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Infection and Immunity, November 2003, p. 6116-6123, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6116-6123.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

PPE Protein (Rv3873) from DNA Segment RD1 of Mycobacterium tuberculosis: Strong Recognition of Both Specific T-Cell Epitopes and Epitopes Conserved within the PPE Family

Limei Meng Okkels,1* Inger Brock,1 Frank Follmann,1 Else Marie Agger,1 Sandra M. Arend,2 Tom H. M. Ottenhoff,2 Fredrik Oftung,3 Ida Rosenkrands,1 and Peter Andersen1

Department of Infectious Disease Immunology, Statens Serum Institut, DK-2300 Copenhagen, Denmark,1 Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands,2 Norwegian Institute of Public Health, Nydalen, 0403 Oslo, Norway3

Received 2 April 2003/ Returned for modification 26 May 2003/ Accepted 29 July 2003

Proteins encoded by DNA segment RD1 of Mycobacterium tuberculosis have recently been demonstrated to play important roles in bacterial virulence, vaccine development, and diagnostic reagent design. Previously, we characterized two immunodominant T-cell antigens, the early secreted antigen target (ESAT-6) and the 10-kDa culture filtrate protein (CFP10), which are encoded by the esx-lhp operon in this region. In the present study we characterized a third putative open reading frame in this region, rv3873, which encodes a PPE protein. We found that the rv3873 gene is expressed in M. tuberculosis H37Rv and that the native protein, Rv3873, is predominantly associated with the mycobacterial cell or wall. When tested as a His-tagged recombinant protein, Rv3873 stimulated high levels of gamma interferon secretion in peripheral blood mononuclear cells isolated from tuberculosis (TB) patients, as well as from healthy tuberculin purified protein derivative-positive donors. In contrast to other RD1-encoded antigens, Rv3873 was also found to be recognized by a significant proportion of Mycobacterium bovis BCG-vaccinated donors. Epitope mapping performed with overlapping peptides revealed a broad pattern of T-cell recognition comprising both TB-specific epitopes and epitopes also recognized by BCG-vaccinated donors. The immunodominant epitope (residues 118 to 135) for both TB patients and BCG-vaccinated individuals was found to be highly conserved among a large number of PPE family members.

* Corresponding author. Mailing address: Department of Infectious Disease Immunology, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen, Denmark. Phone: 45-32683480. Fax: 45-32683035. E-mail: lmo{at}ssi.dk.

Editor: S. H. E. Kaufmann

Infection and Immunity, November 2003, p. 6116-6123, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6116-6123.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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