Mucosal immunogenicity of the Escherichia coli heat-labile enterotoxin: role of the A subunit.
The Escherichia coli heat-labile enterotoxin (LT) is a potent mucosal immunogen, inducing high secretory as well as systemic antibody responses upon oral or intranasal (i.n.) administration. In addition, LT has the capacity to act as an adjuvant in antibody responses against coadministered other antigens. To investigate the role of the individual subunits of LT in the mucosal immunogenicity and adjuvanticity of LT, the LT holotoxin and the non-toxic B subunit (LTB) were cloned separately and purified from overproducing E. coli cultures. Mice were immunized i.n. with the recombinant LT, LTB and combinations of the two and the induction of LTB-specific serum IgG and IgA as well as mucosal S-IgA was monitored. Intranasal administration of 2 micrograms LTB by itself induced a moderate systemic and a low mucosal antibody response, the latter being restricted to the site of immunization. However, addition of a trace amount (50 ng) of LT holotoxin to LTB strongly stimulated both serum antibody and mucosal S-IgA responses to LTB: the antibody levels induced by 2 micrograms LTB supplemented with 50 ng LT were similar to those seen after immunization with 2.9 micrograms of the LT holotoxin alone (representing an amount of 2 micrograms LTB). Furthermore, immunization with LT-supplemented LTB or with LT holotoxin alone, but not immunization with LTB alone, induced an S-IgA response in distant mucosal tissues including the lung, intestine and the urogenital system. Nicking of the LTA chain with trypsin did not enhance the immunogenicity of LT. These results indicate that, although the LTA chain plays an important role in the mucosal immunogenicity of LT including priming of the common mucosal immune system, extremely low amounts of the LT holotoxin suffice for the induction of high antibody responses to LTB, the trace LT and LTB acting in a synergistic fashion.
de Haan
L
L
Department of Physiological Chemistry Groningen Institute for Drug Studies (GIDS), University of Groningen, The Netherlands.
Holtrop
M
M
Verweij
W R
WR
Agsteribbe
E
E
Wilschut
J
J
D000281:Administration, Intranasal; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000917:Antibody Formation / Q000187:drug effects; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology; D004768:Enterotoxins / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology; D029968:Escherichia coli Proteins; D005260:Female; D018928:Immunity, Mucosal / Q000187:drug effects / Q000276:immunology*; D007071:Immunoglobulin A, Secretory / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000097:blood; D008168:Lung / Q000378:metabolism; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D009297:Nasal Mucosa / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology; D013552:Swine
D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D011994:Recombinant Proteins; C027538:heat-labile enterotoxin, E coli