Charoenpornsook K, Fitzpatrick J L, Smith J E
Department of Bioscience and Biotechnology, University of Strathclyde, Glasgow, UK.
Mycopathologia. 1998;143(2):105-11. doi: 10.1023/a:1006971724678.
The effects of four mycotoxins, T-2 toxin, deoxynivalenol (DON), ochratoxin A (OTA) and fumonisin B1 (FB1) on the response of bovine peripheral blood mononuclear cells (PBM) in vitro to the mitogens concanavalin A (Con A), phytohaemagglutinin A (PHA) and pokeweed mitogen (PWM) were assayed after 4 days' incubation using 3H-thymidine uptake and the MTT bioassay. The concentrations of mycotoxin required to reduce the proliferative response of PBM by 50% for Con A, PHA and PWM as measured by 3H-thymidine incorporation was for T-2 toxin 0.30, 0.40 and 0.18 ng ml-1; for DON 0.07, 0.09 and 0.04 microgram ml-1; for OTA 0.10, 0.20 and 0.15 microgram ml-1, and for FB1 35, 18 and 11 micrograms ml-1 respectively. The concentrations of mycotoxin required to reduce the proliferative response of PBM by 50% for Con A, PHA and PWM as measured by the MTT bioassay were for T-2 toxin 2.0, 2.0 and 1.0 mg ml-1; for DON 0.70, 0.50 and 0.50 microgram ml-1; OTA 1.5, 1.5 and 1.5 micrograms ml-1; and FB1 > 50, > 50 and 20 micrograms ml-1 respectively. Further cytotoxicity assays including the LDH bioassay and Trypan blue exclusion were performed only on Con A-stimulated PBM cells after 72 h incubation. With the LDH-bioassay the 50% inhibition levels were T-2 toxin 0.3 ng ml-1, DON 0.4 microgram ml-1, OTA 1.4 micrograms ml-1 and FB1 3.5 micrograms ml-1; for Trypan blue uptake the 50% inhibition levels were T-2 toxin 5 ng ml-1, DON 2.3 micrograms ml-1 and OTA 4 micrograms ml-1 respectively.
使用³H-胸腺嘧啶核苷摄取法和MTT生物测定法,在孵育4天后,检测了四种霉菌毒素,即T-2毒素、脱氧雪腐镰刀菌烯醇(DON)、赭曲霉毒素A(OTA)和伏马菌素B1(FB1)对牛外周血单个核细胞(PBM)体外对促有丝分裂原刀豆球蛋白A(Con A)、植物血凝素A(PHA)和商陆有丝分裂原(PWM)反应的影响。通过³H-胸腺嘧啶核苷掺入法测定,使PBM对Con A、PHA和PWM的增殖反应降低50%所需的霉菌毒素浓度,对于T-2毒素分别为0.30、0.40和0.18 ng/ml;对于DON分别为0.07、0.09和0.04 μg/ml;对于OTA分别为0.10、0.20和0.15 μg/ml,对于FB1分别为35、18和11 μg/ml。通过MTT生物测定法测定,使PBM对Con A、PHA和PWM的增殖反应降低50%所需的霉菌毒素浓度,对于T-2毒素分别为2.0、2.0和1.0 mg/ml;对于DON分别为0.70、0.50和0.50 μg/ml;对于OTA分别为1.5、1.5和1.5 μg/ml;对于FB1分别>50、>50和20 μg/ml。仅在孵育72小时后,对Con A刺激的PBM细胞进行了包括LDH生物测定法和台盼蓝排斥试验在内的进一步细胞毒性试验。对于LDH生物测定法,50%抑制水平分别为:T-2毒素0.3 ng/ml、DON 0.4 μg/ml、OTA 1.4 μg/ml和FB1 3.5 μg/ml;对于台盼蓝摄取,50%抑制水平分别为:T-2毒素5 ng/ml、DON 2.3 μg/ml和OTA 4 μg/ml。