Stimulated murine peritoneal macrophages in suspension and cell culture: Enzyme determinations by biochemical and histochemical means

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Abstract
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.