Priming and stimulation of bovine neutrophils by recombinant human interleukin-1 alpha and tumor necrosis factor alpha.
We have examined the in vitro effects of two recombinant human monokines, interleukin-1 alpha (rHuIL-1 alpha) and tumor necrosis factor alpha (rHuTNF alpha), on bovine neutrophil functions. Both rHuIL-1 alpha (10 to 1,000 ng/ml) and rHuTNF alpha (5 to 50 ng/ml) directly stimulated the oxidative burst of bovine neutrophils as measured by Luminol-dependent chemiluminescence, superoxide anion generation, and hydrogen peroxide production. In addition, both rHuIL-1 alpha (1 to 1,000 ng/ml) and rHuTNF alpha (0.5 to 50 ng/ml) primed bovine neutrophils for an enhanced oxidative burst to subsequent stimulation with opsonized zymosan. Neutrophils pre-treated with either monokine exhibited an earlier, as well as stronger, zymosan-stimulated Luminol-dependent chemiluminescence response, as compared to untreated neutrophils. Exposure of bovine neutrophils to combinations of suboptimal doses of rHuIL-1 alpha (10 and 100 ng/ml) and rHuTNF alpha (0.5 and 5 ng/ml) resulted in a synergistic stimulation of Luminol-dependent chemiluminescence, whereas, no synergism was observed when using optimal doses of each monokine. Pre-incubation of bovine neutrophils with an optimal concentration of recombinant bovine interferon gamma (100 U/ml), and either rHuIL-1 alpha or rHuTNF alpha, further augmented the maximal oxidative response of neutrophils stimulated with opsonized zymosan. Bovine neutrophils released both primary and secondary granules in response to rHuIL-1 alpha and rHuTNF alpha, and also exhibited enhanced adherence in the presence of either monokine.
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A K
AK
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison 53706.
Czuprynski
C J
CJ
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000648:ultrastructure; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000494:pharmacology*; D007700:Kinetics; D008163:Luminescent Measurements; D008165:Luminol; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013481:Superoxides / Q000097:blood; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D007375:Interleukin-1; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D013481:Superoxides; D008165:Luminol