Original Article

Immunology and Cell Biology (1994) 72, 406–414; doi:10.1038/icb.1994.60

Attenuation of adjuvant arthritis in rats by treatment with oxygen radical scavengers

L Santos1 and PG Tipping1

1Centre for Inflammatory Diseases, Monash University, Department of Medicine, Monash Medical Centre, Clayton, Victoria, Australia

Correspondence: Dr P. G. Tipping, Dept of Medicine, Monash Medical Centre, Clayton Rd, Clayton, Vic. 3168, Australia.

Received 6 October 1993; Accepted 24 February 1994.

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Abstract

The contribution of reactive oxygen species (ROS), in particular hydroxyl radical (OH dot), to joint inflammation was examined in rats developing adjuvant arthritis (AA) by treatment with ROS scavengers dimethylthiourea (DMTU) and DMSO. Adjuvant arthritis was induced in Sprague-Dawley (SD) rats by a single intradermal (i.d.) injection of Mycobacterium tuberculosis (MT) in oil on day 0. By day 14, all rats exhibited arthritis in the hindlimbs and the majority had involvement of the forelimbs. A marked inflammatory cell influx (75% neutrophils) was present in the synovial fluid. These cells, in vitro, spontaneously produced OHdot (0.96plusminus0.28 OHdot units/h per 105 cells). In contrast, spontaneous OHdot production by normal circulating leucocytes was absent (0.07plusminus0.03 OHdot units/h per 105 cells). Adjuvant-injected rats were treated with DMTU (500, 250 and 100 mg/kg). DMSO (330 and 165 mg/kg) or saline (disease control) once daily on days 8, 9 and 10 and twice daily on days 11, 12 and 13 postadjuvant injection. Both DMTU and DMSO significantly reduced the clinical evidence of arthritis (clinical scores: DMTU [500 mg/kg] = 0, P < 0.0001; DMSO [3.0 mL/kg] = 0.4plusminus0.3, P < 0.01, compared with disease control = 2.3plusminus0.3). Synovial fluid cell accumulation was also significantly reduced (DMTU [500 mg/kg] = 0.5 plusminus 0.1 times 105 cells/four joints, P < 0.0001; DMSO [3.0 mL/kg] 2.75 plusminus 1.9 times 105 cells/four joints, P < 0.01 compared with disease control = 11.76 plusminus 1.7 times 105 cells/four joints). Neither treatment inhibited cutaneous delayed type hypersensitivity (DTH) to the disease inducing antigen. Furthermore, DMTU (500 mg/kg) did not cause neutropenia nor inhibit peritoneal neutrophil accumulation in response to a chemotactic stimulus. This study demonstrates the attenuation of adjuvant arthritis by ROS scavengers and suggests a pivotal role for ROS, particularly OHdot, in the mediation of joint inflammation in this disease.

Keywords:

adjuvant arthritis, dimethylthiourea, dimethylsulfoxide, hydroxyl radical, joint inflammation, Mycobacterium tuberculosis, reactive oxygen species, Sprague-Dawley.

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