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Infection and Immunity, March 1999, p. 1287-1291, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Zonula Occludens Toxin Is a Powerful Mucosal Adjuvant for Intranasally Delivered Antigens

Mariarosaria Marinaro,1 Annalisa di Tommaso,2 Sergio Uzzau,3 Alessio Fasano,3 and Maria Teresa de Magistris2,*

Laboratory of Bacteriology and Medical Mycology, Istituto Superiore di Sanità, Rome,1 and IRIS, Chiron S.p.A., Siena,2 Italy, and Division of Pediatric Gastroenterology and Nutrition, Center for Vaccine Development, University of Maryland, Baltimore, Maryland3

Received 1 December 1997/Returned for modification 13 January 1998/Accepted 15 December 1998

Zonula occludens toxin (Zot) is produced by toxigenic strains of Vibrio cholerae and has the ability to reversibly alter intestinal epithelial tight junctions, allowing the passage of macromolecules through the mucosal barrier. In the present study, we investigated whether Zot could be exploited to deliver soluble antigens through the nasal mucosa for the induction of antigen-specific systemic and mucosal immune responses. Intranasal immunization of mice with ovalbumin (Ova) and recombinant Zot, either fused to the maltose-binding protein (MBP-Zot) or with a hexahistidine tag (His-Zot), induced anti-Ova serum immunoglobulin G (IgG) titers that were approximately 40-fold higher than those induced by immunization with antigen alone. Interestingly, Zot also stimulated high anti-Ova IgA titers in serum, as well as in vaginal and intestinal secretions. A comparison with Escherichia coli heat-labile enterotoxin (LT) revealed that the adjuvant activity of Zot was only sevenfold lower than that of LT. Moreover, Zot and LT induced similar patterns of Ova-specific IgG subclasses. The subtypes IgG1, IgG2a, and IgG2b were all stimulated, with a predominance of IgG1 and IgG2b. In conclusion, our results highlight Zot as a novel potent mucosal adjuvant of microbial origin.

* Corresponding author. Present address: Istituto Superiore di Sanità, Laboratory of Bacteriology and Medical Mycology, Viale Regina Elena, 299, 00161 Rome, Italy. Phone: (39) 06-4990 2734. Fax: (39) 06-4990 2934. E-mail: mtdemagi{at}iss.it.

Infection and Immunity, March 1999, p. 1287-1291, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.