MUTANTS OF ESCHERICHIA-COLI HEAT-LABILE TOXIN LACKING ADP-RIBOSYLTRANSFERASE ACTIVITY ACT AS NONTOXIC, MUCOSAL ADJUVANTS

A nontoxic mutant (LTK7) of the Escherichia coli heat-labile enterotox in (LT) lacking ADP-ribosylating activity but retaining holotoxin form ation was constructed. By using site-directed mutagenesis, the arginin e at position 7 of the A subunit was replaced with lysine. This molecu le, which was nontoxic in several assays, was able to bind to eukaryot ic cells and acted as a mucosal adjuvant for co-administered proteins; BALB/c mice immunized intranasally with LTK7 and ovalbumin developed high levels of serum and local antibodies to ovalbumin and toxin. In a ddition, mice immunized intranasally with fragment C of tetanus toxin and LTK7 were protected against lethal challenge with tetanus toxin. T hus nontoxic mutants of heat-labile toxin can act as effective intrana sal mucosal adjuvants.