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Bacterial Lipopolysaccharide Induces Release of Tumor Necrosis Factor‐α From Bovine Peripheral Blood Monocytes and Alveolar Macrophages In Vitro
Author(s) -
Adams Jeffrey L.,
Czuprynski Charles J.
Publication year - 1990
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.48.6.549
Subject(s) - lipopolysaccharide , tumor necrosis factor alpha , monocyte , cytotoxicity , in vitro , biology , microbiology and biotechnology , fibrosarcoma , monoclonal antibody , macrophage , cytotoxic t cell , recombinant dna , immunology , antibody , biochemistry , genetics , gene
Abstract In this study, we demonstrate that freshly adherent bovine monocytes release tumor necrosis factor‐α (TNF‐α) in response to stimulation with bacterial lipopolysaccharide (LPS). TNF‐α was detected using actinomycin D‐treated WEHI‐164 murine fibrosarcoma cells as targets in an 18 hr cytotoxicity assay. Doses of LPS from 20 ng/ml to 20 μg/ml were capable of inducing bovine TNF‐α. The kinetics of TNF‐α release from bovine monocytes demonstrated peak levels of cytotoxic activity at 1–3 hr post‐LPS treatment, with a subsequent decline to background levels by 18 hr post‐LPS treatment. A monoclonal antibody that neutralizes recombinant human TNF‐α (rHuTNF‐α) significantly reduced the cytotoxicity of LPS‐stimulated bovine monocyte culture supematants. Size exclusion high‐performance liquid chromatography (HPLC) analysis of LPS‐stimulated monocyte and alveolar macrophage culture supematants resulted in a molecular weight elution profile similar to that of recombinant human TNF‐α. These elution profiles are consistent with the presence of multimers of TNF‐α. This is believed to be the first report of the in vitro production of bovine TNF‐α.