Effects of Salmonella enterica serovars Typhimurium (ST) and Choleraesuis (SC) on chemokine and cytokine expression in swine ileum and jejunal epithelial cells
Effects of Salmonella enterica serovars Typhimurium (ST) and Choleraesuis (SC) on chemokine and cytokine expression in swine ileum and jejunal epithelial cells
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DOI：
10.1016/j.vetimm.2006.01.002
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发表时间：
2006-06-15
影响因子：
1.8
通讯作者：
Minton, J. E.
中科院分区：
文献类型：
--
作者：
Skjolaas, K. A.;Burkey, T. E.;Minton, J. E.
胃肠道上皮代表了对潜在侵入性肠道病原体的屏障，在先天免疫监视中保持作用，并且是响应细菌入侵的趋化因子和细胞因子趋化介质的来源。在目前的研究中。我们评价了已知调节巨噬细胞（巨噬细胞迁移抑制因子MIF）、中性粒细胞（IL 8）、树突细胞（CCL 20）和上皮重塑（骨桥蛋白; OPN）运动的细胞因子和趋化因子介体，这些细胞因子和趋化因子介体响应于侵袭性猪肠道病原体沙门氏菌（ST）或猪霍乱（SC）。对于体内实验，断奶猪作为未感染对照（0 h）或经口给予3 x 10（9）CFU ST。在8、24时处死猪。48和144接种后，从近端（PI）和远端（DI）回肠的限定节段提取总RNA。ST对MIF和OPN的相对表达无影响。PI中IL 8表达在24和144 μ l时在数值上增加（相互作用项P = 0.17），这些增加解释了PI中相对于DI的更大表达（P < 0.05）。ST后24 h，CCL 20相对表达量增加（P < 0.05）。接下来，我们评估了MIE IL 8、CCL 20的时间过程。体外培养猪空肠上皮细胞（IPEC-J2），观察脂多糖（LPS）、ST或SC对OPN mRNA表达的影响。使细胞在渗透膜上生长至汇合，并用LPS（10 ng/mL）、ST或SC（10（8）/孔）顶部处理。1小时后，洗涤细胞以除去LPS或细胞外细菌，并加入含有庆大霉素的培养基以杀死剩余的细胞外细菌。在处理后1.5、3和6小时收集培养基和RNA。MIF mRNA不受LPS或细菌处理的影响。类似地，IL 8表达不受LPS影响，但在暴露后1.5和3 μ l，相对于对照，ST和SC增加了IL 8表达（对于所有比较，P < 0.05）。与对照组相比，SC处理在3 h时增加了CCL 20 mRNA（P < 0.05），而ST在1.5、3和6 h时增加了CCL 20，在6 h时表达最大（所有比较P < 0.05）。ST和SC增加极化IL 8分泌。我们的数据表明，侵入性细菌病原体在猪胃肠道触发选定的细胞因子和趋化因子介质的上调，但沙门氏菌血清型引起不同的激活模式在体外。(c)2006 Elsevier B. V.保留所有权利。
The gastrointestinal epithelium represents a barrier to potentially invasive enteric pathogens, maintains a role in innate immune surveillance, and is a source of both chemokine and cytokine chemotactic mediators in response to bacterial invasion. In the current study. we evaluated cytokine and chemokine mediators known to regulate movement of macrophages (macrophage migration inhibitory factor MIF), neutrophils (IL8), dendritic cells (CCL20), and epithelial remodeling (osteopontin; OPN) in response to invasive swine enteropathogens Salmonella enterica serovar Typhimurium (ST) or Choleraesuis (SC). For the in vivo experiment, weaned pigs served as uninfected controls (0 h) or were given 3 x 10(9) CFU ST orally. Pigs were sacrificed at 8, 24. 48, and 144 It after inoculation and total RNA was extracted from defined segments of proximal (PI) and distal (DI) ileum. Relative expression of MIF and OPN were not affected by ST. IL8 expression was increased numerically (P = 0.17 for the interaction term) at 24 and 144 It in the PI and these increases accounted for greater expression in the PI relative to the DI (P < 0.05). Relative expression of CCL20 was increased at 24 h after ST (P < 0.05). Next, we evaluated the time course of MIE IL8, CCL20. and OPN mRNA expression induced by application of lipopolysaccharide (LPS), ST or SC in vitro using pig jejunal epithelial cells (IPEC-J2). Cells were grown to confluency on permeable membranes, and treated apically with LPS (10 ng/mL), ST or SC (10(8)/well). After 1 h, cells were washed to remove LPS or extracellular bacteria, and media containing gentamicin was added to kill remaining extracellular bacteria. Media and RNA were collected at 1.5, 3, and 6 It after treatment. MIF mRNA was not affected by LPS or bacterial treatment. Similarly, IL8 expression was not affected by LPS, but was increased by ST and SC relative to controls at 1.5 and 3 It post exposure (P < 0.05 for all comparisons). Treatment with SC increased CCL20 mRNA relative to controls at 3 h (P < 0.05), while ST increased CCL20 at 1.5, 3, and 6 h with maximal expression at 6 h (P < 0.05 for all comparisons). ST and SC increased polarized IL8 secretion. Our data demonstrate that invasive bacterial pathogens in the pig gastrointestinal tract trigger upregulation of selected cytokine and chemokine mediators, but serovars of Salmonella elicited differing patterns of activation in vitro. (c) 2006 Elsevier B.V. All rights reserved.