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Rhonda F Souza - One of the best experts on this subject based on the ideXlab platform.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Eunice Odiase, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    Background & Aims In upper airway cells, T helper 2 cytokines that signal through interleukin-4 (IL-4) receptor-α have been shown to stimulate Eotaxin-3 secretion via a nongastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL-4 receptor-α signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by quantitative real-time polymerase chain reaction and Western blotting. IL-4–stimulated Eotaxin-3 secretion was measured by enzyme-linked immunosorbent assay after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester (calcium chelator), 2-aminoethoxydiphenyl borate (inhibitor of endoplasmic reticulum calcium release), verapamil, and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of patients with EoE and controls. Results EoE cells expressed ngH+,K+ATPase messenger RNA and protein. Omeprazole and SCH 28080 decreased IL-4–stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4–stimulated Eotaxin-3 secretion was blocked by ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester, 2-aminoethoxydiphenyl borate, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4–stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a nongastric proton pump that mediates T helper 2 cytokine–stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block T helper 2 cytokine–stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Eunice Odiase, Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    ABSTRACT Background & Aims In upper airway cells, Th2 cytokines that signal through IL-4 receptor alpha (IL4Rα) have been shown to stimulate Eotaxin-3 secretion via a non-gastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL4Rα signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by qPCR and western blotting. IL-4-stimulated Eotaxin-3 secretion was measured by ELISA after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), EGTA-AM (calcium chelator), 2-APB (inhibitor of endoplasmic reticulum calcium release), verapamil and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of EoE patients and controls. Results EoE cells expressed ngH+,K+ATPase mRNA and protein. Omeprazole and SCH 28080 decreased IL-4-stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4-stimulated Eotaxin-3 secretion was blocked by EGTA-AM, 2-APB, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4-stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a non-gastric proton pump that mediates Th2 cytokine-stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block Th2 cytokine-stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • jak stat6 pathway inhibitors block Eotaxin 3 secretion by epithelial cells and fibroblasts from esophageal eosinophilia patients promising agents to improve inflammation and prevent fibrosis in eoe
    2016
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Jason Y Park, Kathleen S Wilson, Rhonda F Souza
    Abstract:

    Background: Although most studies on treatments for eosinophilic esophagitis (EoE) have focused on effects in the epithelium, EoE is a transmural disease. Eosinophils that infiltrate the subepithelial layers of the esophagus lead to fibrosis and the serious complications of EoE, and current therapies have shown minimal effects on this fibrosis. We aimed to elucidate T helper (Th)2 cytokine effects on esophageal fibroblasts and to explore potential fibroblast-targeted therapies for EoE. Methods: We established telomerase-immortalized fibroblasts from human esophageal biopsies. We stimulated these esophageal fibroblasts with Th2 cytokines, and examined effects of omeprazole and inhibitors of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT6) pathway (AS1517499, leflunomide, and ruxolitinib) on STAT6 phosphorylation, STAT6 nuclear translocation, and Eotaxin-3 expression. We also measured the effects of these inhibitors in esophageal epithelial cells stimulated with Th2 cytokines. Results: As in esophageal epithelial cells, Th2 cytokines increased STAT6 phosphorylation, STAT6 nuclear translocation, Eotaxin-3 transcription and protein secretion in esophageal fibroblasts. Unlike in epithelial cells, however, omeprazole did not inhibit cytokine-stimulated Eotaxin-3 expression in fibroblasts. In contrast, JAK-STAT6 pathway inhibitors decreased cytokine-stimulated Eotaxin-3 expression in both fibroblasts and epithelial cells. Conclusions: Omeprazole does not inhibit Th2 cytokine-stimulated Eotaxin-3 expression by esophageal fibroblasts, suggesting that PPIs will have limited impact on subepithelial EoE processes such as fibrosis. JAK-STAT6 pathway inhibitors block Th2 cytokine-stimulated Eotaxin-3 expression both in fibroblasts and in epithelial cells, suggesting a potential role for JAK-STAT inhibitors in treating both epithelial inflammation and subepithelial fibrosis in EoE.

  • proton pump inhibitors decrease Eotaxin 3 expression in the proximal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Objective Besides reducing gastric acid secretion, proton pump inhibitors (PPIs) suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal epithelial cells through acid-independent, anti-inflammatory mechanisms. To explore acid-inhibitory and acid-independent, anti-inflammatory PPI effects in reducing esophageal eosinophilia, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid and bile salt effects on IL-13-stimulated Eotaxin-3 expression by esophageal epithelial cells.

  • 430 different effects of proton pump inhibitors ppis on Eotaxin 3 protein expression by the proximal and distal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Introduction: PPIs can reduce esophageal eosinophilia, an effect usually attributed to decreased acid reflux. However, PPIs can suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal cells through mechanisms independent of effects on gastric acid. To explore the relative contributions of these acid-suppressive and acid-independent PPI actions, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid effects on Th2-cytokine-stimulated Eotaxin-3 expression by esophageal epithelial cells.Methods:We identified 10 children with initial esophageal biopsies showing ≥15 eos/hpf who had follow-up endoscopy after PPI treatment alone for ≥8 weeks. Eosinophils were quantified on HE Eotaxin-3 secretion was determined by ELISA. Results: For the group as a whole, mean maximum eos/hpf decreased with PPI treatment (50 to 40 eos/hpf); in 5 of the 10 patients, counts fell to <15 eos/hpf. There was strong correlation between maximum eos/hpf and maximum Eotaxin-3-positive epithelial cells/hpf (R=0.8332, P<0.0001). Although PPI effects in reducing acid reflux are strongest in the distal esophagus, Eotaxin-3 expression decreased significantly with PPIs only in the proximal esophagus (28 to 6 cells/hpf proximal, P=0.049; 14 to 24 cells/hpf distal esophagus, P=0.41). In vitro, EoE1-T and EoE2-T cells stimulated with IL-13 showed strong suppression of Eotaxin-3 secretion at pH4; weakly acidic pH levels had no consistent effects on Eotaxin3 in the same cell lines (Table). Treatment with omeprazole at weakly acidic pH did not suppress IL-13-stimulated Eotaxin-3 secretion as profoundly as exposure to pH4 alone (Table). Conclusions: In children with esophageal eosinophilia, PPIs significantly decrease expression of Eotaxin-3 protein in the proximal but not distal esophagus. In esophageal squamous cells in vitro, acid at pH4 decreases Eotaxin-3 secretion significantly. This in vitro finding might explain the disparate PPI effects on the proximal and distal esophagus. In the distal esophagus, where acid exposure that inhibits Eotaxin-3 secretion is greatest, suppression of acid reflux by PPIs might increase Eotaxin-3 secretion and mask acid-independent PPI suppressive effects on the protein. In the proximal esophagus, where acid exposure is least, acid-independent PPI effects in suppressing Eotaxin-3 secretion are manifest most greatly. Acid pH Effects on IL-13-Driven Eotaxin-3 Protein Secretion by Esophageal Squamous Cell Lines (EoE1-T and EoE2-T)

Xi Zhang - One of the best experts on this subject based on the ideXlab platform.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Eunice Odiase, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    Background & Aims In upper airway cells, T helper 2 cytokines that signal through interleukin-4 (IL-4) receptor-α have been shown to stimulate Eotaxin-3 secretion via a nongastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL-4 receptor-α signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by quantitative real-time polymerase chain reaction and Western blotting. IL-4–stimulated Eotaxin-3 secretion was measured by enzyme-linked immunosorbent assay after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester (calcium chelator), 2-aminoethoxydiphenyl borate (inhibitor of endoplasmic reticulum calcium release), verapamil, and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of patients with EoE and controls. Results EoE cells expressed ngH+,K+ATPase messenger RNA and protein. Omeprazole and SCH 28080 decreased IL-4–stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4–stimulated Eotaxin-3 secretion was blocked by ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester, 2-aminoethoxydiphenyl borate, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4–stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a nongastric proton pump that mediates T helper 2 cytokine–stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block T helper 2 cytokine–stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Eunice Odiase, Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    ABSTRACT Background & Aims In upper airway cells, Th2 cytokines that signal through IL-4 receptor alpha (IL4Rα) have been shown to stimulate Eotaxin-3 secretion via a non-gastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL4Rα signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by qPCR and western blotting. IL-4-stimulated Eotaxin-3 secretion was measured by ELISA after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), EGTA-AM (calcium chelator), 2-APB (inhibitor of endoplasmic reticulum calcium release), verapamil and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of EoE patients and controls. Results EoE cells expressed ngH+,K+ATPase mRNA and protein. Omeprazole and SCH 28080 decreased IL-4-stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4-stimulated Eotaxin-3 secretion was blocked by EGTA-AM, 2-APB, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4-stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a non-gastric proton pump that mediates Th2 cytokine-stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block Th2 cytokine-stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • jak stat6 pathway inhibitors block Eotaxin 3 secretion by epithelial cells and fibroblasts from esophageal eosinophilia patients promising agents to improve inflammation and prevent fibrosis in eoe
    2016
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Jason Y Park, Kathleen S Wilson, Rhonda F Souza
    Abstract:

    Background: Although most studies on treatments for eosinophilic esophagitis (EoE) have focused on effects in the epithelium, EoE is a transmural disease. Eosinophils that infiltrate the subepithelial layers of the esophagus lead to fibrosis and the serious complications of EoE, and current therapies have shown minimal effects on this fibrosis. We aimed to elucidate T helper (Th)2 cytokine effects on esophageal fibroblasts and to explore potential fibroblast-targeted therapies for EoE. Methods: We established telomerase-immortalized fibroblasts from human esophageal biopsies. We stimulated these esophageal fibroblasts with Th2 cytokines, and examined effects of omeprazole and inhibitors of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT6) pathway (AS1517499, leflunomide, and ruxolitinib) on STAT6 phosphorylation, STAT6 nuclear translocation, and Eotaxin-3 expression. We also measured the effects of these inhibitors in esophageal epithelial cells stimulated with Th2 cytokines. Results: As in esophageal epithelial cells, Th2 cytokines increased STAT6 phosphorylation, STAT6 nuclear translocation, Eotaxin-3 transcription and protein secretion in esophageal fibroblasts. Unlike in epithelial cells, however, omeprazole did not inhibit cytokine-stimulated Eotaxin-3 expression in fibroblasts. In contrast, JAK-STAT6 pathway inhibitors decreased cytokine-stimulated Eotaxin-3 expression in both fibroblasts and epithelial cells. Conclusions: Omeprazole does not inhibit Th2 cytokine-stimulated Eotaxin-3 expression by esophageal fibroblasts, suggesting that PPIs will have limited impact on subepithelial EoE processes such as fibrosis. JAK-STAT6 pathway inhibitors block Th2 cytokine-stimulated Eotaxin-3 expression both in fibroblasts and in epithelial cells, suggesting a potential role for JAK-STAT inhibitors in treating both epithelial inflammation and subepithelial fibrosis in EoE.

  • proton pump inhibitors decrease Eotaxin 3 expression in the proximal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Objective Besides reducing gastric acid secretion, proton pump inhibitors (PPIs) suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal epithelial cells through acid-independent, anti-inflammatory mechanisms. To explore acid-inhibitory and acid-independent, anti-inflammatory PPI effects in reducing esophageal eosinophilia, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid and bile salt effects on IL-13-stimulated Eotaxin-3 expression by esophageal epithelial cells.

  • 430 different effects of proton pump inhibitors ppis on Eotaxin 3 protein expression by the proximal and distal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Introduction: PPIs can reduce esophageal eosinophilia, an effect usually attributed to decreased acid reflux. However, PPIs can suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal cells through mechanisms independent of effects on gastric acid. To explore the relative contributions of these acid-suppressive and acid-independent PPI actions, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid effects on Th2-cytokine-stimulated Eotaxin-3 expression by esophageal epithelial cells.Methods:We identified 10 children with initial esophageal biopsies showing ≥15 eos/hpf who had follow-up endoscopy after PPI treatment alone for ≥8 weeks. Eosinophils were quantified on HE Eotaxin-3 secretion was determined by ELISA. Results: For the group as a whole, mean maximum eos/hpf decreased with PPI treatment (50 to 40 eos/hpf); in 5 of the 10 patients, counts fell to <15 eos/hpf. There was strong correlation between maximum eos/hpf and maximum Eotaxin-3-positive epithelial cells/hpf (R=0.8332, P<0.0001). Although PPI effects in reducing acid reflux are strongest in the distal esophagus, Eotaxin-3 expression decreased significantly with PPIs only in the proximal esophagus (28 to 6 cells/hpf proximal, P=0.049; 14 to 24 cells/hpf distal esophagus, P=0.41). In vitro, EoE1-T and EoE2-T cells stimulated with IL-13 showed strong suppression of Eotaxin-3 secretion at pH4; weakly acidic pH levels had no consistent effects on Eotaxin3 in the same cell lines (Table). Treatment with omeprazole at weakly acidic pH did not suppress IL-13-stimulated Eotaxin-3 secretion as profoundly as exposure to pH4 alone (Table). Conclusions: In children with esophageal eosinophilia, PPIs significantly decrease expression of Eotaxin-3 protein in the proximal but not distal esophagus. In esophageal squamous cells in vitro, acid at pH4 decreases Eotaxin-3 secretion significantly. This in vitro finding might explain the disparate PPI effects on the proximal and distal esophagus. In the distal esophagus, where acid exposure that inhibits Eotaxin-3 secretion is greatest, suppression of acid reflux by PPIs might increase Eotaxin-3 secretion and mask acid-independent PPI suppressive effects on the protein. In the proximal esophagus, where acid exposure is least, acid-independent PPI effects in suppressing Eotaxin-3 secretion are manifest most greatly. Acid pH Effects on IL-13-Driven Eotaxin-3 Protein Secretion by Esophageal Squamous Cell Lines (EoE1-T and EoE2-T)

Stuart J Spechler - One of the best experts on this subject based on the ideXlab platform.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Eunice Odiase, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    Background & Aims In upper airway cells, T helper 2 cytokines that signal through interleukin-4 (IL-4) receptor-α have been shown to stimulate Eotaxin-3 secretion via a nongastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL-4 receptor-α signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by quantitative real-time polymerase chain reaction and Western blotting. IL-4–stimulated Eotaxin-3 secretion was measured by enzyme-linked immunosorbent assay after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester (calcium chelator), 2-aminoethoxydiphenyl borate (inhibitor of endoplasmic reticulum calcium release), verapamil, and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of patients with EoE and controls. Results EoE cells expressed ngH+,K+ATPase messenger RNA and protein. Omeprazole and SCH 28080 decreased IL-4–stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4–stimulated Eotaxin-3 secretion was blocked by ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester, 2-aminoethoxydiphenyl borate, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4–stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a nongastric proton pump that mediates T helper 2 cytokine–stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block T helper 2 cytokine–stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Eunice Odiase, Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    ABSTRACT Background & Aims In upper airway cells, Th2 cytokines that signal through IL-4 receptor alpha (IL4Rα) have been shown to stimulate Eotaxin-3 secretion via a non-gastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL4Rα signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by qPCR and western blotting. IL-4-stimulated Eotaxin-3 secretion was measured by ELISA after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), EGTA-AM (calcium chelator), 2-APB (inhibitor of endoplasmic reticulum calcium release), verapamil and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of EoE patients and controls. Results EoE cells expressed ngH+,K+ATPase mRNA and protein. Omeprazole and SCH 28080 decreased IL-4-stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4-stimulated Eotaxin-3 secretion was blocked by EGTA-AM, 2-APB, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4-stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a non-gastric proton pump that mediates Th2 cytokine-stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block Th2 cytokine-stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • jak stat6 pathway inhibitors block Eotaxin 3 secretion by epithelial cells and fibroblasts from esophageal eosinophilia patients promising agents to improve inflammation and prevent fibrosis in eoe
    2016
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Jason Y Park, Kathleen S Wilson, Rhonda F Souza
    Abstract:

    Background: Although most studies on treatments for eosinophilic esophagitis (EoE) have focused on effects in the epithelium, EoE is a transmural disease. Eosinophils that infiltrate the subepithelial layers of the esophagus lead to fibrosis and the serious complications of EoE, and current therapies have shown minimal effects on this fibrosis. We aimed to elucidate T helper (Th)2 cytokine effects on esophageal fibroblasts and to explore potential fibroblast-targeted therapies for EoE. Methods: We established telomerase-immortalized fibroblasts from human esophageal biopsies. We stimulated these esophageal fibroblasts with Th2 cytokines, and examined effects of omeprazole and inhibitors of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT6) pathway (AS1517499, leflunomide, and ruxolitinib) on STAT6 phosphorylation, STAT6 nuclear translocation, and Eotaxin-3 expression. We also measured the effects of these inhibitors in esophageal epithelial cells stimulated with Th2 cytokines. Results: As in esophageal epithelial cells, Th2 cytokines increased STAT6 phosphorylation, STAT6 nuclear translocation, Eotaxin-3 transcription and protein secretion in esophageal fibroblasts. Unlike in epithelial cells, however, omeprazole did not inhibit cytokine-stimulated Eotaxin-3 expression in fibroblasts. In contrast, JAK-STAT6 pathway inhibitors decreased cytokine-stimulated Eotaxin-3 expression in both fibroblasts and epithelial cells. Conclusions: Omeprazole does not inhibit Th2 cytokine-stimulated Eotaxin-3 expression by esophageal fibroblasts, suggesting that PPIs will have limited impact on subepithelial EoE processes such as fibrosis. JAK-STAT6 pathway inhibitors block Th2 cytokine-stimulated Eotaxin-3 expression both in fibroblasts and in epithelial cells, suggesting a potential role for JAK-STAT inhibitors in treating both epithelial inflammation and subepithelial fibrosis in EoE.

  • proton pump inhibitors decrease Eotaxin 3 expression in the proximal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Objective Besides reducing gastric acid secretion, proton pump inhibitors (PPIs) suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal epithelial cells through acid-independent, anti-inflammatory mechanisms. To explore acid-inhibitory and acid-independent, anti-inflammatory PPI effects in reducing esophageal eosinophilia, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid and bile salt effects on IL-13-stimulated Eotaxin-3 expression by esophageal epithelial cells.

  • 430 different effects of proton pump inhibitors ppis on Eotaxin 3 protein expression by the proximal and distal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Introduction: PPIs can reduce esophageal eosinophilia, an effect usually attributed to decreased acid reflux. However, PPIs can suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal cells through mechanisms independent of effects on gastric acid. To explore the relative contributions of these acid-suppressive and acid-independent PPI actions, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid effects on Th2-cytokine-stimulated Eotaxin-3 expression by esophageal epithelial cells.Methods:We identified 10 children with initial esophageal biopsies showing ≥15 eos/hpf who had follow-up endoscopy after PPI treatment alone for ≥8 weeks. Eosinophils were quantified on HE Eotaxin-3 secretion was determined by ELISA. Results: For the group as a whole, mean maximum eos/hpf decreased with PPI treatment (50 to 40 eos/hpf); in 5 of the 10 patients, counts fell to <15 eos/hpf. There was strong correlation between maximum eos/hpf and maximum Eotaxin-3-positive epithelial cells/hpf (R=0.8332, P<0.0001). Although PPI effects in reducing acid reflux are strongest in the distal esophagus, Eotaxin-3 expression decreased significantly with PPIs only in the proximal esophagus (28 to 6 cells/hpf proximal, P=0.049; 14 to 24 cells/hpf distal esophagus, P=0.41). In vitro, EoE1-T and EoE2-T cells stimulated with IL-13 showed strong suppression of Eotaxin-3 secretion at pH4; weakly acidic pH levels had no consistent effects on Eotaxin3 in the same cell lines (Table). Treatment with omeprazole at weakly acidic pH did not suppress IL-13-stimulated Eotaxin-3 secretion as profoundly as exposure to pH4 alone (Table). Conclusions: In children with esophageal eosinophilia, PPIs significantly decrease expression of Eotaxin-3 protein in the proximal but not distal esophagus. In esophageal squamous cells in vitro, acid at pH4 decreases Eotaxin-3 secretion significantly. This in vitro finding might explain the disparate PPI effects on the proximal and distal esophagus. In the distal esophagus, where acid exposure that inhibits Eotaxin-3 secretion is greatest, suppression of acid reflux by PPIs might increase Eotaxin-3 secretion and mask acid-independent PPI suppressive effects on the protein. In the proximal esophagus, where acid exposure is least, acid-independent PPI effects in suppressing Eotaxin-3 secretion are manifest most greatly. Acid pH Effects on IL-13-Driven Eotaxin-3 Protein Secretion by Esophageal Squamous Cell Lines (EoE1-T and EoE2-T)

Qiuyang Zhang - One of the best experts on this subject based on the ideXlab platform.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Eunice Odiase, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    Background & Aims In upper airway cells, T helper 2 cytokines that signal through interleukin-4 (IL-4) receptor-α have been shown to stimulate Eotaxin-3 secretion via a nongastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL-4 receptor-α signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by quantitative real-time polymerase chain reaction and Western blotting. IL-4–stimulated Eotaxin-3 secretion was measured by enzyme-linked immunosorbent assay after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester (calcium chelator), 2-aminoethoxydiphenyl borate (inhibitor of endoplasmic reticulum calcium release), verapamil, and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of patients with EoE and controls. Results EoE cells expressed ngH+,K+ATPase messenger RNA and protein. Omeprazole and SCH 28080 decreased IL-4–stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4–stimulated Eotaxin-3 secretion was blocked by ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetoxymethyl ester, 2-aminoethoxydiphenyl borate, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4–stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a nongastric proton pump that mediates T helper 2 cytokine–stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block T helper 2 cytokine–stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • in esophageal squamous cells from eosinophilic esophagitis patients th2 cytokines increase Eotaxin 3 secretion through effects on intracellular calcium and a non gastric proton pump
    2021
    Co-Authors: Eunice Odiase, Qiuyang Zhang, Stuart J Spechler, Xi Zhang, Yan Chang, Melissa R Nelson, Uthra Balaji, Zui Pan, Rhonda F Souza
    Abstract:

    ABSTRACT Background & Aims In upper airway cells, Th2 cytokines that signal through IL-4 receptor alpha (IL4Rα) have been shown to stimulate Eotaxin-3 secretion via a non-gastric proton pump (ngH+,K+ATPase). To seek novel targets for eosinophilic esophagitis (EoE) treatments, we evaluated ngH+,K+ATPase expression in EoE squamous cells, and explored molecular pathways involved in Eotaxin-3 secretion by IL4Rα signaling. Methods ngH+,K+ATPase expression in EoE cells was evaluated by qPCR and western blotting. IL-4-stimulated Eotaxin-3 secretion was measured by ELISA after treatment with omeprazole, SCH 28080 (potassium-competitive acid blocker), EGTA-AM (calcium chelator), 2-APB (inhibitor of endoplasmic reticulum calcium release), verapamil and diltiazem (L-type calcium channel inhibitors). Intracellular calcium transients were measured by Fluo-4 fluorescence. Key experiments were confirmed in EoE primary cells and in RNA sequencing datasets from mucosal biopsies of EoE patients and controls. Results EoE cells expressed ngH+,K+ATPase mRNA and protein. Omeprazole and SCH 28080 decreased IL-4-stimulated Eotaxin-3 secretion. IL-4 increased intracellular calcium transients, and IL-4-stimulated Eotaxin-3 secretion was blocked by EGTA-AM, 2-APB, verapamil, and diltiazem. The combination of omeprazole and verapamil suppressed IL-4-stimulated Eotaxin-3 secretion more than either agent alone. EoE biopsies expressed higher ngH+,K+ATPase and exhibited more calcium signaling than controls. Conclusions EoE cells express a non-gastric proton pump that mediates Th2 cytokine-stimulated Eotaxin-3 secretion. IL-4 induces calcium release from the endoplasmic reticulum and calcium entry via L-type calcium channels, increasing intracellular calcium that contributes to Eotaxin-3 secretion by EoE cells. L-type calcium channel inhibitors block Th2 cytokine-stimulated Eotaxin-3 secretion, suggesting a potential role for these agents in EoE treatment.

  • jak stat6 pathway inhibitors block Eotaxin 3 secretion by epithelial cells and fibroblasts from esophageal eosinophilia patients promising agents to improve inflammation and prevent fibrosis in eoe
    2016
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Jason Y Park, Kathleen S Wilson, Rhonda F Souza
    Abstract:

    Background: Although most studies on treatments for eosinophilic esophagitis (EoE) have focused on effects in the epithelium, EoE is a transmural disease. Eosinophils that infiltrate the subepithelial layers of the esophagus lead to fibrosis and the serious complications of EoE, and current therapies have shown minimal effects on this fibrosis. We aimed to elucidate T helper (Th)2 cytokine effects on esophageal fibroblasts and to explore potential fibroblast-targeted therapies for EoE. Methods: We established telomerase-immortalized fibroblasts from human esophageal biopsies. We stimulated these esophageal fibroblasts with Th2 cytokines, and examined effects of omeprazole and inhibitors of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT6) pathway (AS1517499, leflunomide, and ruxolitinib) on STAT6 phosphorylation, STAT6 nuclear translocation, and Eotaxin-3 expression. We also measured the effects of these inhibitors in esophageal epithelial cells stimulated with Th2 cytokines. Results: As in esophageal epithelial cells, Th2 cytokines increased STAT6 phosphorylation, STAT6 nuclear translocation, Eotaxin-3 transcription and protein secretion in esophageal fibroblasts. Unlike in epithelial cells, however, omeprazole did not inhibit cytokine-stimulated Eotaxin-3 expression in fibroblasts. In contrast, JAK-STAT6 pathway inhibitors decreased cytokine-stimulated Eotaxin-3 expression in both fibroblasts and epithelial cells. Conclusions: Omeprazole does not inhibit Th2 cytokine-stimulated Eotaxin-3 expression by esophageal fibroblasts, suggesting that PPIs will have limited impact on subepithelial EoE processes such as fibrosis. JAK-STAT6 pathway inhibitors block Th2 cytokine-stimulated Eotaxin-3 expression both in fibroblasts and in epithelial cells, suggesting a potential role for JAK-STAT inhibitors in treating both epithelial inflammation and subepithelial fibrosis in EoE.

  • omeprazole blocks Eotaxin 3 expression by oesophageal squamous cells from patients with eosinophilic oesophagitis and gord
    2013
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Rhonda F Souza
    Abstract:

    Objective Eosinophilic oesophagitis (EoE) and gastro-oesophageal reflux disease (GORD) can have similar clinical and histological features. Proton pump inhibitors (PPIs) are used to distinguish the disorders, with the assumption that only GORD can respond to PPIs. Oesophageal expression of Eotaxin-3 stimulated by Th2 cytokines might contribute to oesophageal eosinophilia in EoE. Th2 cytokine effects on the oesophagus in GORD are not known. The objective of the authors was to explore the molecular mechanisms of Th2 cytokines on Eotaxin-3 expression by oesophageal squamous cells from patients with GORD and EoE, and the effects of omeprazole on that Eotaxin-3 expression. Design Using telomerase-immortalised and primary cultures of oesophageal squamous cells from GORD and EoE patients, the authors measured Eotaxin-3 protein secretion stimulated by Th2 cytokines (interleukin (IL)-4 and IL-13). Eotaxin-3 promoter constructs were used to study transcriptional regulation. Cytokine-induced Eotaxin-3 mRNA and protein expression were measured in the presence or absence of omeprazole. Results There were no significant differences between EoE and GORD primary cells in cytokine-stimulated Eotaxin-3 protein secretion levels. In EoE and GORD cell lines, IL-4 and IL-13 activated the Eotaxin-3 promoter, and significantly increased Eotaxin-3 mRNA and protein expression. Omeprazole blocked the cytokine-stimulated increase in Eotaxin-3 mRNA and protein expression in EoE and GORD cell lines. Conclusion Oesophageal squamous cells from GORD and EoE patients express similar levels of Eotaxin-3 when stimulated by Th2 cytokines, and omeprazole blocks that Eotaxin-3 expression. These findings suggest that PPIs might have eosinophil-reducing effects independent of effects on acid reflux and that response to PPIs might not distinguish EoE from GORD.

  • omeprazole blocks stat6 binding to the Eotaxin 3 promoter in eosinophilic esophagitis cells
    2012
    Co-Authors: Xi Zhang, Edaire Cheng, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xiaofang Huo, Thai H Pham, Rhonda F Souza
    Abstract:

    Background Patients who have esophageal eosinophilia without gastroesophageal reflux disease (GERD) nevertheless can respond to proton pump inhibitors (PPIs), which can have anti-inflammatory actions independent of effects on gastric acid secretion. In esophageal cell cultures, omeprazole has been reported to inhibit Th2 cytokine-stimulated expression of Eotaxin-3, an eosinophil chemoattractant contributing to esophageal eosinophilia in eosinophilic esophagitis (EoE). The objective of this study was to elucidate molecular mechanisms underlying PPI inhibition of IL-4-stimulated Eotaxin-3 production by esophageal cells.

Edaire Cheng - One of the best experts on this subject based on the ideXlab platform.

  • jak stat6 pathway inhibitors block Eotaxin 3 secretion by epithelial cells and fibroblasts from esophageal eosinophilia patients promising agents to improve inflammation and prevent fibrosis in eoe
    2016
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Jason Y Park, Kathleen S Wilson, Rhonda F Souza
    Abstract:

    Background: Although most studies on treatments for eosinophilic esophagitis (EoE) have focused on effects in the epithelium, EoE is a transmural disease. Eosinophils that infiltrate the subepithelial layers of the esophagus lead to fibrosis and the serious complications of EoE, and current therapies have shown minimal effects on this fibrosis. We aimed to elucidate T helper (Th)2 cytokine effects on esophageal fibroblasts and to explore potential fibroblast-targeted therapies for EoE. Methods: We established telomerase-immortalized fibroblasts from human esophageal biopsies. We stimulated these esophageal fibroblasts with Th2 cytokines, and examined effects of omeprazole and inhibitors of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT6) pathway (AS1517499, leflunomide, and ruxolitinib) on STAT6 phosphorylation, STAT6 nuclear translocation, and Eotaxin-3 expression. We also measured the effects of these inhibitors in esophageal epithelial cells stimulated with Th2 cytokines. Results: As in esophageal epithelial cells, Th2 cytokines increased STAT6 phosphorylation, STAT6 nuclear translocation, Eotaxin-3 transcription and protein secretion in esophageal fibroblasts. Unlike in epithelial cells, however, omeprazole did not inhibit cytokine-stimulated Eotaxin-3 expression in fibroblasts. In contrast, JAK-STAT6 pathway inhibitors decreased cytokine-stimulated Eotaxin-3 expression in both fibroblasts and epithelial cells. Conclusions: Omeprazole does not inhibit Th2 cytokine-stimulated Eotaxin-3 expression by esophageal fibroblasts, suggesting that PPIs will have limited impact on subepithelial EoE processes such as fibrosis. JAK-STAT6 pathway inhibitors block Th2 cytokine-stimulated Eotaxin-3 expression both in fibroblasts and in epithelial cells, suggesting a potential role for JAK-STAT inhibitors in treating both epithelial inflammation and subepithelial fibrosis in EoE.

  • proton pump inhibitors decrease Eotaxin 3 expression in the proximal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Objective Besides reducing gastric acid secretion, proton pump inhibitors (PPIs) suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal epithelial cells through acid-independent, anti-inflammatory mechanisms. To explore acid-inhibitory and acid-independent, anti-inflammatory PPI effects in reducing esophageal eosinophilia, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid and bile salt effects on IL-13-stimulated Eotaxin-3 expression by esophageal epithelial cells.

  • 430 different effects of proton pump inhibitors ppis on Eotaxin 3 protein expression by the proximal and distal esophagus of children with esophageal eosinophilia
    2014
    Co-Authors: Jason Y Park, Rhonda F Souza, Stuart J Spechler, Xi Zhang, Nathalie Nguyen, Edaire Cheng
    Abstract:

    Introduction: PPIs can reduce esophageal eosinophilia, an effect usually attributed to decreased acid reflux. However, PPIs can suppress Th2-cytokine-stimulated expression of an eosinophil chemoattractant (Eotaxin-3) by esophageal cells through mechanisms independent of effects on gastric acid. To explore the relative contributions of these acid-suppressive and acid-independent PPI actions, we studied Eotaxin-3 expression by the proximal and distal esophagus of children with esophageal eosinophilia before and after PPI therapy. In vitro, we studied acid effects on Th2-cytokine-stimulated Eotaxin-3 expression by esophageal epithelial cells.Methods:We identified 10 children with initial esophageal biopsies showing ≥15 eos/hpf who had follow-up endoscopy after PPI treatment alone for ≥8 weeks. Eosinophils were quantified on HE Eotaxin-3 secretion was determined by ELISA. Results: For the group as a whole, mean maximum eos/hpf decreased with PPI treatment (50 to 40 eos/hpf); in 5 of the 10 patients, counts fell to <15 eos/hpf. There was strong correlation between maximum eos/hpf and maximum Eotaxin-3-positive epithelial cells/hpf (R=0.8332, P<0.0001). Although PPI effects in reducing acid reflux are strongest in the distal esophagus, Eotaxin-3 expression decreased significantly with PPIs only in the proximal esophagus (28 to 6 cells/hpf proximal, P=0.049; 14 to 24 cells/hpf distal esophagus, P=0.41). In vitro, EoE1-T and EoE2-T cells stimulated with IL-13 showed strong suppression of Eotaxin-3 secretion at pH4; weakly acidic pH levels had no consistent effects on Eotaxin3 in the same cell lines (Table). Treatment with omeprazole at weakly acidic pH did not suppress IL-13-stimulated Eotaxin-3 secretion as profoundly as exposure to pH4 alone (Table). Conclusions: In children with esophageal eosinophilia, PPIs significantly decrease expression of Eotaxin-3 protein in the proximal but not distal esophagus. In esophageal squamous cells in vitro, acid at pH4 decreases Eotaxin-3 secretion significantly. This in vitro finding might explain the disparate PPI effects on the proximal and distal esophagus. In the distal esophagus, where acid exposure that inhibits Eotaxin-3 secretion is greatest, suppression of acid reflux by PPIs might increase Eotaxin-3 secretion and mask acid-independent PPI suppressive effects on the protein. In the proximal esophagus, where acid exposure is least, acid-independent PPI effects in suppressing Eotaxin-3 secretion are manifest most greatly. Acid pH Effects on IL-13-Driven Eotaxin-3 Protein Secretion by Esophageal Squamous Cell Lines (EoE1-T and EoE2-T)

  • omeprazole blocks Eotaxin 3 expression by oesophageal squamous cells from patients with eosinophilic oesophagitis and gord
    2013
    Co-Authors: Edaire Cheng, Chunhua Yu, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xi Zhang, Rhonda F Souza
    Abstract:

    Objective Eosinophilic oesophagitis (EoE) and gastro-oesophageal reflux disease (GORD) can have similar clinical and histological features. Proton pump inhibitors (PPIs) are used to distinguish the disorders, with the assumption that only GORD can respond to PPIs. Oesophageal expression of Eotaxin-3 stimulated by Th2 cytokines might contribute to oesophageal eosinophilia in EoE. Th2 cytokine effects on the oesophagus in GORD are not known. The objective of the authors was to explore the molecular mechanisms of Th2 cytokines on Eotaxin-3 expression by oesophageal squamous cells from patients with GORD and EoE, and the effects of omeprazole on that Eotaxin-3 expression. Design Using telomerase-immortalised and primary cultures of oesophageal squamous cells from GORD and EoE patients, the authors measured Eotaxin-3 protein secretion stimulated by Th2 cytokines (interleukin (IL)-4 and IL-13). Eotaxin-3 promoter constructs were used to study transcriptional regulation. Cytokine-induced Eotaxin-3 mRNA and protein expression were measured in the presence or absence of omeprazole. Results There were no significant differences between EoE and GORD primary cells in cytokine-stimulated Eotaxin-3 protein secretion levels. In EoE and GORD cell lines, IL-4 and IL-13 activated the Eotaxin-3 promoter, and significantly increased Eotaxin-3 mRNA and protein expression. Omeprazole blocked the cytokine-stimulated increase in Eotaxin-3 mRNA and protein expression in EoE and GORD cell lines. Conclusion Oesophageal squamous cells from GORD and EoE patients express similar levels of Eotaxin-3 when stimulated by Th2 cytokines, and omeprazole blocks that Eotaxin-3 expression. These findings suggest that PPIs might have eosinophil-reducing effects independent of effects on acid reflux and that response to PPIs might not distinguish EoE from GORD.

  • omeprazole blocks stat6 binding to the Eotaxin 3 promoter in eosinophilic esophagitis cells
    2012
    Co-Authors: Xi Zhang, Edaire Cheng, Qiuyang Zhang, Stuart J Spechler, David H Wang, Xiaofang Huo, Thai H Pham, Rhonda F Souza
    Abstract:

    Background Patients who have esophageal eosinophilia without gastroesophageal reflux disease (GERD) nevertheless can respond to proton pump inhibitors (PPIs), which can have anti-inflammatory actions independent of effects on gastric acid secretion. In esophageal cell cultures, omeprazole has been reported to inhibit Th2 cytokine-stimulated expression of Eotaxin-3, an eosinophil chemoattractant contributing to esophageal eosinophilia in eosinophilic esophagitis (EoE). The objective of this study was to elucidate molecular mechanisms underlying PPI inhibition of IL-4-stimulated Eotaxin-3 production by esophageal cells.