Image_1_Lactobacillus acidophilus Attenuates Salmonella-Induced Stress of Epithelial Cells by Modulating Tight-Junction Genes and Cytokine Responses.PDF

<p>Scope: Salmonellosis is a prevalent food-borne illness that causes diarrhea in over 130 million humans yearly and can lead to death. There is an urgent need to find alternatives to antibiotics as many salmonellae are now multidrug resistant. As such, specific beneficial bacteria and dietary fibers can be an alternative as they may prevent Salmonella Typhimurium (STM) infection and spreading by strengthening intestinal barrier function.</p><p>Methods and Results: We tested whether immune active long-chain inulin-type fructans and/or L. acidophilus W37, L. brevis W63, and L. casei W56 can strengthen barrier integrity of intestinal Caco-2 cells in the presence and absence of a STM. Effects of the ingredients on intestinal barrier function were first evaluated by quantifying trans-epithelial electric resistance (TEER) and regulation of gene expression by microarray. Only L. acidophilus had effects on TEER and modulated a group of 26 genes related to tight-junctions. Inulin-type fructans, L. brevis W63 and L. casei W56 regulated other genes, unrelated to tight-junctions. L. acidophilus also had unique effects on a group of six genes regulating epithelial phenotype toward follicle-associated epithelium. L. acidophilus W37 was therefore selected for a challenge with STM and prevented STM-induced barrier disruption and decreased secretion of IL-8.</p><p>Conclusion:L. acidophilus W37 increases TEER and can protect against STM induced disruption of gut epithelial cells integrity in vitro. Our results suggest that selection of specific bacterial strains for enforcing barrier function may be a promising strategy to reduce or prevent STM infections.</p

Median (IQR) of serum levels of all detected molecules (pg/mL) in Crohn’s disease patients with ‘normal’ (< 200 mg/kg) and increased (> 200 mg/kg) fecal calprotectin levels.

<p>Median (IQR) of serum levels of all detected molecules (pg/mL) in Crohn’s disease patients with ‘normal’ (< 200 mg/kg) and increased (> 200 mg/kg) fecal calprotectin levels.</p

Distribution of serum biomarker levels in patients with normal (blue, < 200 mg/kg) and increased (red, > 200 mg/kg) fecal calprotectin levels, shown in boxplots.

<p>(A) Serum IL-6 levels (pg/mL). (B) Serum IL-17A levels (pg/mL). (C) Serum TNF-β levels (pg/mL). (D) Serum SAA levels (pg/mL). *<i>P</i> < 0.05; **<i>P</i> < 0.01.</p

Data_Sheet_1_Lactobacillus acidophilus Attenuates Salmonella-Induced Stress of Epithelial Cells by Modulating Tight-Junction Genes and Cytokine Responses.XLSX

<p>Scope: Salmonellosis is a prevalent food-borne illness that causes diarrhea in over 130 million humans yearly and can lead to death. There is an urgent need to find alternatives to antibiotics as many salmonellae are now multidrug resistant. As such, specific beneficial bacteria and dietary fibers can be an alternative as they may prevent Salmonella Typhimurium (STM) infection and spreading by strengthening intestinal barrier function.</p><p>Methods and Results: We tested whether immune active long-chain inulin-type fructans and/or L. acidophilus W37, L. brevis W63, and L. casei W56 can strengthen barrier integrity of intestinal Caco-2 cells in the presence and absence of a STM. Effects of the ingredients on intestinal barrier function were first evaluated by quantifying trans-epithelial electric resistance (TEER) and regulation of gene expression by microarray. Only L. acidophilus had effects on TEER and modulated a group of 26 genes related to tight-junctions. Inulin-type fructans, L. brevis W63 and L. casei W56 regulated other genes, unrelated to tight-junctions. L. acidophilus also had unique effects on a group of six genes regulating epithelial phenotype toward follicle-associated epithelium. L. acidophilus W37 was therefore selected for a challenge with STM and prevented STM-induced barrier disruption and decreased secretion of IL-8.</p><p>Conclusion:L. acidophilus W37 increases TEER and can protect against STM induced disruption of gut epithelial cells integrity in vitro. Our results suggest that selection of specific bacterial strains for enforcing barrier function may be a promising strategy to reduce or prevent STM infections.</p

Study population characteristics (<i>n</i> = 39) of Crohn’s disease patients with ‘normal’ (< 200 mg/kg) and increased (> 200 mg/kg) fecal calprotectin levels.

<p>Study population characteristics (<i>n</i> = 39) of Crohn’s disease patients with ‘normal’ (< 200 mg/kg) and increased (> 200 mg/kg) fecal calprotectin levels.</p

Confocal microscopy images of alginate-PLL-PEG-b-PLL microcapsules after the addition of a) dextran of 110 kg/mol and b) dextran of 150 kg/mol.

<p>Confocal microscopy images of alginate-PLL-PEG-b-PLL microcapsules after the addition of a) dextran of 110 kg/mol and b) dextran of 150 kg/mol.</p

Correlations of fecal calprotectin levels (mg/kg) with serum levels of all detected molecules (pg/mL) in patients with Crohn’s disease.

<p>Correlations of fecal calprotectin levels (mg/kg) with serum levels of all detected molecules (pg/mL) in patients with Crohn’s disease.</p

Distribution of serum biomarker levels in patients with normal (blue, < 200 mg/kg) and increased (red, > 200 mg/kg) fecal calprotectin levels, shown in boxplots.

<p>(A) Serum CRP levels (pg/mL). (B) Serum IFN-γ levels (pg/mL). *<i>P</i> < 0.05; **<i>P</i> < 0.01.</p

An representative example of a FACS analysis procedure to evaluate lymphocytes subpopulations.

<p>[A] A forward/sideward (FSC/SSC) scatterplot of all events in which leukocytes were identified. [B] The leukocyte gate was copied to a FSC/SSC scatterplot in which the lymphocytes are identified. [C] These lymphocytes are copied to a SSC/CD3-PerCP scatterplot to distinguish T-lymphocytes (CD3⁺). [D] The lymphocytes are further divided into helper (CD3⁺/CD4⁺) and cytotoxic (CD3^+/CD4⁺; ct) T-cells in a CD3 PerCP/CD4 Alexa 700 scatterplot. [E] A CD3 PerCP/CD25 FITC scatterplot in which CD3⁻ cells were copied to assess the gate for FITC-positivity. [F] This gate for FITC positivity was copied to a CD4 Alexa 700/CD25 FITC scatterplot to identify CD25 positive and negative cells; i.e. effector (CD3⁺/CD4⁺/CD25⁺) and naive helper T-cells (CD3⁺/CD4⁺/CD25⁻). [G] A CD25 FITC/FoxP3 Alexa 647 scatterplot to identify the regulatory T-cell (CD3⁺/CD4⁺/CD25⁺/FoxP3⁺; Treg) positive population.</p

Lymphocyte subpopulations in pregnant rats.

<p>Values of day 10 or 18 of pregnancy are recalculated regarding the median values of non-pregnant animals (which was set at 100%) to express (as median (Q₁–Q₃)) pregnancy-induced changes. [A] The adaptations to pregnancy in percentage of T-lymphocytes, ratio of T-helper cells (Th) and cytotoxic T-cells (Tc), percentages of effector T-cells (Teff), regulatory T-cells (Treg), NK-cells and the ratio of MFI CD161a/NKR-P1b of NK-cells in all three strains during pregnancy. [B] The adaptations to pregnancy of the Th1/Th2-mRNA ratio and ROR-C mRNA expression in all three strains during pregnancy. d10/d18: day 10 or 18 of pregnancy. ∞slope significantly different from zero (Regression-analysis, ANCOVA, p<0.05). *slope significant different between the marked strains (Regression-analysis, ANCOVA, p<0.05).</p