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Infection and Immunity, December 1998, p. 5669-5676, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Systemic and Mucosal Immune Responses after Intranasal Administration of Recombinant Mycobacterium bovis Bacillus Calmette-Guérin Expressing Glutathione S-Transferase from Schistosoma haematobium

Laurent Kremer,1,dagger Loïc Dupré,2 Gilles Riveau,2 André Capron,2 and Camille Locht1,*

Laboratoire de Microbiologie Génétique et Moléculaire, INSERM U447,1 and Laboratoire des Relations Hôtes-Parasite et Stratégies Vaccinales, INSERM U167,2 Institut Pasteur de Lille, F-59019 Lille Cedex, France

Received 13 July 1998/Returned for modification 17 August 1998/Accepted 17 September 1998

A major goal of current vaccine development is the induction of strong immune responses against protective antigens delivered by mucosal routes. One of the most promising approaches in that respect relies on the use of live recombinant vaccine carriers. In this study, Mycobacterium bovis BCG was engineered to produce an intracellular glutathione S-transferase from Schistosoma haematobium (Sh28GST). The gene encoding Sh28GST was placed under the control of the mycobacterial hsp60 promoter on a replicative shuttle plasmid containing a mercury resistance operon as the only selectable marker. The recombinant Sh28GST produced in BCG bound glutathione and expressed enzymatic activity, indicating that its active site was properly folded. Both intraperitoneal and intranasal immunizations of BALB/c mice with the recombinant BCG resulted in strong anti-Sh28GST antibody responses, which were enhanced by a boost. Mice immunized intranasally produced a mixed response with the production of Sh28GST-specific immunoglobulin G1 (IgG1), IgG2a, IgG2b, and IgA in the serum. In addition, high levels of anti-Sh28GST IgA were also found in the bronchoalveolar lavage fluids, demonstrating that intranasal delivery of the recombinant BCG was able to induce long-lasting secretory and systemic immune responses to antigens expressed intracellularly. Surprisingly, intranasal immunization with the BCG producing the Sh28GST induced a much stronger specific humoral response than intranasal immunization with BCG producing the glutathione S-transferase from Schistosoma mansoni, although the two antigens have over 90% identity. This difference was not observed after intraperitoneal administration.

* Corresponding author. Mailing address: Laboratoire de Microbiologie Génétique et Moléculaire, INSERM U447, Institut Pasteur de Lille, 1 rue du Professeur Calmette, F-59019 Lille Cedex, France. Phone: (33) 3.20.87.11.51. Fax: (33) 3.20.87.11.58. E-mail: camille.locht{at}pasteur-lille.fr.

dagger Present address: Department of Microbiology, The Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, United Kingdom.

Infection and Immunity, December 1998, p. 5669-5676, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.