porcine alveolar lung macrophages in vitro. More importantly, the nonencapsulated mutant strains were completely avirulent in young germfree pigs after intranasal inoculation. These observations indicate that the capsule of S. suis serotype 2 plays an essential role in the pathogenesis of S. suis serotype 2 infections.
为研究猪链球菌2型荚膜在毒力中的作用,我们构建了两个荚膜产生受干扰的同基因突变体。为此,我们首先克隆并鉴定了猪链球菌2型荚膜多糖生物合成(cps)基因座的主要部分。基于已确定的序列,鉴定出14个开放阅读框(ORF),分别命名为Orf2Z、Orf2Y、Orf2X以及Cps2A至Cps2K。其中12个ORF属于单个转录单元。这些ORF中的11个基因产物与其他革兰氏阳性微生物多糖生物合成相关蛋白具有相似性。通过插入诱变在cps2B和cps2EF基因中产生了无荚膜的同基因突变体。与野生型猪链球菌2型菌株相比,无荚膜菌株在体外对猪肺泡巨噬细胞的吞噬高度敏感。更重要的是,无荚膜突变菌株经鼻内接种后,在幼龄无菌猪中完全无毒力。这些观察结果表明,猪链球菌2型荚膜在猪链球菌2型感染的发病机制中起重要作用。