Synergistic effect of coumarin (1,2 benzopyrone) and endotoxin in the induction of human interleukin-1.

Coumarin as well as its derivatives 7-OH coumarin and 4-OH coumarin were found to stimulate interleukin-1 beta (IL-1 beta) release from freshly isolated human mononuclear cells (MNC) if the culture medium contained fetal calf serum. Under serum-free conditions, almost no induction of IL-1 beta release was observed and the former effect could be completely eliminated by polymyxin B. Therefore, the combined action of endotoxin and coumarin was tested on MNC IL-1 beta production. The coumarins were able to potentiate human MNC IL-1 beta production by lipopolysaccharide (LPS) in a dose-dependent manner. That the effect was due to the presence of coumarins and not endotoxin contamination was shown by negative Limulus amebocyte lysate tests and pre-incubation of the coumarins with polymyxin B-agarose. The latter procedure was able to block endotoxin induced IL-1 beta production but the synergism between coumarin and endotoxin was not influenced by pre-incubating the coumarins with polymyxin B-agarose. Cycloheximide as well as actinomycin D eliminated the induction of IL-1 release by coumarin and LPS demonstrating that the cytokine was newly synthesized after MNC stimulation. In addition, both the total amount of MNC IL-1 beta (cell-associated + extracellular) and the extracellular portion of the cytokine were synergistically decreased if coumarin or its derivatives were added to endotoxin-stimulated cultures. Synergism of coumarin and endotoxin in the induction of interleukin-6 or tumour necrosis factor-alpha could be observed in a smaller percentage of donors. These findings demonstrate an immunomodulatory effect of coumarin on cytokine production by monocytes in vitro which might help to explain some of the biological activities attributed to the drug upon its application in tumour patients.