Production of a unique cytotoxin by Klebsiella oxytoca.

Certain strains of Klebsiella oxytoca isolated from patients with hemorrhagic enterocolitis produced a unique cytotoxin. The cytotoxin induced rounding of tissue culture cells, such as HEp-2, Vero, CHO and HeLa cells. The induced morphologic changes were indistinguishable between cell types. Seventy to 80% of the rounded cells died in 48 h incubation. The cytotoxin was purified 1000-fold from culture supernatant by Sephadex G-25 and Bio-Gel P-2 gel filtration followed by reversed-phase high-performance liquid chromatography. The molecular weight of the purified cytotoxin was estimated to be less than or equal to 651 by mass spectrometry. The minimum concentrations of the purified cytotoxin required to cause 50% of rounding of cells were 0.6 micrograms/ml for HEp-2, 0.8 micrograms/ml for Vero, 0.8 micrograms/ml for CHO and 1.4 micrograms/ml for HeLa cells. The type strain of K. oxytoca, ATCC 13182, did not produce the cytotoxin and only the clinically isolated strains did, suggesting that the cytotoxin may play a role in the pathogenesis of the organisms.