Inhibitory effects of Lactobacillus casei subsp. rhamnosus on Salmonella lipopolysaccharide-induced inflammation and epithelial barrier dysfunction in a co-culture model using Caco-2/peripheral blood mononuclear cells

Fang, Hsu Wei; Fang, Shiuh-Bin; Chiau, Jen-Shiu Chiang; Yeung, Chun‐Yan; Chan, Wai Tao; Jiang, Chuen-Bin; Cheng, Ming Fang; Lee, Hung Chang

doi:10.1099/jmm.0.009662-0

Cited by 54 publications

(47 citation statements)

References 31 publications

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“…At this time, the co-cultures were inflamed with 10 ng/ml of IL-1b added into the apical compartment. TEER values decreased significantly compared to a non-inflamed cell line-based co-culture, which agrees with previous models based on primary cells (Fang et al, 2010;Leonard et al, 2010). To evaluate the reversible effect of inflammation on barrier properties, IL-1b was removed from the co-cultures on day 23.…”

Section: Replacement Of Primary Cells By Cell Linessupporting

confidence: 86%

A 3D co-culture of three human cell lines to model the inflamed intestinal mucosa for safety testing of nanomaterials

et al. 2015

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CitationA 3D co-culture of three human cell lines to model the inflamed intestinal mucosa for safety testing of nanomaterials. AbstractOral exposure to nanomaterials is a current concern, asking for innovative biological test systems to assess their safety, especially also in conditions of inflammatory disorders. Aim of this study was to develop a 3D intestinal model, consisting of Caco-2 cells and two human immune cell lines, suitable to assess nanomaterial toxicity, in either healthy or diseased conditions. Human macrophages (THP-1) and human dendritic cells (MUTZ-3) were embedded in a collagen scaffold and seeded on the apical side of transwell inserts. Caco-2 cells were seeded on top of this layer, forming a 3D model of the intestinal mucosa. Toxicity of engineered nanoparticles (NM101 TiO 2 , NM300 Ag, Au) was evaluated in non-inflamed and inflamed co-cultures, and also compared to non-inflamed Caco-2 monocultures. Inflammation was elicited by IL-1b, and interactions with engineered NPs were addressed by different endpoints. The 3D co-culture showed well preserved ultrastructure and significant barrier properties. Ag NPs were found to be more toxic than TiO 2 or Au NPs. But once inflamed with IL-1b, the co-cultures released higher amounts of IL-8 compared to Caco-2 monocultures. However, the cytotoxicity of Ag NPs was higher in Caco-2 monocultures than in 3D co-cultures. The naturally higher IL-8 production in the co-cultures was enhanced even further by the Ag NPs. This study shows that it is possible to mimic inflamed conditions in a 3D co-culture model of the intestinal mucosa. The fact that it is based on three easily available human cell lines makes this model valuable to study the safety of nanomaterials in the context of inflammation.

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Section: Replacement Of Primary Cells By Cell Linessupporting

confidence: 86%

A 3D co-culture of three human cell lines to model the inflamed intestinal mucosa for safety testing of nanomaterials

et al. 2015

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“…In this study, dietary zinc supplementation increased the number of bacteria in the Lactobacillus group and the total bacteria. Lactobacillus bacteria are known for their potential to prevent the colonization of pathogens and ameliorate barrier dysfunction by competing for epithelial binding sites as well as through the production of antimicrobial and anti-inflammatory factors (Jones and Versalovic, 2009;Fang, 2010). These results indicated that zinc supplementation in the diet may help maintaining the stability of the intestinal microflora, increasing the number of beneficial bacteria and reducing the chance of S. typhimurium colonization in the cecum, which is in line with the improved intestinal mucosal morphological structure and intestinal function (Sommer and Backhed, 2013).…”

Section: Discussionmentioning

confidence: 99%

Effect of zinc on growth performance, gut morphometry, and cecal microbial community in broilers challenged with Salmonella enterica serovar typhimurium

et al. 2014

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To evaluate the effects of supplemental zinc on growth performance, gut morphometry, and the cecal microbial community in broilers challenged with Salmonella typhimurium, 180, 1-day-old male Cobb 500 broiler chicks were randomly assigned to 3 treatments with ten replicates for a 42 day experiment. The 3 treatments were: unchallenged, S. typhimurium-challenged, and S. typhimurium-challenged with 120 mg/kg of zinc supplementation in the diet. Salmonella infection caused a reduction in body-weight gain and feed intake, disrupted the intestinal structure by decreasing the villus-height/crypt-depth ratio of the ileum and increasing the apoptotic index of ileal epithelial cells. Moreover, the cecal microbial community was altered by Salmonella infection, as demonstrated by a reduced number of Lactobacillus and total bacteria. Dietary zinc supplementation improved growth performance by increasing the body-weight gain and feed intake in the challenged broilers. In addition, zinc repaired intestinal injury by reducing the apoptotic index of ileal epithelial cells, enhancing villus height and the villus-height/crypt-depth ratio of the ileum, and the proliferation index of ileal epithelial cells. Finally, zinc regulated the cecal microbial community by increasing the number of total bacteria and beneficial Lactobacillus bacteria, and reducing the number of Salmonella. The results indicated that dietary zinc supplementation improved growth performance, intestinal morphology, and intestinal microbiota in S. typhimurium-challenged broilers.

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“…Moreover, effects of commensal gut bacteria on the invasion capacity of Salmonella have been observed. Lactobacilli and bifidobacteria, often used as probiotics, can attenuate Salmonella invasion by strengthening the epithelial barrier functions [21], [22], [23], [24], [25], [26]. The gut microbiota might also stimulate immune defenses of the host against pathogens [16], [26], [27].…”

Section: Introductionmentioning

confidence: 99%

Salmonella Adhesion, Invasion and Cellular Immune Responses Are Differentially Affected by Iron Concentrations in a Combined In Vitro Gut Fermentation-Cell Model

et al. 2014

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In regions with a high infectious disease burden, concerns have been raised about the safety of iron supplementation because higher iron concentrations in the gut lumen may increase risk of enteropathogen infection. The aim of this study was to investigate interactions of the enteropathogen Salmonella enterica ssp. enterica Typhimurium with intestinal cells under different iron concentrations encountered in the gut lumen during iron deficiency and supplementation using an in vitro colonic fermentation system inoculated with immobilized child gut microbiota combined with Caco-2/HT29-MTX co-culture monolayers. Colonic fermentation effluents obtained during normal, low (chelation by 2,2'-dipyridyl) and high iron (26.5 mg iron/L) fermentation conditions containing Salmonella or pure Salmonella cultures with similar iron conditions were applied to cellular monolayers. Salmonella adhesion and invasion capacity, cellular integrity and immune response were assessed. Under high iron conditions in pure culture, Salmonella adhesion was 8-fold increased compared to normal iron conditions while invasion was not affected leading to decreased invasion efficiency (−86%). Moreover, cellular cytokines IL-1β, IL-6, IL-8 and TNF-α secretion as well as NF-κB activation in THP-1 cells were attenuated under high iron conditions. Low iron conditions in pure culture increased Salmonella invasion correlating with an increase in IL-8 release. In fermentation effluents, Salmonella adhesion was 12-fold and invasion was 428-fold reduced compared to pure culture. Salmonella in high iron fermentation effluents had decreased invasion efficiency (−77.1%) and cellular TNF-α release compared to normal iron effluent. The presence of commensal microbiota and bacterial metabolites in fermentation effluents reduced adhesion and invasion of Salmonella compared to pure culture highlighting the importance of the gut microbiota as a barrier during pathogen invasion. High iron concentrations as encountered in the gut lumen during iron supplementation attenuated Salmonella invasion efficiency and cellular immune response suggesting that high iron concentrations alone may not lead to an increased Salmonella invasion.

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Inhibitory effects of Lactobacillus casei subsp. rhamnosus on Salmonella lipopolysaccharide-induced inflammation and epithelial barrier dysfunction in a co-culture model using Caco-2/peripheral blood mononuclear cells

Cited by 54 publications

References 31 publications

A 3D co-culture of three human cell lines to model the inflamed intestinal mucosa for safety testing of nanomaterials

A 3D co-culture of three human cell lines to model the inflamed intestinal mucosa for safety testing of nanomaterials

Effect of zinc on growth performance, gut morphometry, and cecal microbial community in broilers challenged with Salmonella enterica serovar typhimurium

Salmonella Adhesion, Invasion and Cellular Immune Responses Are Differentially Affected by Iron Concentrations in a Combined In Vitro Gut Fermentation-Cell Model

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