The Levels of Brachyspira hyodysenteriae Binding to Porcine Colonic Mucins Differ between Individuals, and Binding Is Increased to Mucins from Infected Pigs with De Novo MUC5AC Synthesis
The Levels of Brachyspira hyodysenteriae Binding to Porcine Colonic Mucins Differ between Individuals, and Binding Is Increased to Mucins from Infected Pigs with De Novo MUC5AC Synthesis
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DOI：
10.1128/iai.03073-14
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发表时间：
2015-04-01
影响因子：
3.1
通讯作者：
Linden, Sara K.
中科院分区：
文献类型：
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作者：
Quintana-Hayashi, Macarena P.;Mahu, Maxime;Linden, Sara K.
猪痢疾杆菌定植于猪的结肠，导致粘液出血性腹泻和生长迟缓。粪便粘液是猪痢疾的一个特征；因此，我们调查了感染猪痢疾杆菌期间结肠中粘蛋白环境的变化，以及这些变化如何影响这种细菌与粘蛋白的相互作用。我们从实验接种猪和对照猪的结肠中分离和鉴定了粘液的主要成分粘蛋白，并研究了猪痢疾杆菌与这些粘蛋白的结合。荧光显微镜下可见猪痢疾猪结肠内可见大量粘液诱导和粘液结构紊乱。定量聚合酶链式反应(QPCR)和抗体检测表明，猪痢疾的粘液成分以MUC5AC的从头表达和MUC2在结肠的表达增加为特征。用两步等密度梯度离心法从接种猪和对照猪的结肠中分离粘蛋白。对照猪和接种猪的粘蛋白密度相似，而感染过程中粘蛋白的数量增加了5倍。猪痢疾杆菌与粘蛋白的结合水平在不同猪之间不同，与从猪痢疾中分离的可溶性粘蛋白的结合增加。猪痢疾杆菌的结合能力，与患病猪与健康猪粘液的总粘蛋白含量相比，在感染期间增加了7倍。总之，结果表明猪痢疾杆菌与粘蛋白上的碳水化合物结构结合，因为这些结构在个体之间是不同的。此外，猪痢疾杆菌感染会引起粘液生态位的变化，这大大增加了粘液中猪痢疾杆菌结合位点的数量。
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.