Improved bioassay for the detection of porcine tumor necrosis factor using a homologous cell line: PK(15).
Close similarities of various physiological parameters makes the pig one of the preferred animal models for the study of human diseases, especially those involving the cardiovascular system. Unfortunately, the use of pig models to study diseases such as viral hemorrhagic fevers and endotoxic shock syndrome have been hampered by the lack of the necessary immunological tools to measure important immunoregulatory cytokines such as tumor necrosis factor (TNF). Here we describe a TNF-bioassay which is based on the porcine kidney cell line PK(15). Compared to the widely used murine fibroblastoid cell line L929, the PK(15) cell line displays a 100-1000-fold higher sensitivity for porcine TNF-alpha, a higher sensitivity for human TNF-alpha, and a slightly lower sensitivity for murine TNF-alpha. Using a PK(15) bioassay we can detect recombinant TNF-alpha as well as cytotoxic activity in the supernatants of lipopolysaccharide (LPS)-activated porcine monocytes at high dilutions. This suggests that the sensitivity of the test should permit the detection of TNF in biological specimens such as pig serum.
Bertoni
G
G
Institute of Veterinary Virology, University of Bern, Switzerland.
Kuhnert
P
P
Peterhans
E
E
Pauli
U
U
D000818:Animals; D001681:Biological Assay; D002460:Cell Line; D066298:In Vitro Techniques; D007668:Kidney; D007739:L Cells; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D009000:Monocytes / Q000378:metabolism; D011994:Recombinant Proteins; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000096:biosynthesis
D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha