The effects of antiinflammatory and antiallergic drugs on cytokine release after stimulation of human whole blood by lipopolysaccharide and zymosan A.
IL-1 beta, IL-8, IL-6 and TNF alpha, derived from infiltrating leukocytes, are important mediators of inflammation in arthritic and allergic diseases. Heparinized human whole blood was evaluated as a model to study the effects of various classes of antiinflammatory drugs on cytokine release/biosynthesis from leukocytes. Whole blood was stimulated with zymosan A (1.5 mg/ml) or LPS (5 micrograms/ml) for 4 h to induce cytokine release. Dexamethasone was the most potent inhibitor of TNF alpha, IL-1 beta, IL-6 and IL-8 release from LPS stimulated blood leukocytes (IC50s of 0.19, 0.11 microM, 0.16 and 0.07 respectively). In LPS stimulated blood, SKF-86002, a 5-lipoxygenase/cytooxygenasae inhibitor, and rolipram, a PDE IV inhibitor, also inhibited the release of TNF alpha (IC50s of 33 and 11 microM, respectively), IL-1 beta (IC50s of 11 and 30 microM, respectively), IL-6 (IC50s of 56 and > 30, respectively) and IL-8 (IC50s of 6.7 and 15, respectively), whereas isoproterenol (1 microM) inhibited significantly only TNF alpha release. Nonsteroidal antiinflammatory drugs, 5-lipoxygenase inhibitors and immuno-suppressive drugs were inactive at 30 microM against LPS and zymosan A stimulation of cytokine release. Using zymosan A as the stimulus, only SKF-86002 (30 microM) showed significant inhibition of IL-1 beta (-59%). This 4 h human blood assay has the potential to identify novel inhibitors and sites of actions (e.g. transcription, post-transcriptional and secretion) of new antiinflammatory drugs.
Hartman
D A
DA
Wyeth-Ayerst Research, Princeton, NJ 08543, USA.
Ochalski
S J
SJ
Carlson
R P
RP
D018926:Anti-Allergic Agents / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007375:Interleukin-1 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000633:toxicity; D016859:Lipoxygenase Inhibitors; D008297:Male; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D012044:Regression Analysis; D020889:Rolipram; D013844:Thiazoles / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015054:Zymosan / Q000633:toxicity
D018926:Anti-Allergic Agents; D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D016207:Cytokines; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007375:Interleukin-1; D015850:Interleukin-6; D016209:Interleukin-8; D008070:Lipopolysaccharides; D016859:Lipoxygenase Inhibitors; D010726:Phosphodiesterase Inhibitors; D011760:Pyrrolidinones; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D015054:Zymosan; C052026:6-(4-fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo(2,1-b)thiazole; D020889:Rolipram