Quintana-Hayashi Macarena P, Mahu Maxime, De Pauw Nele, Boyen Filip, Pasmans Frank, Martel An, Premaratne Pushpa, Fernandez Harvey R, Teymournejad Omid, Vande Maele Lien, Haesebrouck Freddy, Lindén Sara K
Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Infect Immun. 2015 Apr;83(4):1610-9. doi: 10.1128/IAI.03073-14. Epub 2015 Feb 2.
Brachyspira hyodysenteriae colonizes the pig colon, resulting in mucohemorrhagic diarrhea and growth retardation. Fecal mucus is a characteristic feature of swine dysentery; therefore, we investigated how the mucin environment changes in the colon during infection with B. hyodysenteriae and how these changes affect this bacterium's interaction with mucins. We isolated and characterized mucins, the main component of mucus, from the colon of experimentally inoculated and control pigs and investigated B. hyodysenteriae binding to these mucins. Fluorescence microscopy revealed a massive mucus induction and disorganized mucus structure in the colon of pigs with swine dysentery. Quantitative PCR (qPCR) and antibody detection demonstrated that the mucus composition of pigs with swine dysentery was characterized by de novo expression of MUC5AC and increased expression of MUC2 in the colon. Mucins from the colon of inoculated and control pigs were isolated by two steps of isopycnic density gradient centrifugation. The mucin densities of control and inoculated pigs were similar, whereas the mucin quantity was 5-fold higher during infection. The level of B. hyodysenteriae binding to mucins differed between pigs, and there was increased binding to soluble mucins isolated from pigs with swine dysentery. The ability of B. hyodysenteriae to bind, measured in relation to the total mucin contents of mucus in sick versus healthy pigs, increased 7-fold during infection. Together, the results indicate that B. hyodysenteriae binds to carbohydrate structures on the mucins as these differ between individuals. Furthermore, B. hyodysenteriae infection induces changes to the mucus niche which substantially increase the amount of B. hyodysenteriae binding sites in the mucus.
猪痢疾短螺旋体定殖于猪的结肠,导致黏液出血性腹泻和生长发育迟缓。粪便黏液是猪痢疾的一个特征性表现;因此,我们研究了感染猪痢疾短螺旋体期间结肠中黏蛋白环境如何变化,以及这些变化如何影响该细菌与黏蛋白的相互作用。我们从实验接种猪和对照猪的结肠中分离并鉴定了黏液的主要成分——黏蛋白,并研究了猪痢疾短螺旋体与这些黏蛋白的结合情况。荧光显微镜检查显示,患有猪痢疾的猪结肠中出现大量黏液诱导和黏液结构紊乱。定量聚合酶链反应(qPCR)和抗体检测表明,患有猪痢疾的猪的黏液组成特征是结肠中MUC5AC从头表达以及MUC2表达增加。通过两步等密度梯度离心法从接种猪和对照猪的结肠中分离出黏蛋白。对照猪和接种猪的黏蛋白密度相似,而感染期间黏蛋白量高出5倍。猪痢疾短螺旋体与黏蛋白的结合水平在不同猪之间存在差异,并且与从患有猪痢疾的猪中分离出的可溶性黏蛋白的结合增加。相对于患病猪与健康猪黏液中总黏蛋白含量来衡量,猪痢疾短螺旋体的结合能力在感染期间增加了7倍。总之,结果表明猪痢疾短螺旋体与黏蛋白上的碳水化合物结构结合,因为这些结构在个体之间存在差异。此外,猪痢疾短螺旋体感染会引起黏液生态位的变化,从而大幅增加黏液中猪痢疾短螺旋体结合位点的数量。