10 research outputs found
Infiltrating eosinophils express HR and correlate with mucosal HR expression in active EoE.
<p>Immunofluorescent localization of MBP and H1R (<b>A-C</b>), H2R (<b>D-F</b>) and H4R (<b>G-I</b>) in esophageal biopsies from active EoE subject (original magnification 200x). Arrows indicate MBP-HR colocalization. Representative cells with MBP-HR colocalization are magnified in inserts. Correlation of H1R (<b>J</b>), H2R (<b>K</b>), and H4R (<b>L</b>) mRNA in human esophageal biopsies to peak tissue eosinophil count per hpf, using Spearman correlation and linear regression analyses.</p
H1R and H2R are expressed in human esophageal mucosal biopsies.
<p>Quantification of HR expression in esophageal biopsies from nonEoE control biopsies (<b>A</b>). Kruskal wallis test and Dunn’s multiple test was used to determine significance between groups. Horizontal lines represent medians of each data set (***p<0.001). Immunofluorescence for H1R (<b>B</b>), H2R (<b>C</b>), and H4R (<b>D</b>) in esophagus of non-EoE control, subjects (original magnification 200x).</p
Expression of H1R, H2R, and H4R 4 are increased in esophageal biopsies in actively inflamed EoE.
<p>Comparison of H1,H2, H3, and H4 receptor expression in non-EoE control, inactive EoE and active EoE biopsies, using Mann-Whitney test (<b>A-D</b>). Horizontal bars represent median of each data set. Horizontal lines represent median of the data set.</p
Esophageal epithelial cells produce cytokines in response to histamine stimulation <i>in vitro</i>.
<p>Quantification of GM-CSF, TNFα, and IL-8 (pg/mL) secretion by histamine stimulated EPC2-hTERT cells (100uM histamine for 24 hrs). Two-tailed student t-test was used for statistical analysis. **p<0.005,***<0.0005. Effect of pretreatment with H1R antagonist pyrilamine or H2R antagonists (cimetidine and ranitidine) prior to histamine stimulation upon cytokine secretion in EPC2-hTERT cells (<b><i>B-D</i></b>). One way ANOVA with Bonferroni post-test was used for statistical analysis. *p<0.05, **p<.01,****p<0.0001. ns = not significant. Data are represented as the mean +/- SEM of n = 6.</p
Primary and immortalized esophageal epithelial cells express H1R and H2R.
<p>HR mRNA expression in human esophageal epithelial cell line, EPC2-hTERT (<b>A</b>) and primary esophageal epithelial cell lines derived from non-EoE control (<b>B,D,E</b>) and EoE (<b>C,D,E</b>) subjects. <b>A</b>: Data represent mean +/- SEM for n = 3. <b>B-E</b>: horizontal bars represent median with IQR. Mann-Whitney test was employed to quantify significance. *p<0.05, ns = not significant.</p
Preferential Secretion of Thymic Stromal Lymphopoietin (TSLP) by Terminally Differentiated Esophageal Epithelial Cells: Relevance to Eosinophilic Esophagitis (EoE) - Fig 2
<p><b>Calcium induces terminal differentiation of esophageal epithelial cells in-vitro A)</b> Cytokeratin 13 (CK13) and, <b>B)</b> involucrin (IVL), mRNA expression in response to high calcium (1.8mM CaCl<sub>2</sub>) media at 0, 24, 48, and 72 hours. Results are representative of three separate experiments. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001 as compared to undifferentiated (0 hr) cells. Immunoblot for <b>C)</b> involucrin in calcium differentiated EPC2-hTERT cells. <b>D</b>) IHC for TSLP on organotypic cultures of esophageal epithelium. SSE = Stratified squamous epithelium; B = Basal epithelium, Sub = subepithelium.</p
Patient demographics, and allergy test results.
<p>Patient demographics, and allergy test results.</p
Food antigens induce esophageal epithelial TSLP expression.
<p>Quantification of TSLP mRNA expression following food antigen stimulation in undifferentiated <b>(A)</b> and differentiated <b>(B)</b> EPC2-hTERT cells. <b>C</b>) TSLP secretion (pg/mL) was quantified in undifferentiated and differentiated EPC2-hTERT cells. TSLP secretion in differentiated primary esophageal epithelial cells from non-EoE control subjects (N = 7) <b>(D)</b> and EoE subjects (N = 20) <b>(E)</b> following stimulation with food antigens. All cell lines were stimulated for 24 hours. (<b>F</b>) comparison of ova mediated TSLP expression in differentiated inactive and active EoE cells. The dashed lines represent the lower limit of sensitivity of the ELISA kit (8pg/mL). Bars represent the mean of each data set. * p<0.05, **p<0.01, ***p <0.001 as compared to unstimulated controls.</p
TSLP expression is restricted to the suprabasal differentiated compartment of the esophageal epithelium.
<p>Immunohistochemistry for TSLP (A,C,E) and cytokeratin 13 (CK13) (B,D,F) in esophageal pinch biopsies from representative non-EoE control subjects, subjects with inactive EoE (<15 eosinophils per hpf), and active EoE (≥15 eosinophils per hpf). 200X magnification. Larger images represent 100X magnification. The box represents the area chosen for 200X magnification in the smaller side panel. SSE = Stratified squamous epithelium; B = Basal epithelium.</p
Terminal differentiation of human esophageal epithelial cells enhances the inducible expression and secretion of TSLP protein <i>in vitro</i>.
<p>TSLP mRNA expression in poly (I:C)-stimulated undifferentiated (<b>A</b>) and differentiated (<b>B</b>) EPC2-hTERT cells. <b>C)</b> TSLP protein secretion (pg/mL) by poly (I:C)-stimulated undifferentiated and differentiated EPC2-hTERT cells. <b>D</b>) TLR3 mRNA expression in undifferentiated and differentiated EPC2-hTERT cells. Results are representative of three separate experiments. * p<0.05, p<0.01***p <0.001, p<0.0001 as compared to undifferentiated cells.</p