Cytotoxic effect of T-2 mycotoxin on cells in culture as determined by a rapid colorimetric bioassay.
We developed a colorimetric assay for determining metabolic activity (viability) of cells exposed to toxic agents. This system is based on the ability of mitochondrial enzymes in viable cells to modify a tetrazolium salt into a blue formazan product that can be detected spectrophotometrically at 570 nm. The assay works equally well for mammalian and insect cell lines and at 48 hr color formation is linear over a cell input range of 1.56-50 X 10(4) cells/ml. The inhibitory effects of T-2 mycotoxin on tetrazolium cleavage in L929 cells is comparable to that observed for protein and DNA synthesis (50% inhibition = 6-8 ng/ml). Using this system to analyze the lethal effect of T-2 toxin on cells from various animal species, it was found that bovine cells were the most sensitive (50% inhibition at 2.2 ng/ml) while hamster cells were the most resistant (50% inhibition at 26.2 ng/ml). Murine cells exhibited intermediate sensitivity (50% inhibition at 10.9 ng/ml). Variable toxin susceptibility was also observed among different cell types. Lymphocytes were 3-fold more sensitive to the T-2 inhibitory effects than comparable tissue culture cell lines. These data indicate that the colorimetric assay system could have broad applications in toxicological studies. Further, the observed differences in species sensitivity may provide insight into the primary mechanism of the T-2 toxin-cell interaction that ultimately leads to cell death.
Holt
P S
PS
Veterinary Toxicology Research Unit, United States Department of Agriculture, College Station, Texas 77841.
Buckley
S
S
Norman
J O
JO
DeLoach
J R
JR
D000818:Animals; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D003124:Colorimetry; D006801:Humans; D007313:Insecta; D008214:Lymphocytes / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D012717:Sesquiterpenes / Q000633:toxicity*; D013045:Species Specificity; D013605:T-2 Toxin / Q000633:toxicity*; D013778:Tetrazolium Salts; D013844:Thiazoles
D012717:Sesquiterpenes; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue; D013605:T-2 Toxin