The modulation of IL-6 and TNF-alpha release by nitric oxide following stimulation of J774 cells with LPS and IFN-gamma.
Lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) induced nitric oxide synthase (NOS) activity, tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and prostaglandin (PG)E2 release in J774 cells, a murine macrophage cell line. The role of endogenous NO in modulating TNF-alpha and IL-6 release was investigated using N-iminoethyl-L-ornithine (L-NIO), a specific inhibitor of NOS. L-NIO (10-1000 microM) produced a concentration-dependent potentiation of LPS and IFN-gamma induced IL-6 release. Time-course studies demonstrated a significant potentiation of IL-6 release at 12 h with a maximum effect at 48 h. By contrast to its effects on IL-6, L-NIO significantly attenuated TNF-alpha release, and at 48 h reduced PGE2 release. The NO-donor S-nitroso-N-acetyl-penicillamine (SNAP,300 microM), significantly inhibited LPS and IFN-gamma induced IL-6 release, but potentiated TNF-alpha release. In addition, SNAP prevented the potentiation of IL-6 and the inhibition of TNF-alpha release by L-NIO. Stimulation of J774 cells with a combination of LPS and IFN-gamma for 24 h or 48 h reduced cell viability which was prevented by L-NIO. Furthermore, SNAP also reduced cell viability determined after 24 h incubation. These results indicate that NO can differentially modulate LPS and IFN-gamma-induced cytokine release from J774 cells, up-regulating TNF-alpha but down-regulating IL-6, and that NO is cytotoxic to these cells.
Deakin
A M
AM
Biology Division, Wellcome Research Laboratories, Beckenham, Kent.
Payne
A N
AN
Whittle
B J
BJ
Moncada
S
S
D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D007371:Interferon-gamma / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation; D051379:Mice; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D026423:S-Nitroso-N-Acetylpenicillamine; D013268:Stimulation, Chemical; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D015850:Interleukin-6; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; C065027:N(G)-iminoethylornithine; D026423:S-Nitroso-N-Acetylpenicillamine; D007371:Interferon-gamma; D009952:Ornithine; D019001:Nitric Oxide Synthase; D010396:Penicillamine