Inflammatory processes are characterized by the influx of inflammatory cells such as neutrophils and macrophages. My original proposal sought to better define the participation of PMNL in the connective tissue alterations seen during inflammation. Specifically, the objectives included: 1) purification and characterization of the collagenolytic metalloproteinases from human neutrophils; 2) characterization and regulation of the release of these proteinases; and 3) examination of the effect of neutrophil products on fibroblast collagen and collagenase production. Both neutrophil collagenase and gelatinase have been purified and characterized both biochemically and immunologically. Specific antibody probes for these proteinases have been developed. Release of the collagenolytic enzymes from neutrophils has been shown to be a sequential process and the modulation of this enzyme release appears to be significantly different than that previously reported for the fibroblast and synovial systems. While our studies of the neutrophil proteinases has permitted us to identify the similarities and differences between neutrophil enzymes and those from fibroblasts and synovial cells, little is known about the production of similar proteinases by macrophages, another common component of the inflammatory response. During the extension period, the interstitial collagenase and gelatinase from macrophages will be purified by a combination of standard chromatographic and immunoaffinity techniques and will be characterized by their molecular composition, substrate specificity, and inhibition by chemical and naturally occurring inhibitors. Monospecific polyclonal antisera will be prepared against each enzyme. Macrophage and neutrophil proteinases will be compared by structural analysis such as peptide mapping, amino acid composition and N-terminal amino acid sequence analysis and immunological studies utilizing immunoblot and ELISA techniques. These studies should permit the better definition of the proteinases which may play a role in the connective tissue remodelling seen in inflammatory processes as well as provide well defined probes for the study of these proteinases in vivo.
| Hibbs, M S (1992) Expression of 92 kDa phagocyte gelatinase by inflammatory and connective tissue cells. Matrix Suppl 1:51-7 |
| Hibbs, M S; Bainton, D F (1989) Human neutrophil gelatinase is a component of specific granules. J Clin Invest 84:1395-402 |
