Injections were started on the third day after arthritis induction and were performed three times a week. In a second set of experiments, D8, the anti-eotaxin-2 antibody showing best protective

results, was tested in a dose–response model. Adjuvant arthritis was induced according to the above-described protocol. Animals (six rats per each condition) were treated with D8 intraperitoneally at a dose of 20 µg, 100 µg or 1000 µg, starting on day 3, three times weekly (D8 prevention group). A separate set of animals (six per condition) were treated with identical doses after arthritis onset (D8 treatment group). In order to compare the anti-inflammatory effect of D8 with that of a traditional anti-inflammatory agent of known efficacy, one group was treated with intraperitoneal methotrexate MG-132 in vitro (MTX), 0·25 mg/kg, once weekly, starting on day 3 after arthritis induction (MTX prevention group). An additional group was treated with MTX, 0·25 mg/kg once weekly, in combination with D8, 100 µg intraperitoneally given three times a week, starting on day 3 (combined D8–MTX prevention group). A control group was treated with PBS throughout the experiment. Body weight in grams was measured every other day as an indicator of systemic inflammation. For evaluation of paw swelling, ankle and wrist diameter in mm (to one place after the decimal point) were recorded

selleck chemical three times a week. Each paw was scored on a scale of 0–4 for the degree of swelling, erythema

and deformity (maximum score 16 per animal) as follows: 0 = normal, 1 = slight erythema and/or swelling of the ankle or wrist, 2 = moderate erythema and/or swelling of ankle or wrist, 3 = severe erythema and/or swelling of ankle or wrist and 4 = complete erythema and swelling of toes or fingers and ankle or wrist and inability to bend the ankle or wrist. Finger and toe swelling was recorded according to their partial contribution: ankles, each toe scored 0·2; wrist, each finger scored 0·25; the sum of all joints was calculated. Whole animal mobility was scored between 0 and 4 according to the following definitions: 0 = normal, Y-27632 2HCl 1 = slightly impaired, 2 = major impairment, 3 = does not step on paw and 4 = no movement. Data were analysed using spss software version 16·01. Student’s t-test was performed to identify significant differences between experimental groups. Three or more group means were compared by one-way analysis of variance, with an assumed significance level of P < 0·05. In these experiments, treatment was given before the appearance of clinical arthritis (prevention group). Effect of treatment with anti-eotaxin-2 antibodies on arthritis score.  Treatment with anti-eotaxin-2 monoclonal antibodies caused a significant reduction in arthritic score severity, compared to rats treated with PBS. This protective effect was evident in all three antibodies tested (G7, G8 and D8).