Reduction of no synthase expression and tumor necrosis factor alpha production in macrophages by amphotericin B lipid carriers.
The present study compared the abilities of different lipid carriers of amphotericin B (AMB) to activate murine peritoneal macrophages, as assessed by their capacities to produce nitric oxide (NO) and tumor necrosis factor alpha (TNF-alpha). Although AMB alone did not induce NO production, synergy was observed with gamma interferon but not with lipopolysaccharide. This synergy could not be explained by the mobilization of the nuclear activation factor NF-kappaB by AMB. On the other hand, AMB induced TNF-alpha production without a costimulator and no synergy was observed. Anti-TNF-alpha antibodies did not influence NO production, and an inhibitor of NO synthase did not affect TNF-alpha production, indicating that the production of one of these effector molecules was independent of that of the other. The incorporation of AMB into lipid carriers reduced NO and TNF-alpha production with all formulations but more so with liposomes than with lipid complexes. NO production was correlated with the induction of NO synthase II, revealed by Western blotting. The extent of association of AMB with macrophages depended on the formulation, especially on the AMB/lipids ratio: the higher the ratio was, the greater the AMB association with macrophages. However, there was no clear correlation between AMB association with macrophages, whether internalized or bound to the membrane, and immunostimulating effects. These results may explain the reduced toxicities of lipid-based formulations of AMB.
Larabi
M
M
Laboratoire de Physico-Chimie-Pharmacotechnie-Biopharmacie, UMR CNRS 8612, France.
Legrand
P
P
Appel
M
M
Gil
S
S
Lepoivre
M
M
Devissaguet
J
J
Puisieux
F
F
Barratt
G
G
D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology*; D000666:Amphotericin B / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D000935:Antifungal Agents / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004337:Drug Carriers; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008081:Liposomes; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D009573:Nitrites / Q000378:metabolism; D014176:Protein Biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D000276:Adjuvants, Immunologic; D000935:Antifungal Agents; D004337:Drug Carriers; D008081:Liposomes; D016328:NF-kappa B; D009573:Nitrites; D014409:Tumor Necrosis Factor-alpha; D000666:Amphotericin B; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase