Clone rb6 8c5

IL-4, IL-5, IL-13, IgE and IgG1 in BALF were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits from eBioscience according to the manufacturer’s recommendations. Flow cytometry was used to test the percentages of the various inflammatory cells in the BAL. Eosinophils were defined as SSC^high SiglecF⁺ (clone E50-2440, BD) Mac-3⁻ (M3/84, BD) cells, alveolar macrophages were identified as SSC^high SiglecF⁺ Mac-3⁺ cells, granulocytes were recognized as SSC^high Gr-1⁺ (clone RB6-8C5, eBioscience) cells, and lymphocytes were identified as FSC^low/SSC^low and expressing CD3 (clone 145-2C11, eBioscience) or CD19 (clone 1D3, eBioscience).

Pancreatic cells isolated from each group were washed by a fresh FACS buffer for three times, then cell suspensions were treated with the anti-mouse CD16/CD32 (1:100 ratio, clone 93, eBioscience, Santa Clara, CA, USA) for 15 min to block the Fc receptor. Cell suspensions were subsequently labeled with anti-mouse PE-Cyanine 5 conjugated CD11b (1:500 ratio, clone M1/70, eBioscience, Santa Clara, CA, USA) and anti-mouse FITC conjugated F4/80 (1:500 ratio, clone BM8, eBioscience, Santa Clara, CA, USA), and/or anti-mouse PE conjugated Ly6G (1:500 ratio, clone RB6-8C5, eBioscience, Santa Clara, CA, USA) for 30 min on ice in the dark to detect macrophages or neutrophils. Stained cells were measured with the FACS Calibur (Becton Dickinson, San Jose, CA, USA) and flow cytometric data were analyzed using CellQuestPro software.