Cytotoxic effect of the culture supernatant of Clostridium tetani.

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Abstract
The cytotoxic effect of the culture supernatant of C. tetani on mouse leukemia L1210 cells was studied. The cytotoxicity of the culture supernatant reached a maximum at 24 h after the beginning of cultivation. The supernatant fluid was fractionated by addition of ammonium sulfate and the fraction that showed the highest specific cytotoxic activity was subjected to gel filtration. The cytotoxic activity was separated in 2 peaks in parallel with the hemolytic activity; both activities were decreased by preincubation with cholesterol. An independently purified fraction of tetanolysin also exhibited cytotoxic activity. One of the cytotoxic substances was tetanolysin, a hemolysin produced by C. tetani. Tetanolysin potentiated the cytotoxicity of 5-fluorouracil in combined treatment.