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Applied and Environmental Microbiology, December 2006, p. 7460-7467, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01032-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Constitutive Delivery of Bovine ß-Lactoglobulin to the Digestive Tracts of Gnotobiotic Mice by Engineered Lactobacillus casei{triangledown}

S. Hazebrouck,1* R. Oozeer,2 K. Adel-Patient,1 P. Langella,2 S. Rabot,2 J.-M. Wal,1 and G. Corthier2

Unité d'Immuno-Allergie Alimentaire, INRA-CEA, CEA de Saclay, 91191 Gif-sur-Yvette,1 Unité d'Ecologie et Physiologie du Système Digestif, INRA, 78352 Jouy-en-Josas, France2

Received 4 May 2006/ Accepted 17 September 2006

The gut microbiota is critical for maturation of the immune system. Recent evidence suggests that early establishment of lactobacilli in the intestinal microbiota, during neonatal colonization or by probiotic supplementation, could prevent the development of allergic disorders. Postnatal maturation of the gut immune system with allergen-producing lactobacilli colonizing the digestive tract could then affect the development of further allergic sensitization. In this paper, we describe construction of a recombinant Lactobacillus casei strain that can constitutively deliver bovine ß-lactoglobulin (BLG), a major cow's milk allergen, to the guts of gnotobiotic mice. The blg gene was inserted into the L. casei chromosome downstream of an endogenous promoter. BLG production was improved by fusing the propeptide LEISSTCDA (LEISS) to the BLG mature moiety. This led to a 10-fold increase in LEISS-BLG production compared to the production obtained without the propeptide and also led to enhanced secretion corresponding to 5% of the total production. After inoculation into germfree C3H/HeN mice, the genetic stability of the recombinant strain and in vivo BLG production were confirmed for at least 10 weeks. BLG stimulation of spleen cells from mice monoassociated with the BLG-producing lactobacilli induced secretion of the Th1 cytokine gamma interferon and, to a lesser extent, the Th2 cytokine interleukin-5. No BLG-specific immunoglobulin G1 (IgG1), IgG2a, or IgA was detected in sera or in fecal samples. These results suggest that gut colonization with allergen-producing lactobacilli could provide a useful model for studying the modulation of allergic disorders.

* Corresponding author. Mailing address: Laboratoire d'Immuno-Allergie Alimentaire, INRA-CEA, CEA de Saclay, SPI-Bât. 136, 91191 Gif-sur-Yvette cedex, France. Phone: 33 (0)1 69 08 45 97. Fax: 33 (0)1 69 08 59 07. E-mail: Stephane.HAZEBROUCK{at}cea.fr.

{triangledown} Published ahead of print on 22 September 2006.

Applied and Environmental Microbiology, December 2006, p. 7460-7467, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01032-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Liu, J.-K., Hou, X.-L., Wei, C.-H., Yu, L.-Y., He, X.-J., Wang, G.-H., Lee, J.-S., Kim, C.-J. (2009). Induction of Immune Responses in Mice after Oral Immunization with Recombinant Lactobacillus casei Strains Expressing Enterotoxigenic Escherichia coli F41 Fimbrial Protein. Appl. Environ. Microbiol. 75: 4491-4497 [Abstract] [Full Text]  


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