H. pylori bacterial culture

J166 and 7.13 are two H. pylori strains, CagA + types. Both were used for the in vitro studies. J166 H. pylori is a clinical isolate of human-derived H. pylori [29], and 7.13 H. pylori is derived through in vivo adaptation of a clinical H. pylori strain, B128 [30]. For the mouse infection, we utilized the pre-mouse Sydney strain 1 (PMSS1). PMSS1 is the wild-type rodent-adapted cag + H. pylori strain derived from the parental strain of the mouse Sydney strain (SS1), acquired form a clinical isolate of a duodenal ulcer patient [31]. All the H. pylori strains were a gift from Dr. Peek at Vanderbilt University. The cultures of H. pylori were made on Brucella agar (BBL/Becton Dickinson, Sparks, MD) supplemented with 5% heat-inactivated BSA (Invitrogen) and a combination of antibiotics (vancomycin cat#1404-93-9, 100 µg/ml; bacitracin cat# 1405-87-4, 200 µg/ml; amphotericin B cat#1397-89-3, 20 µg/ml; nalidixic acid cat#389-08-2, 10.7 µg/ml; polymyxin B cat#1405-20-5, 3.3 µg/ml) (Sigma-Aldrich, St. Louis, MO). H. pylori cultures from inoculation were grown in Brucella broth supplemented with 5% BSA and vancomycin antibiotic. After 24 h, bacteria were pelleted and resuspended in Brucella broth.