Difference in Legionella pneumophila growth permissiveness between J774.1 murine macrophage-like JA-4 cells and lipopolysaccharide (LPS)-resistant mutant cells, LPS1916, after stimulation with LPS.
To elucidate the role of the oxidative burst in macrophage resistance to Legionella infection, we examined a murine macrophage-like cell line, J774.1, for permissiveness to Legionella growth, using a mutant that has a selective defect in the oxidative burst after lipopolysaccharide (LPS) stimulation. Legionella pneumophila serogroup 1 was infected into J774.1 monolayers, and then the extent of bacterial growth was estimated by a CFU assay. Both the parental cell line, JA-4, and the LPS-resistant mutant, LPS1916, were permissive for Legionella growth but became nonpermissive after pretreatment with gamma interferon. However, pretreatment of LPS1916 cells with LPS failed to inhibit bacterial growth, although LPS-treated JA-4 cells exhibited inhibited multiplication of the bacteria. The bacterial growth inhibition in JA-4 and mutant LPS1916 cells was correlated with the extent of the oxidative burst in the cells, as judged by cytochrome c reduction but not nitrite production. Neither transferrin receptor expression nor the iron content in JA-4 and LPS1916 cells, with or without LPS treatment, was correlated with suppression of Legionella growth. These results suggest that the restriction of Legionella growth in J774.1 cells is due to a bactericidal effect of the oxidative burst rather than reduction of the iron supply to the intracellular bacteria and that the effectors are reactive oxygen intermediates and not reactive nitrogen intermediates.
Department of Bacteriology, National Institute of Health, Tokyo, Japan.
F
F
Kura
K
K
Suzuki
H
H
Watanabe
Y
Y
Akamatsu
F
F
Amano
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D007371:Interferon-gamma / Q000494:pharmacology; D007501:Iron / Q000032:analysis; D016952:Legionella pneumophila / Q000254:growth & development*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects* / Q000235:genetics; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D009154:Mutation; D011990:Receptors, Transferrin / Q000302:isolation & purification; D016897:Respiratory Burst; D019323:omega-N-Methylarginine
D008070:Lipopolysaccharides; D011990:Receptors, Transferrin; D019323:omega-N-Methylarginine; D007371:Interferon-gamma; D001120:Arginine; D007501:Iron