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Infection and Immunity, May 2004, p. 2731-2737, Vol. 72, No. 5
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.5.2731-2737.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Enhanced Mucosal Delivery of Antigen with Cell Wall Mutants of Lactic Acid Bacteria

Corinne Grangette,1* Heide Müller-Alouf,1 Pascal Hols,2 Denise Goudercourt,1 Jean Delcour,2 Mireille Turneer,3 and Annick Mercenier1,{dagger}

Laboratoire de Bactériologie des Ecosytèmes, Institut Pasteur de Lille-Institut de Biologie de Lille, 59019 Lille Cedex, France,1 Unité de Génétique, Université catholique de Louvain, B-1348 Louvain-la-Neuve,2 Laboratoire du Tétanos, Institut Pasteur de Bruxelles, B-1180 Brussels, Belgium3

Received 6 November 2003/ Returned for modification 23 December 2003/ Accepted 5 February 2004

The potential of recombinant lactic acid bacteria (LAB) to deliver heterologous antigens to the immune system and to induce protective immunity has been best demonstrated by using the C subunit of tetanus toxin (TTFC) as a model antigen. Two types of LAB carriers have mainly been used, Lactobacillus plantarum and Lactococcus lactis, which differ substantially in their abilities to resist passage through the stomach and to persist in the mouse gastrointestinal tract. Here we analyzed the effect of a deficiency in alanine racemase, an enzyme that participates in cell wall synthesis, in each of these bacterial carriers. Recombinant wild-type and mutant strains of L. plantarum NCIMB8826 and L. lactis MG1363 producing TTFC intracellularly were constructed and used in mouse immunization experiments. Remarkably, we observed that the two cell wall mutant strains were far more immunogenic than their wild-type counterparts when the intragastric route was used. However, intestinal TTFC-specific immunoglobulin A was induced only after immunization with the recombinant L. plantarum mutant strain. Moreover, the alanine racemase mutant of either LAB strain allowed induction of a much stronger serum TTFC-specific immune response after immunization via the vagina, which is a quite different ecosystem than the gastrointestinal tract. The design and use of these mutants thus resulted in a major improvement in the mucosal delivery of antigens exhibiting vaccine properties.

* Corresponding author Mailing address: Laboratoire de Bactériologie des Ecosystèmes, Institut Pasteur de Lille, 1, rue du Pr Calmette, B.P. 245, F-59019 Lille Cedex, France. Phone: (33) 320-87-11-88. Fax: (33) 320-87-11-92. E-mail: corinne.grangette{at}ibl.fr.

Editor: J. N. Weiser

{dagger} Present address: Nutrition and Health Department, Nestle Research Centre, CH 1000 Lausanne, Switzerland.

Infection and Immunity, May 2004, p. 2731-2737, Vol. 72, No. 5
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.5.2731-2737.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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