Response of Porcine Intestinal In Vitro Organ Culture Tissues following Exposure to Lactobacillus plantarum JC1 and Salmonella enterica Serovar Typhimurium SL1344
Response of Porcine Intestinal In Vitro Organ Culture Tissues following Exposure to Lactobacillus plantarum JC1 and Salmonella enterica Serovar Typhimurium SL1344
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DOI：
10.1128/aem.03115-09
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发表时间：
2010-10-01
影响因子：
4.4
通讯作者：
La Ragione, R. M.
中科院分区：
文献类型：
--
作者：
Collins, J. W.;Coldham, N. G.;La Ragione, R. M.
开发新的干预策略来控制由沙门氏菌等细菌引起的人畜共患病。在家畜中需要适当的实验模型来评估其适用性。这里，开发了一种利用细胞冠（CC）技术（CCIVOC）（Scaffdex）的新型猪肠道体外器官培养（IVOC）模型。采用 CCIVOC 模型研究鼠伤寒沙门氏菌菌株 SL1344 在接触植物乳杆菌菌株后与猪肠道组织的关联特征。经过适当的处理和染色后，通过光学显微镜和电子显微镜（EM）检查细菌与宿主细胞的关联，同时使用定量无标记蛋白质组学研究猪空肠组织蛋白质组的变化。将猪肠粘膜组织暴露于植物乳杆菌 JC1 并不会减少与组织相关的鼠伤寒沙门氏菌数量，但会显着 (P < 0.005) 减少肠道 IVOC 组织区域的百分比，从而给出酸性粘蛋白阳性染色结果。相反，IVOC 组织的杯状细胞内存在的中性电荷粘蛋白的数量显着增加（P < 0.05）。此外，在用植物乳杆菌接种空肠IVOC组织后，微管蛋白-α以高水平表达。尽管植物乳杆菌 JC1 并未减少鼠伤寒沙门氏菌菌株 SL1344 与空肠 IVOC 组织的关联，但对酸性粘蛋白分泌增加、宿主细胞骨架重排和参与猪免疫反应的蛋白质的检测表明，植物乳杆菌菌株可能有助于保护猪免受鼠伤寒沙门氏菌或其他病原体的感染。
The development of novel intervention strategies for the control of zoonoses caused by bacteria such as Salmonella spp. in livestock requires appropriate experimental models to assess their suitability. Here, a novel porcine intestinal in vitro organ culture (IVOC) model utilizing cell crown (CC) technology (CCIVOC) (Scaffdex) was developed. The CCIVOC model was employed to investigate the characteristics of association of S. enterica serovar Typhimurium strain SL1344 with porcine intestinal tissue following exposure to a Lactobacillus plantarum strain. The association of bacteria to host cells was examined by light microscopy and electron microscopy (EM) after appropriate treatments and staining, while changes in the proteome of porcine jejunal tissues were investigated using quantitative label-free proteomics. Exposure of porcine intestinal mucosal tissues to L. plantarum JC1 did not reduce the numbers of S. Typhimurium bacteria associating to the tissues but was associated with significant (P < 0.005) reductions in the percentages of areas of intestinal IVOC tissues giving positive staining results for acidic mucins. Conversely, the quantity of neutrally charged mucins present within the goblet cells of the IVOC tissues increased significantly (P < 0.05). In addition, tubulin-alpha was expressed at high levels following inoculation of jejunal IVOC tissues with L. plantarum. Although L. plantarum JC1 did not reduce the association of S. Typhimurium strain SL1344 to the jejunal IVOC tissues, detection of increased acidic mucin secretion, host cytoskeletal rearrangements, and proteins involved in the porcine immune response demonstrated that this strain of L. plantarum may contribute to protecting the pig from infections by S. Typhimurium or other pathogens.