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Clinical and Diagnostic Laboratory Immunology, May 2001, p. 545-551, Vol. 8, No. 3
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.3.545-551.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Induction of Mucosal Immune Response after Intranasal or Oral Inoculation of Mice with Lactococcus lactis Producing Bovine Beta-Lactoglobulin

Jean-Marc Chatel,1 Philippe Langella,2,* Karine Adel-Patient,1 Jacqueline Commissaire,2 Jean-Michel Wal,1 and Gérard Corthier3

Unité d'Immuno-Allergie Alimentaire INRA-CEA, Service de Pharmacologie et d'Immunologie, Bat 136 CEA Saclay, 91191 Gif sur Yvette,1 and Unité de Recherches Laitières et de Génétique Appliquée2 and Unité d'Ecologie et de Physiologie du Système Digestif,3 INRA, 78352 Jouy en Josas, France

Received 30 May 2000/Returned for modification 19 September 2000/Accepted 6 February 2001

The bovine beta-lactoglobulin (BLG) is a major cow's milk allergen. Here, we evaluated the immune response against BLG induced in mice, using the organism Lactococcus lactis, which has GRAS ("generally regarded as safe") status, as a delivery vehicle. The cDNA of the blg gene, encoding BLG, was expressed and engineered for either intra- or extracellular expression in L. lactis. Using a constitutive promoter, the yield of intracellular recombinant BLG (rBLG) was about 20 ng per ml of culture. To increase the quantity of rBLG, the nisin-inducible expression system was used to produce rBLG in the cytoplasmic and extracellular locations. Although the majority of rBLG remained in the cytoplasm, the highest yield (2 µg per ml of culture) was obtained with a secreting strain that encodes a fusion between a lactococcal signal peptide and rBLG. Whatever the expression system, the rBLG is produced mostly in a soluble, intracellular, and denatured form. The BLG-producing strains were then administered either orally or intranasally to mice, and the immune response to BLG was examined. Specific anti-BLG immunoglobulin A (IgA) antibodies were detected 3 weeks after the immunization protocol in the feces of mice immunized with the secreting lactococcal strain. Specific anti-BLG IgA detected in mice immunized with lactococci was higher than that obtained in mice immunized with the same quantity of pure BLG. No specific anti-BLG IgE, IgA, IgG1, or IgG2a was detected in sera of mice. These recombinant lactococcal strains constitute good vehicles to induce a mucosal immune response to a model allergen and to better understand the mechanism of allergy induced by BLG.

* Corresponding author. Mailing address: Unité de Recherches Laitières et de Génétique Appliquée, INRA, 78352 Jouy en Josas, France. Phone: 33(0)1 34 65 20 83. Fax: 33(0)1 34 65 20 65. E-mail: langella{at}biotec.jouy.inra.fr.

Clinical and Diagnostic Laboratory Immunology, May 2001, p. 545-551, Vol. 8, No. 3
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.3.545-551.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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