Monoclonal antibody cd11b

Gut antigen presenting cells were isolated using techniques described previously elsewhere.⁴ (link) Briefly, small intestines were removed, opened longitudinally to flush out feces, cut into 15-mm pieces, and then incubated for 20 minutes at 37°C on a shaker in Hank’s balanced salt solution supplemented with 5% heat-inactivated fetal bovine serum and 2 mM EDTA. After passing the preparation through a metal filter, the intestinal fragments were collected, and the step was repeated. Subsequently, intestinal fragments were minced and incubated for 20 minutes at 37°C on a shaker in Hank’s balanced salt solution supplemented with 5% heat-inactivated fetal bovine serum and 1 mg/mL type VIII collagenase (Sigma-Aldrich, St. Louis, MO). The cell suspension was passed through a cell strainer to remove debris, washed, and then the CD11b⁺ cells were enriched with a CD11b MACS kit (Miltenyi Biotec, Bergisch Gladbach, Germany). In some instances, intestinal macrophages were labeled using monoclonal antibody CD11b (BD Biosciences, San Jose, CA) and sorted using a FACS Vantage (BD Biosciences). The percentage of macrophages in cell population was examined by staining with antibody specific for the macrophage marker F4/80.