After nine administrations of bacteria or vehicle, mice (11 weeks old) were immunised with bovine type II collagen (Chondrex, Inc., Redmond, WA) emulsified in complete Freund’s adjuvant (Chondrex) at a final concentration of 0.5 mg/mL of heat-killed Mycobacterium tuberculosis H37 RA (non-viable) and 0.1 mg/mL of collagen. Injections were administered subcutaneously in 100 µL volume of the emulsion at the base of the tail. At 3 weeks after primary immunisation (14 weeks old), mice were administered a booster immunisation dose with bovine type II collagen in incomplete Freund’s adjuvant (Chondrex). The control groups (W and P) were injected with 100 µL PBS twice instead of the emulsion.
Arthritis was scored by a blinded examiner using a visual assessment scoring (VAS) system with a scale of 0–4 per limb, as described previously49 (link). Scoring was performed as follows: 0: no swelling or redness of paws or digits, 1: swelling and redness in one or two digits, 2: swelling and redness of the ankle or three or more digits or the midfoot, 3: swelling and redness of the ankle and midfoot or digits and midfoot, and 4: swelling and redness of the entire foot or ankylosis.
Arthritis was scored by a blinded examiner using a visual assessment scoring (VAS) system with a scale of 0–4 per limb, as described previously49 (link). Scoring was performed as follows: 0: no swelling or redness of paws or digits, 1: swelling and redness in one or two digits, 2: swelling and redness of the ankle or three or more digits or the midfoot, 3: swelling and redness of the ankle and midfoot or digits and midfoot, and 4: swelling and redness of the entire foot or ankylosis.
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