| OCR Text |
Show 49 excluded from further analysis, which included one patient with autoimmune esophagitis, one patient with Crohn’s disease, one patient with systemic scleroderma, and one patient with severe allergic asthma. Esophageal secretions were collected by endoscopic cytology brush prior to biopsy. Patients were diagnosed with EoE by standard criteria [1], 15 eosinophils per high powered field in two or more biopsies. 5.3.2 Collection of esophageal secretions Esophageal secretions were collected prior to esophageal biopsies during upper endoscopy. A Cook Medical cytobrush was passed through the endoscope and applied to the lumen of the esophagus withdrawn from the distal 5cm to the mid esophagus (≈10cm). The brushing was repeated until the bristles demonstrated saturation via endoscopic view of secretions from the esophagus. The brush was then removed and the frozen at -70 degrees Celsius until analysis. Detection of food triggers was performed clinically in active EoE patients who opted for food elimination. Food triggers were identified via elimination diet with 1) a single food elimination resulting in improvement in symptoms and esophageal biopsies with less than 15 eosinophils/HPF or 2) via multiple food elimination diets (i.e., two, four or six food or more) with resolution of esophageal eosinophilia to less than 15 eosinophils/HPF and recurrence of eosinophilia on biopsies greater than 15 eosinophils/HPF with 4 weeks of specific food reintroduction. All implicated foods resulting in recurrence of EoE were documented in the patient chart. 5.3.3 ImmunoCAP profiling of food-specific antibodies Brushes were thawed and centrifuged for 1 minute at 10,000RPM, yielding 21-23 microliters of fluid per brush. Fluids derived from esophageal brushings (≈20µL of esophageal secretions) were diluted in 1 mL Phadia diluent (1:50) (Product No. 10-9498-01, Kalamazoo, MI), and incubated for 30 minutes prior to IgG4 and IgA testing. Phadia 100 ImmunoCAP system (ThermoFisher Scientific, Kalamazoo, MI) was used to measure the IgG4 and IgA antibodies to four food allergens: wheat f1, soybean f14, casein f78, and egg f245. Briefly, approximately 40µL of diluted solution were added to the food specific CAPs, followed by an incubation of IgA/IgG4 conjugate and addition of development solution. The reaction was stopped by adding the stop solution, and the absorbance (response value) are measured by the spectrophotometer within the Phadia 100 system. Calibration curves |
