Concentrations of IL-6 and IL-1β (BD Biosciences) in culture supernatants were measured with ELISAs according to the manufacturer’s recommendations.
C. rodentium–specific Ig amounts in sera and fecal were measured with SBA Clonotyping System (Southern Biotech) according to the manufacturer’s instructions. C. rodentium was grown overnight in LB broth with shaking at 37°C. After centrifugation, bacterial pellets were resuspended in sterile PBS and sonicated on ice. The supernatant was collected by removing the pellet after centrifugation at 13,000 rpm for 5 min and used to coat ELISA plates at the concentration of 5 µg/ml. Feces were collected from individual mice and then weighed and homogenized in sterile PBS at the concentration of 50 mg/ml. After centrifugation at 13,000 rpm for 5 min, the supernatants were collected and frozen at −80°C.
C. rodentium–specific Ig amounts in sera and fecal were measured with SBA Clonotyping System (Southern Biotech) according to the manufacturer’s instructions. C. rodentium was grown overnight in LB broth with shaking at 37°C. After centrifugation, bacterial pellets were resuspended in sterile PBS and sonicated on ice. The supernatant was collected by removing the pellet after centrifugation at 13,000 rpm for 5 min and used to coat ELISA plates at the concentration of 5 µg/ml. Feces were collected from individual mice and then weighed and homogenized in sterile PBS at the concentration of 50 mg/ml. After centrifugation at 13,000 rpm for 5 min, the supernatants were collected and frozen at −80°C.