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Applied and Environmental Microbiology, April 2001, p. 1423-1428, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1423-1428.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Bovine Rotavirus Nonstructural Protein 4 Produced by Lactococcus lactis Is Antigenic and Immunogenic

Vincent Enouf,1 Philippe Langella,2,* Jacqueline Commissaire,2 Jean Cohen,1 and Gérard Corthier3

Unité de Virologie et Immunologie Moléculaires,1 Unité de Recherches Laitières et de Génétique Appliquée,2 and Unité d'Ecologie et de Physiologie du Tube Digestif,3 INRA, 78352 Jouy en Josas, France

Received 3 August 2000/Accepted 6 January 2001

Rotavirus nonstructural protein 4 (NSP4) can induce diarrhea in mice. To get insight into the biological effects of NSP4, production of large quantities of this protein is necessary. We first tried to produce the protein in Escherichia coli, but the nsp4 gene proved to be unstable. The capacity of the generally regarded as safe organism Lactococcus lactis to produce NSP4 either intra- or extracellularly was then investigated by using the nisin-controlled expression system. Production of recombinant NSP4 (rNSP4) was observed in L. lactis for both locations. In spite of a very low secretion efficiency, the highest level of production was obtained with the fusion between a lactococcal signal peptide and rNSP4. Cultures of the rNSP4-secreting strain were injected into rabbits, and a specific immune response was elicited. The anti-rNSP4 antibodies produced in these rabbits recognized NSP4 in MA104 cells infected by rotavirus. We showed that L. lactis is able to produce antigenic and immunogenic rNSP4 and thus is a good organism for producing viral antigens.

* Corresponding author. Mailing address: Unité de Recherches Laitières et de Génétique Appliquée, INRA, 78352 Jouy en Josas, France. Phone: 33(0)1 3465 2083. Fax: 33(0)1 3465 2065. E-mail: langella{at}biotec.jouy.inra.fr.

Applied and Environmental Microbiology, April 2001, p. 1423-1428, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1423-1428.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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