This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Kennett, C.N. Articles by Eley, B.M. Search for Related Content PubMed PubMed Citation Articles by Kennett, C.N. Articles by Eley, B.M. Social Bookmarking                         What's this?

Localization of Active and Inactive Elastase, Alpha-1-Proteinase Inhibitor, and Alpha-2-Macroglobulin in Human Gingiva

Department of Periodontology, King's College School of Medicine and Dentistry, London, SE5 9RW, United Kingdom

S.W. Cox

Department of Periodontology, King's College School of Medicine and Dentistry, London, SE5 9RW, United Kingdom

B.M. Eley

Department of Periodontology, King's College School of Medicine and Dentistry, London, SE5 9RW, United Kingdom

Biochemically, there is usually much less elastase activity in gingival tissue than in crevicular fluid. The tissue distributions of active and inactive elastase and the endogenous inhibitors alpha-1-proteinase inhibitor ( ₁PI) and alpha-2-macroglobulin (₂M) were therefore compared. Inflamed tissue was obtained from chronic periodontitis patients, and cryostat sections were incubated with the histochemical elastase substrate MeOSuc-Ala-Ala-Pro-Val-MNA. Adjacent sections were examined immunocytochemically with antibodies to neutrophil elastase, ₁PI, ₂M, and leukocyte differentiation antigens. Antigenic elastase was widely distributed in CD15-positive granulocytes in both the epithelium and lamina propria as well as in granulomatous tissue from infrabony defects. However, there was very limited histochemical staining of these cells, and biochemical activity against the equivalent substrate MeOSuc-Ala-Ala-Pro-Val-AFC could be extracted only from sections with such staining. The pH optimum and effector response of the activity in the extracts were, nevertheless, consistent with those of leukocyte elastase. The large difference between the total elastase content of the tissue, as determined immunocytochemically, and the limited amount of active enzyme, as demonstrated histochemically, indicated that the majority was in an inactive form. The involvement of tissue inhibitors was suggested by the fact that extracts from sections with no histochemical staining reduced biochemical elastase activity in crevicular fluid. ₂M was found in many fibroblasts and also some CD68-positive macrophages, which additionally contained ₁PI. Both inhibitors appeared to be present extracellularly, since there was significant background staining. Thus, ₁PI and ₂M could contribute to the regulation of elastase activity in the gingiva.

Key Words: elastase • alpha-1-proteinase inhibitor • alpha-2-macroglobulin • gingiva • periodontitis.

REFERENCES

• Baggiolini M., Bretz U., Dewald B. (1978). Subcellular localization of granulocyte enzymes. In: Neutral proteases of human polymorphonuclear leukocytes. Havemann K, Janoff A, editors. Baltimore-Munich: Urban & Schwarzenberg, pp. 3-17.
• Condacci I., Cimasoni G., Ahmad-Zadeh C. (1982). 2macroglobulin in sulci from healthy and inflamed human gingivae. Infect Immun 36:66-71.[Abstract/Free Full Text]
• Condacci I., Cimasoni G., Rey M., Baehni P. (1988). In vitro synthesis of 2-macroglobulin by human gingival fibroblasts. Arch Oral Biol 33:407-412.[Medline] [Order article via Infotrieve]
• Cootauco CJ, Rauschenberger CR, Nauman RK (1993). Immunocytochemical distribution of human PMN elastase and cathepsin-G in dental pulp. J Dent Res 72:1485-1490.
• Cox SW, Eley BM ( 1987). Preliminary studies on cysteine and serine proteinase activities in inflamed human gingiva using different 7-amino-4-trifluoromethyl coumarin substrates and protease inhibitors. Arch Oral Biol 32:599-605.[Medline] [Order article via Infotrieve]
• Cox SW, Eley BM ( 1989). Detection of cathepsin B- and L-, elastase-, tryptase-, trypsin-, and dipeptidyl peptidase IV-like activities in crevicular fluid from gingivitis and periodontitis patients with peptidyl derivatives of 7-amino-4-trifluoromethyl coumarin. J Periodont Res 24:353-361.[CrossRef][Medline] [Order article via Infotrieve]
• Cox SW, Eley BM ( 1992). Cathepsin B/L-, elastase-, tryptase-, trypsin- and dipeptidyl peptidase IV-like activities in gingival crevicular fluid: a comparison of levels before and after basic periodontal treatment of chronic periodontitis patients. J Clin Periodontol 19:333-339.[CrossRef][Medline] [Order article via Infotrieve]
• Crocker J., Jenkins R., Burnett D. (1984). Immunohistochemical demonstration of leukocyte elastase in human tissues. J Clin Pathol 37:1114-1118.[Abstract/Free Full Text]
• Eley BM, Cox SW (1990). A biochemical study of serine proteinase activities at local gingival sites in human chronic periodontitis. Arch Oral Biol 35:23-27.[Medline] [Order article via Infotrieve]
• Eley BM, Cox SW (1992a). Cathepsin B/L-, elastase-, tryptase-, trypsin- and dipeptidyl peptidase IV-like activities in crevicular fluid: Correlation with clinical parameters in untreated periodontitis patients. J Periodont Res 27:62-69.[CrossRef][Medline] [Order article via Infotrieve]
• Eley BM, Cox SW (1992b) Correlation of gingival crevicular fluid proteases with clinical and radiological measurements of periodontal attachment loss. J Dent 20:90-99.[Medline] [Order article via Infotrieve]
• Eley BM, Cox SW (1992c). Cathepsin B/L-, elastase-, tryptase-, trypsin- and dipeptidyl peptidase IV-like activities in gingival crevicular fluid: a comparison of levels before and after periodontal surgery in chronic periodontitis patients. J Periodontol 63:412-417.[Medline] [Order article via Infotrieve]
• Giannopoulou G., Andersen E., Demeurisse C., Cimasoni G. (1992). Neutrophil elastase and its inhibitors in human gingival crevicular fluid during experimental gingivitis. J Dent Res 71:359-363.
• Gustafsson A., Asman B., Bergstrom K., Soder P.-Ö (1992). Granulocyte elastase in gingival crevicular fluid. A possible discriminator between gingivitis and periodontitis. J Clin Periodontol 19:535-540.[CrossRef][Medline] [Order article via Infotrieve]
• Havemann K., Gramse M., Gassel WD (1983). Cytochemical determination of granulocyte elastase and chymotrypsin in human myeloid cells and its application in acquired deficiency states and diagnosis of myeloid leukemia. Klin Wochenschr 61:49-56.[Medline] [Order article via Infotrieve]
• Hovi T., Mosher D., Vaheri A. (1977). Cultured human monocytes synthesize and secrete alpha-2-macroglobulin. J Exp Med 145:1580-1589.[Abstract/Free Full Text]
• Huynh C., Roch-Arveiller M., Meyer J., Giround JP (1992). Gingival crevicular fluid of patients with gingivitis or periodontal disease: evaluation of elastase-1 proteinase inhibitor complex. J Clin Periodontol 19:187-192.[CrossRef][Medline] [Order article via Infotrieve]
• Isaacson P., Jones DB, Millward-Sadler GH, Judd MA, Payne S. (1981). Alpha-1-antitrypsin in human macrophages. J Clin Pathol 34:982-990.[Abstract/Free Full Text]
• Kennett CN, Cox SW, Eley BM, Osman Iar (1993). Comparative histochemical and biochemical studies of mast cell tryptase in human gingiva. J Periodontol 64:870-877.[Medline] [Order article via Infotrieve]
• Kennett CN, Cox SW, Eley BM ( 1994). Comparative histochemical, biochemical and immunocytochemical studies of cathepsin B in human gingiva. J Periodont Res 29:203-213.[Medline] [Order article via Infotrieve]
• Kruze D., Menninger H., Fehr K., Boni A. ( 1976). Purification and some properties of a neutral protease from human leukocyte granules and its comparison with pancreatic elastase. Biochim Biophys Acta 438:503-513.[Medline] [Order article via Infotrieve]
• Kunimatsu K., Ichimaru E., Kato I., Kato Y., Sonoda Y., Aoki Y., et al. (1990). Granulocyte medullasin levels in gingival crevicular fluid from chronic adult periodontitis patients and experimental gingivitis subjects. J Periodont Res 25:352-357.[CrossRef][Medline] [Order article via Infotrieve]
• Mason DY, Cramer EM, Masse JM, Crystal R., Bassot JM, Breton-Gorius J. (1991). Alpha-1-antitrypsin is present within the primary granules of human polymorphonuclear leukocytes. Am J Pathol 139:623-628.[Abstract]
• Miller DR, Lamster IB, Chasens AI (1984). Role of the polymorphonuclear leukocyte in periodontal health and disease. J Clin Periodontol 11:1-15.[CrossRef][Medline] [Order article via Infotrieve]
• Ohlsson K., Olsson I., Tynelius-Bratthall G. (1974). Neutrophil leukocyte collagenase, elastase and serum protease inhibitors in human gingival crevices. Acta Odontol Scand 32:51-59.[Medline] [Order article via Infotrieve]
• Ornstein L., Janoff A., Sweetman FR, Ansley HR (1973). Histochemical demonstration of an elastase-like human neutrophil esterase. J Histochem Cytochem 21:411.
• Ponder BA, Wilkinson MM (1981). Inhibition of endogenous tissue alkaline phosphatase with the use of alkaline phosphatase conjugates in immunohistochemistry. J Histochem Cytochem 29:981-984.[Abstract]
• Rudolphus A., Stolk J., van Twisk C., van Noorden Cjf, Dijkman JH, Kramps JA (1992). Detection of extracellular neutrophil elastase in hamster lungs after intratracheal instillation of E. coli lipopolysaccharide using a fluorogenic, elastase-specific, synthetic substrate. Am J Pathol 141:153-160.[Abstract]
• Schroeder HE (1977). Histopathology of the gingival sulcus. In: The borderland between caries and periodontal disease. Lehner T, editor. London: Academic Press, pp. 43-78.
• Skaleric U., Zajsek P., Cvetko E., Lah T., Babnik J. (1986). Alpha 2-macroglobulin in gingival fluid: correlation with alveolar bone loss in periodontal disease. J Clin Periodontol 13:833-836.[Medline] [Order article via Infotrieve]
• Starkey PM (1977). Elastase and cathepsin G: the serine proteinases from human neutrophil leucocytes and spleen. In: Proteases in mammalian cells and tissues. Barrett AJ, editor. Amsterdam: Elsevier/North-Holland Biomedical Press, pp. 57-89.
• Starkey PM, Barrett AJ (1976). Human lysosomal elastase. Catalytic and immunological properties. Biochem J 155:265-271.[Medline] [Order article via Infotrieve]
• Travis J. (1988). Structure, function, and control of neutrophil proteinases. Am J Med 84:37-42.[Medline] [Order article via Infotrieve]
• Travis J., Salvesen GS (1983). Human plasma proteinase inhibitors. Ann Rev Biochem 52:655-709.[CrossRef][Medline] [Order article via Infotrieve]
• Zafiropoulos GGK, Flores-de-Jacoby L., Todt G., Kolb G., Havemann K., Tatakis DN (1991). Gingival crevicular fluid elastase inhibitor complex: correlation with clinical indices and subgingival flora. J Periodont Res 26:24-32.[CrossRef][Medline] [Order article via Infotrieve]

Journal of Dental Research, Vol. 74, No. 2, 667-674 (1995)
DOI: 10.1177/00220345950740020701


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Kennett, C.N. Articles by Eley, B.M. Search for Related Content PubMed PubMed Citation Articles by Kennett, C.N. Articles by Eley, B.M. Social Bookmarking                         What's this?