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Infection and Immunity, April 2004, p. 2170-2176, Vol. 72, No. 4
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.4.2170-2176.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cell Envelope Protein PPE68 Contributes to Mycobacterium tuberculosis RD1 Immunogenicity Independently of a 10-Kilodalton Culture Filtrate Protein and ESAT-6

Caroline Demangel,1* Priscille Brodin,1 Paul J. Cockle,2 Roland Brosch,1 Laleh Majlessi,3 Claude Leclerc,3 and Stewart T. Cole1

Unité de Génétique Moléculaire Bactérienne,1 Unité de Biologie des Régulations Immunitaires, INSERM E352, Institut Pasteur, 75724 Paris Cedex 15, France,3 VLA Weybridge, Department of Bacterial Diseases, TB Research Group, Addlestone, Surrey, United Kingdom2

Received 21 October 2003/ Returned for modification 16 December 2003/ Accepted 9 January 2004

The protective efficacy of Mycobacterium bovis BCG can be markedly augmented by stable integration of Mycobacterium tuberculosis genomic region RD1. BCG complemented with RD1 (BCG::RD1) encodes nine additional proteins. Among them, 10-kDa culture filtrate protein (CFP-10) and ESAT-6 (6-kDa early secreted antigenic target) are low-molecular-weight proteins that induce potent Th1 responses. Using pools of synthetic peptides, we have examined the potential immunogenicity of four other RD1 products (PE35, PPE68, Rv3878, and Rv3879c). PPE68, the protein encoded by rv3873, was the only one to elicit gamma interferon (IFN-{gamma})-producing cells in C57BL/6 mice infected with M. tuberculosis. Anti-PPE68 T cells were predominantly raised against an epitope mapped in the N-terminal end of the protein. Importantly, inactivation of rv3873 in BCG::RD1 did not modify CFP-10 and ESAT-6 secretion. Moreover, the generation of IFN-{gamma} responses to these antigens following immunization with BCG::RD1 was independent of PPE68 expression. Taken together, these results show that PPE68 is an immunogenic product of the RD1 region, which does not interfere with the secretion and immunogenicity of CFP-10 and ESAT-6.

* Corresponding author. Mailing address: Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, 28 Rue du Docteur Roux, 75724 Paris Cedex 15, France. Phone: (33) 1 45 68 84 49. Fax: (33) 1 40 61 35 83. E-mail: demangel{at}pasteur.fr.

Editor: S. H. E. Kaufmann

Infection and Immunity, April 2004, p. 2170-2176, Vol. 72, No. 4
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.4.2170-2176.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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