Increased HLA-DR expression on tissue eosinophils in eosinophilic esophagitis.
ABSTRACT The aim of the study was to investigate whether eosinophils have increased human leukocyte antigen (HLA)-DR expression in subjects with eosinophilic esophagitis (EoE) compared with controls.
Patients who were undergoing an upper endoscopy with biopsies for suspected gastroesophageal reflux disease (GERD) or EoE at Lucile Packard Children's Hospital were enrolled. In total, the blood and tissue samples of 10 healthy controls (HC), 11 subjects with GERD, and 10 with EoE were studied. Multiple tissue staining to identify eosinophils (via eosinophil cationic protein-clone EG2) and major histocompatibility complex class II cell surface receptors (via HLA-DR) was performed via immunohistochemistry. The peripheral blood was analyzed using flow cytometry to detect eosinophil HLA-DR expression among these subjects.
In the tissue, a greater proportion of eosinophils expressed HLA-DR among the subjects with EoE (mean 0.83 +/- 0.14, n = 9) relative to those with GERD (mean 0.18 +/- 0.19, n = 8, P < 0.01) and HC (mean 0.18 +/- 0.13, n = 6, P < 0.01). In total, 6 participants (4 HC subjects and 2 subjects with GERD) did not have any eosinophils identified on tissue staining and were unable to be included in the present statistical analysis. In the blood, there was no statistically significant difference in eosinophil HLA-DR expression among HC subjects (mean 415 +/- 217, n = 6), subjects with GERD (mean 507 +/- 429, n = 2), and those with EoE (mean 334 +/- 181, n = 6).
These data demonstrate that the eosinophils from the esophagus of subjects with EoE have increased HLA-DR expression within this tissue.
- SourceAvailable from: PubMed Central[Show abstract] [Hide abstract]
ABSTRACT: The aim of this study is to explore the localization of human mesenchymal stem cells from umbilical cord matrix (hMSCs-UC) and the role of these cells in the repair of foot ulcerate tissue in diabetic foot ulcers in rats. A diabetic rat model was established by administering Streptozotocin. Diabetic foot ulceration was defined as non-healing or delayed-healing of empyrosis on the dorsal hind foot after 14 weeks. hMSCs-UC were delivered through the left femoral artery. We evaluated the localization of hMSCs-UC and their role in tissue repair in diabetic foot ulcers by histological analysis, PCR, and immunohistochemical staining. A model for diabetes was established in 54 out of 60 rats (90% success rate) and 27 of these rats were treated with hMSCs-UC. The area of ulceration was significantly and progressively reduced at 7 and 14 days following treatment with hMSCs-UC. This gross observation was strongly supported by the histological changes, including newly developed blood vessels and proliferation of inflammatory cells at 3 days post-treatment, significant increase in granulation tissue at 7 days post-treatment and squamous epithelium or stratified squamous epithelium at 14 days post-treatment. Importantly, human leukocyte antigen type-I (HLA-1) was confirmed in ulcerated tissue by RT-PCR. The expression of cytokeratin 19 was significantly increased in diabetic model rats, with no detectable change in cytokeratin 10. Additionally, both collagens I and III increased in model rats treated with hMSCs-UC, but the ratio of collagen I/III was less significant in treated rats compared with control rats. These results suggest that hMSCs-UC specifically localize to the target ulcerated tissue and may promote the epithelialization of ulcerated tissue by stimulating the release of cytokeratin 19 from keratinocytes and extracellular matrix formation.International journal of biological sciences 01/2013; 10(1):80-9. · 4.37 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: OBJECTIVES:: Evidence suggests eosinophils may be acting as antigen presenting cells (APC) by presenting antigen to T-cells. We investigated the surface proteins of eosinophils and T-cells in the esophageal biopsies of eosinophilic eosphagitis (EoE) patients, gastroeosphageal reflux (GERD) patients, and healthy controls. METHODS:: Subjects were categorized as EoE, GERD or healthy control (HC). In esophageal tissue, EG2+ eosinophils were stained for the antigen presenting cell markers, CD40 or CD80, via immunohistochemistry. CD3+ T-cells were stained for co-stimulatory markers CD40L or CD28, and for activation markers, CD69 or CD134 via immunofluorescence or immunohistochemistry. RESULTS:: Eosinophils stained with CD40 and CD80. The number of EG2+CD40+ cells was increased in EoE (mean = 19.1 ± 14.8 cells/HPF, n = 11), in comparison to GERD (mean = 0.13 ± 0.19 cells/HPF, n = 5, p < 0.01) and HC (mean = 0.3 ± 0.7 cells/HPF, n = 5, p < 0.01). There was an elevation in EG2+CD80+ cells in EoE (mean = 18.1 ± 16.2 cells/HPF, n = 10), GERD (mean = 1.7 ± 2.8 cells/HPF, n = 6, p < 0.01) or HC (mean = 0.8 ± 1.3 cells/HPF, n = 6, p < 0.01). CD3+ T-cells stained with CD40L (not quantified). CD3+ T-cells stained with CD28 at elevated levels in EoE (mean = 14 ± 8.7 cells/HPF, n = 9), versus GERD (mean = 3.3 ± 1.2 cells/HPF, n = 6, p < 0.05) or HC (mean = 3.0 ± 3.2 cells/HPF, n = 7, p < 0.01). The number of CD3+CD69+ cells was highest in EoE (mean = 14.8 ± 7.5 cells/HPF, n = 6) versus GERD (mean = 0.8 ± 0.9 cells/HPF, n = 6, p < 0.001) or HC (mean = 2.7 ± 2.5 cells/HPF, n = 6, p < 0.001). CONCLUSIONS:: We show that esophageal eosinophils express CD40 and CD80, and T-cells with CD40L, CD28 and CD69. The number of double stained cells was higher in EoE in comparison to controls groups. These data support the hypothesis that eosinophils in EoE might act as APC, activating T-cells.Journal of pediatric gastroenterology and nutrition 10/2012; · 2.18 Impact Factor
- Journal of pediatric gastroenterology and nutrition 11/2012; · 2.18 Impact Factor