Summary
Peritoneal exudate cells of mice were studied up to 14 days after i.p. injection of thioglycollate broth medium by means of conventional enzyme determinations and quantitative histochemical measurements of individual cells. Cells from the peritoneal cavity were either investigated immediately after harvesting or after culturing periods of up to six days for enzymic activities of aminopeptidase, esterase, lactate dehydrogenase and β-galactosidase. A fairly good correlation exists between biochemical determinations of aminopeptidase and esterase activity and the mean of histochemical data. Following a sharp increase in the number of cells after stimulation, aminopeptidase, esterase and lactate dehydrogenase activities per cell were found to be increased. Moreover, the cells taken three or five days after stimulation synthesized large amounts of aminopeptidase and esterase as shown by culturing experiments. This capacity of the cells was subsequently lost in cells harvested seven day after stimulation but β-galactosidase increased and lactate dehydrogenase was more readily released into culture supernatants. The increase in aminopeptidase and esterase was dependent on protein synthesis since it was abolished by cycloheximide. Thioglycollate broth medium provokes immigration of cells into the peritoneal cavity where cells apparently differentiate by increasing their aminopeptidase and esterase concentrations, and by raising their intracellular catabolism rates, which leads eventually to the decay of the cells. The different enzymic phenotypes and the large heterogeneity at any time point after stimulation presumably also reflect different functional properties during the inflammatory process.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alexander, P. (1976) The functions of the macrophage in malignant disease.Ann. Rev. Med. 27, 207–24.
Cohn, Z. A. (1978) The activation of mononuclear phagocytes: fact, fancy, and future.J. Immunol. 121, 813–6.
Drath, D. B., &Karnovsky, M. L. (1975) Superoxide production by phagocytic leucocytes.J. exp. Med,141, 257–62.
Edelson, P. J. &Cohn, Z. A. (1976) 5-Nucleotidase activity of mouse peritoneal macrophages. I. Synthesis and degradation in resident and inflammatory populations.J. exp. Med. 144, 1581–95.
Edelson, P. J. &Erbs, C. (1978) Plasma membrane localization and metabolism of alkaline phosphodiesterase I in mouse peritoneal macrophages.J. exp. Med. 147, 77–86.
Erb, P. &Feldmann, M. (1975) Role of macrophages inin vitro induction of T-helper cells.Nature, Lond. 254, 352–4.
Feldmann, M. (1972) Cell interactions in the immune responsein vitro. II. The requirement for macrophages in lymphoid cell collaboration.J. exp. Med. 135, 1049–58.
Guilbault, G. G. &Kramer, D. N. (1964) Fluorometric determination of lipase, acylase, alpha-and gamma-chymotrypsin and inhibitors of these enzymes.Analyt. Chem. 36, 409–14.
Hogg, N. &Parish, C. R. (1980) Surface antigens of the murine cytostatic peritoneal macrophage.Immunology,41, 187–93.
Hohorst, H. J. (1957) Enzymatische Bestimmung vonl(+)-Milchsäure.Biochem. Z. 328, 509–21.
Karnovsky, M. L., Lazdins, J., Drath, D. &Harper, A. (1975a) Biochemical characteristics of activated macrophages.Ann. New York.Acad. Sci. 256, 266–74.
Karnovsky, M. L., Lazdins, J. &Simmons, S. R. (1975b) Metabolism of activated mononuclear phagocytes at rest and during phagocytosis. InMononuclear Phagocytes in Immunity, Infection and Pathology (Edited byVan Furth, R.), pp. 423–438. Oxford: Black well Scientific Publications.
Karnovsky, M. L. &Lazdins, J. K. (1978) Biochemical criteria for activated macrophages.J. Immunol. 121, 809–13.
Lee, K. C. (1980) On the origin and mode of action of functionally distinct macrophage subpopulations.Molec. Cell. Biochem. 30, 39–55.
Mackaness, G. B. (1969) The influence of immunologically committed lymphoid cells on macrophage activityin vivo.J. exp. Med. 129, 973–92.
Mackaness, G. B. (1970) Cellular immunity. InMononuclear Phagocytes (edited byVan Furth R.), pp. 461–475. Oxford: Blackwell Scientific Publications.
Meltzer, M. S., Ruco, L. P., Boraschi, D. &Nacy, C. A. (1979) Macrophage activation for tumor cytotoxicity: analysis of intermediary reactions.J. Reticuloendoth. Soc. 26, 403–15.
Mitchell, G. F. (1979) Effector cells, molecules and mechanisms in host-protective immunity to parasites.Immunology 38, 209–23.
Morahan, P. S., Edelson, P. J. &Gass, K. (1980) Changes in macrophage ectoenzymes associated with anti-tumour activity.J. Immunol. 125, 1312–7.
Mosier, D. E. (1967) A requirement for two cell types for antibody formationin vitro.Science 158, 1573–75.
Nathan, C. F. &Root, R. K. (1977) Hydrogen peroxide release from mouse peritoneal macrophages. Dependence on sequential activation and triggering.J. exp. Med. 146, 1648–62.
Raz, A., Fogler, W. E. &Fidler, I. J. (1979) The effects of experimental conditions on the expression ofin vitro-mediated tumor cytotoxicity mediated by murine macrophages.Cancer Immunol. Immunother. 7, 157–63.
Schnyder, J. &Baggiolini, M. (1978) Secretion of lysosomal hydrolases by stimulated and nonstimulated macrophages.J. exp. Med. 148, 435–50.
Sellinger, O. Z., Beaufay, H., Jacques, P., Doyen, A. &De Duve, C. (1960) Intracellular distribution and properties of β-N-acetylglucosaminidase and β-galactosidase in rat liver.Biochem. J. 74, 450–6.
Serio, C., Gandour, D. M. &Walker, W. S. (1979) Macrophage functional heterogeneity: evidence for different antibody-dependent effector cell activities and expression of Fc-receptors among macrophage subpopulations.J. Reticuloendoth. Soc. 25, 197–206.
Skudlarek, M. D. &Swank, R. T. (1979) Biosynthesis of two lysosomal enzymes in macrophages. Evidence for a precursor of β-galactosidase.J. biol. Chem. 254, 9939–42.
Tucker, S. B., Pierre, R. V. &Jordon R. E. (1977) Rapid identification on monocytes in a mixed mononuclear cell preparation.J. Immunol. Meth. 14, 267–9.
Unkeless, J. C., Gordon, S. &Reich, E. (1974) Secretion of plasminogen activator by stimulated macrophages. J. exp. Med.139, 834–50.
Van Furth, R., Hirsch, J. G. &Fedorko, M. E. (1970) Morphology and peroxidase cytochemistry of mouse promonocytes monocytes and macrophages.J. exp. Med. 132, 794–805.
Wachsmuth, E. D. (1975) Aminopeptidase as a marker for macrophage differentiation.Expl Cell Res. 96, 409–12.
Wachmuth, E. D. (1976) Quantitation of enzymes in tissue sections by estimation of hydrolytic activity and antigenic determinants.Acta histochem. Suppl.16, 221–31.
Wachsmuth, E. D. &Stoye, J. F. (1977a) Aminopeptidase on the surface of differentiating macrophages: induction and characterization of the enzyme.J. Reticuloendoth. Soc. 22, 469–83.
Wachsmuth, E. D. &Stoye, J. P. (1977b) Aminopeptidase on the surface of differentiating macrophages: concentration changes on individual cells in culture.J. Reticuloendoth. Soc. 22, 485–97.
Wachsmuth, E. D. &Staber, F. G. (1977) Changes in membrane-bound aminopeptidase on bone marrow-derived macrophages during their maturationin vitro.Expl Cell Res. 109, 269–76.
Werb, Z., &Gordon, S. (1975a) Secretion of a specific collagenase by stimulated macrophages.J. exp. Med. 142, 346–60.
Werb, Z. &Gordon, S. (1975b) Elastase section by stimulated macrophages.J. exp. Med. 142, 361–77.
Wiener, E. &Curelaru, Z. (1975) The intracellular distribution of cathepsins and other acid hydrolases in mouse peritoneal macrophages.J. Reticuloendoth. Soc. 17, 319–32.
Yam, L. T., Li, C. Y. &Crosby, W. H. (1971) Cytochemical identification of monocytes and granulocytes.Am. J. clin. Pathol. 55, 283–90.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wachsmuth, E.D., Wüst, B. Stimulated murine peritoneal macrophages in suspension and cell culture: Enzyme determinations by biochemical and histochemical means. Histochem J 14, 221–237 (1982). https://doi.org/10.1007/BF01041217
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01041217