The influence of some metabolic inhibitors on in vitro phagocytizing macrophages

In the present work the uptake of foreign materials by macrophages has been studied in order to elucidate its possible energy-dependent mechanisms. We used monolayer cultures of macrophages from human peripheral venous blood, treated with the following metabolic inhibitors: iodoacetic acid, fluoroacetic acid, sodium fluoride, sodium malonate, sodium azide, 2–4-dinitrophenol, cycloheximide, and ouabain. The test assay was performed by using a zymosan particles suspension in Mc Coy 5 A medium supplemented as follows.The quantitation of phagocytosis was obtained by direct count of intracellular zymosan particles by oil 100X microscopy and the results were submitted to a statistical evaluation. The most effective inhibitor we found was iodoacetate, an inhibitor of anaerobic glycolysis, but fluoride, which acts on the same metabolic pathway at a different site, was quite ineffective. The same ineffectiveness we found for fluoracetate and malonate which act on the Krebs cycle. On the contrary, dinitrophenol (uncoupler of oxidative phosphorylation), azide (inhibitor of cytochrome linked-phosphorylation), ouabain (inhibitor of membrane ATPase activity) and cycloheximide (inhibitor of protein synthesis) give a remarkable decrease of index of phagocytosis after a 3h incubation. In conclusion, we can suppose that the energy-dependent phagocytosis is first depending on transport across the cell membrane (ATPase activity and protein synthesis) and second both on anaerobic glycolysis and oxidative phosphorylation.