Bioassay for interleukin-1, interleukin-6, and tumor necrosis factor-like activities in canine sera.
To measure interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-like activities in canine serum, bioassays were conducted using human melanoma A375S1, IL-6 dependent murine hybridoma MH60.BSF2, and WEHI 164 murine sarcoma subclone 28-4. Clinically normal adult beagles were experimentally induced endotoxic shock by an intravenous injection of lipopolysaccharide or local inflammation by an intramuscular injection of turpentine oil. IL-1-like activity was detected in sera from dogs with endotoxic shock. IL-6 and TNF-like activities were detected in sera from both dogs with endotoxic shock and local inflammation. IL-1-like activity in sera from the dogs with endotoxic shock declined after dilution with either medium or serum obtained before treatment (pre-serum), but the IL-1-like activity was maintained to a greater extent in samples diluted with pre-serum compared to those diluted with medium. TNF-like activity declined equally after dilution with either medium or pre-serum. On the other hand, IL-6-like activity was inhibited at low dilution. It was, therefore, necessary to dilute the serum samples to 1:180 from dogs with endotoxic shock or 1:60 from dogs with local inflammation, in order to minimize the effect of inhibitory factors on IL-6-like activity. IL-6-like activity was neutralized by monoclonal antibody against murine IL-6 receptors. TNF-like activity was neutralized by anti-mouse TNF alpha rabbit serum. However IL-1-like activity was not neutralized by either anti-mouse or anti-human IL-1 rabbit serum.
Yamashita
K
K
Department of Veterinary Surgery 1, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan.
Fujinaga
T
T
Hagio
M
M
Miyamoto
T
T
Izumisawa
Y
Y
Kotani
T
T
D000818:Animals; D001681:Biological Assay; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004285:Dogs / Q000097:blood* / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D007249:Inflammation; D007375:Interleukin-1 / Q000097:blood* / Q000494:pharmacology; D015850:Interleukin-6 / Q000097:blood* / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000633:toxicity; D008297:Male; D008545:Melanoma; D051379:Mice; D011994:Recombinant Proteins / Q000494:pharmacology; D012016:Reference Values; D012513:Sarcoma, Experimental; D012772:Shock, Septic / Q000097:blood* / Q000276:immunology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology; D014425:Turpentine / Q000633:toxicity
D007375:Interleukin-1; D015850:Interleukin-6; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D014425:Turpentine