Cytokine analyses
Cytokine concentrations were determined in untreated lavage fluids.IL-17A, IL-17F and TGF-β1 concentrations were measured with commercial ELISA kits (Gen-Probe Diaclone, France) and IL-17E concentrations was determined by using Human IL-17E ELISA Construction Kit (Antigenix America). IL-6 and IL-10 concentrations were measured using high-sensitivity ELISA kits, R&D Systems (Abingdon United Kingdom), which the lower limit detection for IL-6 was 0.007–0.090 (mean 0.031) and for IL-10 was 0.03–0.17 (mean 0.09) for IL-10. Here a lower limit of detection for both IL-6 and IL-10 have been determined by adding 3SD of O.D. blank value. The calculated concentrations below the lower standard point, but above the sensitity to detect minimum dose, recommended in the kit, were accepted as valid values.