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Applied and Environmental Microbiology, November 2003, p. 6620-6627, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6620-6627.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of a Lactococcus lactis Strain That Secretes a Major Epitope of Bovine Beta-Lactoglobulin and Evaluation of Its Immunogenicity in Mice

Jean-Marc Chatel,1 Sebastien Nouaille,2 Karine Adel-Patient,1 Yves Le Loir,2 Herman Boe,1 Alexandra Gruss,2 Jean-Michel Wal,1 and Philippe Langella2*

Unité d'Immuno-Allergie Alimentaire, INRA/CEA, CE de Saclay, DRM-SPI, 91191 Gif-sur-Yvette,1 Unité de Recherches Laitières et de Génétique Appliquée, INRA, 78352 Jouy en JosasFrance2

Received 7 April 2003/ Accepted 27 August 2003

Bovine ß-lactoglobulin (Blg) is one of the major cow's milk allergens. Peptide 41-60 of Blg (Blg41-60) was described as a murine T-cell determinant and a murine, rat, and human immunoglobulin E (IgE) epitope. The aim of this study was the expression of Blg41-60 as a fusion protein in the food-grade bacterium Lactococcus lactis and the characterization of its immunogenicity in mice. We constructed a recombinant strain of L. lactis capable of inducible production and secretion of Blg41-60::Nuc, a fusion protein between Blg41-60 and the mature part of the staphylococcal nuclease (Nuc). The highest production yield of Blg41-60::Nuc (32.5 mg/liter) was reached 4 h after induction. At this time, up to 75% of Blg41-60::Nuc was secreted. When monoclonal antibodies specific for Blg41-60 were used, purified Blg41-60::Nuc and synthetic Blg41-60 exhibited very similar immunoreactivities. Subcutaneous coadministration of purified Blg41-60::Nuc and killed nonrecombinant L. lactis resulted in the induction of specific anti-Blg41-60 IgG2a and IgG1. The IgG1/IgG2a ratio and the lack of specific IgE suggest a Th1-type immune response, i.e., a nonallergic response. Similar administrations of the killed Blg41-60::Nuc-producing L. lactis strain did not elicit a specific immune response, whereas a transitory mucosal IgA-specific immune response was induced in mice after oral administration of the live Blg41-60::Nuc-producing L. lactis strain.

* 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 70. Fax: 33(0)1 34 65 20 65. E-mail: langella{at}jouy.inra.fr. {dagger} Present address: Laboratoire de Microbiologie, INRA ENSAR UMR1055, 35042 Rennes, France.

Applied and Environmental Microbiology, November 2003, p. 6620-6627, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6620-6627.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Cortes-Perez, N. G., Ah-Leung, S., Bermudez-Humaran, L. G., Corthier, G., Wal, J.-M., Langella, P., Adel-Patient, K. (2007). Intranasal Coadministration of Live Lactococci Producing Interleukin-12 and a Major Cow's Milk Allergen Inhibits Allergic Reaction in Mice. CVI 14: 226-233 [Abstract] [Full Text]  
  • Hazebrouck, S., Oozeer, R., Adel-Patient, K., Langella, P., Rabot, S., Wal, J.-M., Corthier, G. (2006). Constitutive Delivery of Bovine {beta}-Lactoglobulin to the Digestive Tracts of Gnotobiotic Mice by Engineered Lactobacillus casei. Appl. Environ. Microbiol. 72: 7460-7467 [Abstract] [Full Text]  
  • Guimaraes, V. D., Innocentin, S., Lefevre, F., Azevedo, V., Wal, J.-M., Langella, P., Chatel, J.-M. (2006). Use of Native Lactococci as Vehicles for Delivery of DNA into Mammalian Epithelial Cells. Appl. Environ. Microbiol. 72: 7091-7097 [Abstract] [Full Text]  
  • Crittenden, R. G., Bennett, L. E. (2005). Cow's Milk Allergy: A Complex Disorder. J. Am. Coll. Nutr. 24: 582S-591S [Abstract] [Full Text]  


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