Age- and sex-matched non-obese diabetic (NOD) mice born and bred within our animal facility were used for all experiments. Mice were housed in OptiMouse cages with corn cob bedding and up to 5 cage-mates. Mice had ad libitum access to chow and reverse osmosis chlorinated (2-3ppm)-purified water. Housing rooms were kept on a 14.5/9.5-hour light/dark cycle with temperature maintained at 22-25°C and 50-70% humidity. Sentinel mice were placed on dirty bedding and nesting material in experimental rooms and subsequently tested on a quarterly basis for presence of parasites (pinworms and fur/follicular mites, Pneumocystis spp. (carinii, murina)), bacteria (Mycoplasma pulmonis), and viruses (RADIL Comprehensive Panel) including Mouse Hepatitis Virus (MHV), Mouse Minute Virus (MMV), Mouse Parvovirus (NS1 – Generic Parvovirus & MPV 1-5), Theiler’s Murine Encephalitis Virus (TMEV), Epizootic Diarrhea of Infant Mice (EDIM), Sendai Virus, Pneumonia Virus of Mice (PVM), Reo3 virus (REO3), Lymphocytic Choriomeningitis Virus (LCMV), Ectromelia virus, Murine Adenovirus I/II (MAV1/MAV11), and Polyomavirus. All experiments were performed in compliance with protocols approved by the Animal Care Committee of The University of British Columbia.
Virus stocks for coxsackievirus B4 Edwards strain 2 (CVB4) were prepared as previously described (15 (link)). Normoglycemic NOD mice at 11-12 weeks old were injected intraperitoneally with either 400 plaque-forming units (pfu) CVB4 or DMEM vehicle. Non-fasting blood glucose was monitored using a OneTouch LifeScan monitor. Mice were considered diabetic with two consecutive readings above 16.2 mg/dL separated by 24 hours.
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