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Infection and Immunity, March 2001, p. 1547-1553, Vol. 69, No. 3
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.3.1547-1553.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Mucosal Immune Responses and Protection against Tetanus Toxin after Intranasal Immunization with Recombinant Lactobacillus plantarum

Corinne Grangette,1,* Heide Müller-Alouf,1 Denise Goudercourt,1 Marie-Claude Geoffroy,1 Mireille Turneer,2 and Annick Mercenier1

Département de Microbiologie des Ecosystèmes, Institut Pasteur de Lille, 59019 Lille Cedex, France,1 and Laboratoire du Tétanos, Institut Pasteur de Bruxelles, B-1180 Brussels, Belgium2

Received 2 October 2000/Returned for modification 2 November 2000/Accepted 6 December 2000

The use of live microorganisms as an antigen delivery system is an effective means to elicit local immune responses and thus represents a promising strategy for mucosal vaccination. In this respect, lactic acid bacteria represent an original and attractive approach, as they are safe organisms that are used as food starters and probiotics. To determine whether an immune response could be elicited by intranasal delivery of recombinant lactobacilli, a Lactobacillus plantarum strain of human origin (NCIMB8826) was selected as the expression host. Cytoplasmic production of the 47-kDa fragment C of tetanus toxin (TTFC) was achieved at different levels depending on the plasmid construct. All recombinant strains proved to be immunogenic by the intranasal route in mice and able to elicit very high systemic immunoglobulin G (IgG1, IgG2b, and IgG2a) responses which correlated to the antigen dose. No significant differences in enzyme-linked immunosorbent assay IgG titers were observed when mice were immunized with live or mitomycin C-treated recombinant lactobacilli. Nevertheless, protection against the lethal effect of tetanus toxin was obtained only with the strains producing the highest dose of antigen and was greater following immunization with live bacteria. Significant TTFC-specific mucosal IgA responses were measured in bronchoalveolar lavage fluids, and antigen-specific T-cell responses were detected in cervical lymph nodes, both responses being higher in mice receiving a double dose of bacteria (at a 24-h interval) at each administration. These results demonstrate that recombinant lactobacilli can induce specific humoral (protective) and mucosal antibodies and cellular immune response against protective antigens upon nasal administration.

* Corresponding author. Mailing address: Département de Microbiologie des Ecosystèmes, Institut Pasteur de Lille, 1, rue du Pr Calmette, B.P. 245, F-59019 Lille Cedex, France. Phone: (33) 320-87-77-74. Fax: (33) 320-87-79-08. E-mail: corinne.grangette{at}pasteur-lille.fr.

Infection and Immunity, March 2001, p. 1547-1553, Vol. 69, No. 3
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.3.1547-1553.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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