Soluble amoebicidal factors mediate cytolysis of Naegleria fowleri by activated macrophages.
Murine peritoneal macrophages activated in vivo with Corynebacterium parvum or bacille Calmette-Guérin, in contrast to resident macrophages, demonstrated significant cytolysis of the amoeba, Naegleria fowleri. Catalase and superoxide dismutase, both alone and in combination, failed to inhibit cytolysis of amoebae. N. fowleri amoebae demonstrated significant resistance to exogenously added hydrogen peroxide. The hydroxyl radical scavengers mannitol, thiourea, and dimethyl sulfoxide, as well as anaerobic conditions, failed to inhibit the amoebicidal activity of activated macrophages. Actinomycin D, cycloheximide, and puromycin blocked macrophage amoebicidal activity. Conditioned medium (CM) from lipopolysaccharide-stimulated, but not unstimulated, cultures of activated macrophages was capable of mediating cytolysis of N. fowleri amoebae. Cytolytic activity was recovered by ammonium sulfate precipitation of CM. Heat treatment of the CM inactivated cytolytic activity. Results indicate soluble proteins of activated macrophage origin to be responsible for the amoebicidal activity.
D000656:Amoeba / Q000276:immunology*; D000693:Anaerobiosis; D000818:Animals; D003470:Culture Media; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D003603:Cytotoxins / Q000032:analysis / Q000502:physiology*; D005260:Female; D005590:Fractional Precipitation; D005609:Free Radicals; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000494:pharmacology; D006878:Hydroxides / Q000037:antagonists & inhibitors; D017665:Hydroxyl Radical; D008262:Macrophage Activation / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D014176:Protein Biosynthesis; D012313:RNA / Q000096:biosynthesis; D013481:Superoxides / Q000037:antagonists & inhibitors
D003470:Culture Media; D003603:Cytotoxins; D005609:Free Radicals; D006878:Hydroxides; D013481:Superoxides; D017665:Hydroxyl Radical; D012313:RNA; D006861:Hydrogen Peroxide