Isolation of Salmonella wien heat-labile enterotoxin

Abstract

An enterotoxic activity has been identified in culture filtrates of Salmonella wien. The enterotoxin causes fluid accumulation in rabbit ligated ileal loops, firm induration and erythema in rabbit skin and morphological alteration in chinese hamster ovary (CHO) cell cultures; it was revealed by treatment with calcium phosphate gel, and purified on DEAE-Sephadex A-50 and BioGel A-1.5 m. The enterotoxic activity was eluted from the BioGel column in two peaks. Approximately 50-70% of the enterotoxic activity of the first peak, corresponding to the excluded volume, was resistant to heating at 75 degrees C for 30 min, while the activity of the second peak was completely destroyed by this treatment. From the heat-labile peak a protein, in homogeneous form, was isolated exploiting its affinity towards agarose gel filtration media. This protein, with enterotoxic activity was also present as shown by SDS-PAGE, in the first peak, eluted from the Bio-Gel column, where it appears to be closely associated with cell wall or membrane components and thus protected from heat denaturation. The isolated enterotoxin is stable in alkaline conditions but it is sensitive to acidic pH values; moreover, it stimulates adenylate cyclase in cell culture systems. Thus, it appears to possess properties similar to both cholera toxin and the heat-labile enterotoxin of Escherichia coli. These results indicate that the enterotoxin is a protein in nature and it is postulated that it may participate in the pathogenesis of S. wien infection.