Animals and Bacteria

Specific pathogen-free (SPF) BALB/c mice, female, 5–6 weeks of age, 14 ± 2 g, were purchased from the Experimental Animal Center of Hubei University of Medicine. This study was approved by the Animal Ethical and Experimental Committee of Hubei University of Medicine.

The mouse-adapted H. pylori strain SS1 was obtained from the National Center for Disease Control and Prevention. H. pylori was cultured on Columbia blood agar plates enriched with 10% defibrinated horse blood, polymyxin B (161.5 µg/mL), vancomycin (10 µg/mL), trimethoprim (5 µg/mL), and amphotericin B (2.5 µg/mL) under microaerobic conditions (5% O₂, 10% CO₂, and 85% N₂) at 37°C for 3–5 days.

Helicobacter pylori lysate preparation: H. pylori SS1 were harvested from the plates and suspended in 0.01 M PBS. The suspension mixture was then pulse sonicated (Sonics, USA) for 5 min at 20% capacity while kept in an ice bath. H. pylori lysate was snap frozen in liquid nitrogen and kept at −80°C until use.