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An assessment of the in vitro toxicology of Clostri di urn perfringens type D -toxin in human and animal cells

R W Titball

C D Lindsay

Defence Evaluation and Research Agency, CBD Porton Down, Salisbury, Wiltshire SP4 OJQ, UK

The epithelial Madin Darby canine kidney (MDCK) cell line and 17 human cell lines were examined for sensitivity to Clostridium perfringens type D a-toxin. MDCK cells were confirmed as being sensitive to the toxin. In addition, the Caucasian renal leiomyoblastoma (G-402) human cell line was identified as being -toxin sensitive. Using the MTS/PMS assay system the concentration of toxin reducing cell culture viability by 50% (LC₅₀) was found to be 2 ug/ml in MDCK cells. The LC₅₀ for G-402 cells was 280,ug/ml. a-Toxin was found to be rapid acting in MDCK cells exposed to a maximum lethal dose of the toxin (40% loss of viability after a 0.5 h exposure), but slower acting in G-402 cells (40% loss of viability after 1.7 h exposure). Photomicrography of toxin exposed cultures indicated necrotic cell death on exposure to a-toxin. Investigations using an antibody probe indicated that -toxin could bind to many cell surface proteins in both MDCK, G-402 and a toxin insensitive human cell line (CAKI-2). It has previously been found that the toxin may bind to the cell surface via glycosylated moieties. However, exposing MDCK and G-402 cells to a-toxin in the presence of sialic acid and several different sugars did not reduce the lethal effects of the toxin.

Key Words: Clostridium perfringens • a-toxin; MDCK • G-402 • MT

Human & Experimental Toxicology, Vol. 19, No. 2, 108-116 (2000)
DOI: 10.1191/096032700678815710


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