Comparison of the PK(15)- and WEHI 164 (clone 13)-based bioassays for detection of porcine tumor necrosis factor.
OBJECTIVE
To determine relative sensitivities of the PK(15)- and WEHI 164(13)-based bioassays for detection of tumor necrosis factor alpha (TNF).


METHODS
Recombinant human, murine, and porcine INF, and serum from pigs given endotoxin IV.


METHODS
Two cell lines were used as targets for recombinant human, murine, and porcine TNF cytotoxicity bioassays. Pigs were given sublethal doses of endotoxin to obtain serum samples containing high activity of porcine TNF. Serum TNF activity was tested, using both cell lines. Viable cells were detected by addition of dimethylthiazol diphenyltetrazolium bromide after 18 to 20 hours' incubation with samples containing TNF.


RESULTS
The 2 cell lines tested had different sensitivities to human, murine, and porcine TNF. Compared with WEHI 164(13) cells, PK(15) cells were 50 times less sensitive to murine TNF and 15 times less sensitive to human TNF. However, PK(15) cells were 4 times more sensitive to recombinant porcine TNF and 15 times more sensitive to porcine serum containing TNF.


CONCLUSIONS
The PK(15) cell line was more sensitive to porcine TNF-mediated lysis than was the WEHI 164(13) cell line. The PK(15)-based TNF bioassay will be especially useful for study of infectious disease processes in swine, particularly where low activity of TNF exists.
Boury
N M
NM
Department of Veterinary Physiology and Pharmacology, Iowa State University, Ames 50011, USA.
Stabel
T J
TJ
Kehrli
M
M
Taylor
M
M
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D001427:Bacterial Toxins / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods / Q000662:veterinary*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004731:Endotoxins / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007118:Immunoassay / Q000379:methods / Q000662:veterinary; D007668:Kidney / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D011994:Recombinant Proteins / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013552:Swine / Q000097:blood*; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism / Q000494:pharmacology*
D000906:Antibodies; D001427:Bacterial Toxins; D004731:Endotoxins; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; C027952:salmonella toxin; C022616:thiazolyl blue