[On nonspecific detoxification of bacterial toxins].
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The quality of tetanus toxin affected the sensitivity of the toxin neutralization (TN) test greatly. By using purified toxin a minimum level of 0.001 IU/ml of tetanus antitoxin could be detected whereas with crude toxin a level of 0.025 IU/ml only could be detected. The TN test described in this report permitted titration of tetanus antitoxin in twofold dilution steps from levels as low as 0.001 IU/ml using 0.6 ml of serum only at the L+/5000 level of purified tetanus toxin. Treatment of the sera with 2-mercaptoethanol (2-ME) did not affect the TN titres showing that the TN test detects the neutralizing antibodies (IgG) which are not affected by 2-ME. The TN test was found to be a highly sensitive and reproducible test.
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A completely synthetically produced peptide vaccine, consisting of the 18-aminoacid Escherichia coli heat-stable toxin and the 26-aminoacid epitope of the heat-labile toxin B subunit, was given orally to thirteen volunteers. It raised antitoxin titres to both toxin components four-fold in serum samples and seven-fold in jejunal aspirates over preimmunisation control titres. Jejunal aspirates taken after immunisation from vaccinees, but not controls, neutralised the secretory activity of both toxins in appropriate biological assays. These findings show that synthetically produced vaccines are immunogenic in human beings. The peptide used is a promising vaccine for diarrhoeal disease caused by enterotoxigenic strains of E coli.
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A sucrose density gradient assay was used to demonstrate the specificity and saturation of the binding of [(125)I]cholera toxin to isolated intestinal microvillous membranes from rat small intestine. When the toxin is first complexed to antitoxin and then exposed to intestinal membranes, the binding of cholera toxin is inhibited. To emphasize the physiologic importance of these observations, similar concentrations of [(125)I]cholera toxin were shown to stimulate the accumulation of cyclic AMP in mucosal homogenates and to increase the secretion of fluid into intestinal loops, whereas the same concentrations of toxin mixed with antitoxin had no effect on cyclic AMP accumulation. These studies suggest that cholera toxin attaches to brush border binding sites before exerting its biologic effect and that local intestinal antibody protection against cholera toxin may be due to inhibition of toxin attachment to these binding sites.
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