1. ¹Department of Pathology, School of Medicine, Keio University, Tokyo, Japan
2. ²Fuji Photo Film Co., Ltd., Institute of Medical Science, University of Tokyo, Tokyo, Japan
3. ³Department of Cancer Cell Research, Institute of Medical Science, University of Tokyo, Tokyo
4. ⁴Department of Orthopaedic Surgery, School of Medicine, Chiba University, Chiba, Japan
5. ⁵Biopharmaceutical Department, Fuji Chemical Industries, Ltd., Takaoka, Japan
6. ⁶Department of Virology and Oncology, Cancer Research Institute, Kanazawa University, Kanazawa, Japan

Correspondence: Dr. Y. Okada, Department of Pathology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-0016, Japan. Fax: 81 3 3353 3290; E-mail: okada@med.keio.ac.jp

Received 10 December 1999.

Abstract

In vitro, membrane-type matrix metalloproteinases (MT-MMP) are known to activate the zymogen of MMP-2 (proMMP-2, progelatinase A), which is one of the key MMP in joint destruction in rheumatoid arthritis. In the present study, we examined the production and activation of proMMP-2, and the expression of MT1-MMP, MT2-MMP, and MT3-MMP, their correlation with proMMP-2 activation, and their localization in rheumatoid synovial tissue. Using sandwich enzyme immunoassay and gelatin zymography techniques, proMMP-2 production levels and activation ratios were found to be significantly higher in rheumatoid synovium compared with normal synovium (p < 0.01). Quantitative RT-PCR analyses demonstrated that MT1-MMP and MT3-MMP were expressed in all rheumatoid synovial tissue (30 of 30 cases), but that the mean expression level of MT1-MMP was approximately 11-fold higher than MT3-MMP. Significant correlation was found between the mRNA expression level of MT1-MMP and the activation ratio of proMMP-2 (p < 0.01). In situ hybridization indicated that the hyperplastic lining cells of rheumatoid synovium expressed MT1-MMP. Immunohistochemistry demonstrated that MT1-MMP was co-localized with MMP-2 and with a tissue inhibitor of metalloproteinase-2, and was mainly located in the rheumatoid synovial lining cells. In situ zymography of rheumatoid synovium showed gelatinolytic activity, predominantly in the lining cell layer. This activity was blocked when incubated with BB94, a specific MMP inhibitor. These results demonstrate that MT1-MMP plays an important role in the activation of proMMP-2 in the rheumatoid synovial lining cell layer, and suggest that its activity may be involved in the cartilage destruction of rheumatoid arthritis.