Abstract
Cytotoxicity of T-2 toxin, HT-2 toxin, acetyl T-2, neosolaniol, and T-2 tetraol was compared between normal human fibroblasts and mutant I-cell human fibroblasts, which only produce 10 to 15% of lysosomal hydrolases present in normal fibroblasts. Both cleavage of 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and cell count by hemocytometer were used for evaluations. For all toxins, dose-related effects on both types of cultures were evident. Cytotoxicity of the above mycotoxins on both cell lines were similar, indicating that lysosomal enzymes were not involved in the toxicity of T-2 toxin and its congeners. An inhibitor of lysosomal cysteine proteases (E-64) did not alter the cytotoxicity of T-2 toxin. The decreasing order of toxicity was T-2 toxin, HT-2 toxin, neosolaniol, acetyl T-2 toxin, and T-2 tetraol in both cell lines. When normal human fibroblasts were loaded with the fluorescent dye Lucifer yellow CH (LY), a subsequent treatment of T-2 toxin did not disrupt lysosomal membranes. The uptake of LY was not affected by T-2 toxin, which indicated that T-2 toxin did not interfere with the endocytic pathway. Results indicate that T-2 toxin and its congeners do not exert their primary toxic effect through lysosomal enzymes, membranes, or via the endocytic pathway.
Similar content being viewed by others
References
Ueno Y, Nakajima M, Sakai K, Ishii K, Sato N, Shimada N (1973) Comparative toxicology of trichothecene mycotoxins: Inhibition of protein synthesis in animal cells. J Biochem 74:285–296
Thompson WL, Wannemacher, Jr RW (1986) Structure-function relationships of 12, 13-epoxytrichothecene mycotoxins in cell culture: comparison to whole animal lethality. Toxicon 24:985–994
Pace JG, Watts MR, Canterbury WJ (1988) T-2 mycotoxin inhibits mitochondrial protein synthesis. Toxicon 26:88–95
Bunner DL, Morris ER (1988) Alteration of multiple cell membrane functions in L-6 myoblasts by T-2 toxin: an important mechanism of action. Toxicol Appl Pharmacol 92:113–121
DeLoach JR, Andrews K, Naqi A (1987) Interaction of T-2 toxin with bovine carrier erythrocytes: effects on cell lysis, permeability, and entrapment. Toxicol Appl Pharmacol 88:123–131
DeLoach JR, Gyongyossy-Issa MIC, Khachatourians GG (1989) Species-specific hemolysis of erythrocytes by T-2 toxin. Toxicol Appl Pharmacol 97:107–112
Gyongyossy-Issa MIC, Khachatourians GG (1984) Interaction of T-2 toxin with murine lymphocytes. Biochim Biophys Acta 803:197–202
Schappert KT, Khachatourians GG (1984) Influence of the membrane on T-2 toxin toxicity inSaccharomyces spp. Appl Environ Microbiol 47:681–684
Hayes MA, Bellamy JCE, Schiefer HB (1980) Subacute toxicity of dietary T-2 toxin in mice: morphological and hematological effects. Can J comp Med 44:203–218
Hayes MA, Schiefer HB (1979) Quantitation and morphological aspects of cutaneous irritation by trichothecene mycotoxins. Fd Cosmet Toxicol 17:611–621
Joffe AZ (1986)Fusarium Species: Their Biology and Toxicology. Toronto, John Wiley and Sons. Toronto
Lusky I, Mor N (1981) Experimental alimentary toxic aleukia in cats. Laboratory Animal Sei 31:43–47
Pang VF, Lorenzana RM, Beasley VR, Buck WB, Haschek WM (1987) Experimental T-2 toxicosis in swine. III. Morphological changes following intravascular administration of T-2 toxin. Fund Appl Toxicol 8:298–309
Weaver GA, Kurtz HJ, Bates FY (1978) Acute and chronic toxicity of T-2 mycotoxin in swine. Vet Ree 103:531–535
Ueno Y (1983) Trichothecenes: Chemical, Biological, and Toxicological Aspects. Developments in Food Science. Elsevier, New York
Conrady-Lorck S, Gareis M, Feng XC, Amselgruber W, Forth W, Fichtl B (1988) Metabolism of T-2 toxin in vascularly autoperfused jejunal loops of rats. Toxicol Appl Pharmacol 94:23–33
Pace JG (1986) Metabolism and clearance of T-2 mycotoxin in perfused rat livers. Fund Appl Toxicol 7:424–433
PaceJG, Watts MR, Burrows EP, Dinterman RE, Matson C, Hauer EC, Wannemacher, Jr RW (1985) Fate and distribution of 3H-labeled T-2 mycotoxin in guinea-pigs. Toxicol Appl Pharmacol 80:377–385
Sintov A, Bialer M, Yagen B (1988) Pharmacokinetics and protein binding of trichothecene mycotoxins, T-2 and HT-2 toxin, in dogs. Toxicon 26:153–161
Johensen H, Odden E, Johnsen BA, Fonnum F (1988) Metabolism of T-2 toxin by blood cell carboxylesterases. Biochem Pharmacol 37:3193–3197
Trusal LR (1986) Metabolism of T-2 mycotoxin by cultured cells. Toxicon 24:597–693
Yoneyama M, Sharma RP (1986) Cytotoxicity of Fusarium T-2 in cultured Madin-Darby bovine (MDBK) and primary fetal bovine kidney (PFBK) cells. Mycotoxin Res 2:79–88
Neufeld EG, Lim TW, Shapiro LJ (1975) Inherited disorders of lysosomal metabolism. Ann Rev Biochem 44:357–376
Mosman T (1983) Rapid calorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Wilson PD, Firestone RA, Lenard J (1987) The role of lysosomal enzymes in killing of mammalian cells by the lysosomotropic detergent N-dodecylimidazole. J Cell Biol 104:1223–1229
Mutoh A, Ishii K, Ueno Y (1988) Effects of radioprotective compounds and anti-inflammatory agents on the acute toxicity of trichothecenes. Toxicol Lett 40:165–174
Guy ton AC (1986) Textbook Of Medical Physiology. WB Saunders Company, Philadelphia, PA
Adallh MA, Biellman JF (1980) Activity determination of 3-iodopyridine-adenine dinucleotide and its phosphate as hydride acceptors in the presence of dehydrogenases using a coupled redox system. Eur J Biochem 112:331–333
Stewart WW (1978) Functional connections between cells as revealed by dyecoupling with a highly fluorescent naphthalimide tracer. Cell 14:741- 759
Barrett AJ, Kembhavi AA, Brown MA, Kirschke H, Knight CG, Tamai M, Hanada K (1982) L-trans-epoxysuccinylleucylamido (4-guanidino) butane (E-64) and its analogues as inhibitors of cysteine proteinases including cathepsins B, H and L. Biochem J 201:189–198
Author information
Authors and Affiliations
Additional information
Published as Utah Agricultural Experiment Station, Utah State University Journal, Paper No 3901References
Rights and permissions
About this article
Cite this article
Kim, YW., Sharma, R.P. & Eisner, Y. Effects of T-2 toxin and its congeners on membrane functions of cultured human fibroblasts. Mycotox Res 7, 19–28 (1991). https://doi.org/10.1007/BF03192160
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03192160