The degradation of basement membrane is important for leukocyte migration, tissue remodeling, wound healing, angiogenesis and tumor invasion. Type IV collagen, the major component of basement membrane, is degraded by type IV collagenase (CIVase) which usually exists in a latent form. The latent CIVase can be activated in vivo to an active form whose activity can be regulated by endogenous tissue inhibitor of metalloproteinase type II (TIMP2). As yet there is no specific biological, enzymatic or antibody based diagnostic kit commercially available with acceptable sensitivity for assaying CIVase or TIMP2. We have recently generated monoclonal antibodies (mAbs) against CIVase. These mAbs have been characterized by immunoprecipitation, Western blot, and immunocytochemical labeling of CIVase. We will use a similar approach to generate anti-TIMP2 mAbs and use these mAbs to develop quantitative enzyme-linked immunosorbent assay (ELISA), immunohistochemical and flow cytometry test for examining CIVase and TIMP2 level in the human body fluids and tissues. In phase II, we will use these mAbs to evaluate clinical correlations between CIVase and prognosis levels of patients with diseases involving the breakdown of basement membrane. We will also isolate cDNA coding for these anti- CIVase and anti-TIMP2 mAbs that show neutralization activity to generate single chain antigen-binding proteins or humanized chimeric antibodies. These molecules will be tested for potential therapeutic uses in wound healing, angiogenesis, treatment of rheumatoid arthritis and prevention of cancer metastasis.
| Hoyhtya, M; Fridman, R; Komarek, D et al. (1994) Immunohistochemical localization of matrix metalloproteinase 2 and its specific inhibitor TIMP-2 in neoplastic tissues with monoclonal antibodies. Int J Cancer 56:500-5 |
