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J Med Microbiol 56 (2007), 466-474; DOI: 10.1099/jmm.0.46928-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Evaluation of T-cell responses to peptides with MHC class I-binding motifs derived from PE_PGRS 33 protein of Mycobacterium tuberculosis

M. G. Chaitra, M. S. Shaila and R. Nayak

Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India

Correspondence
R. Nayak
nayak{at}mcbl.iisc.ernet.in

Received 5 September 2006
Accepted 12 December 2006

The PE and PPE proteins of Mycobacterium tuberculosis form a source of antigenic variation among different strains of M. tuberculosis. One of the PE_PGRS proteins, Rv1818c, plays a role in the pathogenesis of mycobacterial infection and specifically influences host-cell responses to tuberculosis infection. Although little is known about these two classes of protein, an immunoinformatics approach has indicated the possibility of their participation in eliciting a major histocompatibility complex (MHC) class I-mediated immune response against tuberculosis, as peptides derived from Rv1818c are predicted to bind to MHC class I molecules with high affinity. In the present work, a DNA vaccine was constructed encoding the full-length Rv1818c protein of M. tuberculosis and its immunogenicity was analysed in BALB/c mice. Immunization with Rv1818c DNA induced a strong CD8+ cytotoxic lymphocyte and Th1-type response, with high levels of gamma interferon (IFN-{gamma}) and low levels of interleukin-4. Two nonameric peptides (Peptide6–14 and Peptide385–393) from Rv1818c were identified by their ability to induce the production of IFN-{gamma} by CD8+ T cells in mice immunized with Rv1818c DNA. An epitope-specific response was demonstrated by the lysis of peptide-pulsed antigen-presenting cells, release of cytotoxic granules and IFN-{gamma} production. These peptides bound with high affinity to MHC H-2Kd and showed low dissociation rates of peptide–MHC complexes. These results could form the basis for testing the identified T-cell epitopes of PE_PGRS proteins in the induction of protective immunity against M. tuberculosis challenge in the mouse model.

Abbreviations: BCG, bacille Calmette–Guérin; ELISPOT, enzyme-linked immunosorbent spot assay; IFN-{gamma}, gamma interferon; IL, interleukin; LDH, lactate dehydrogenase; MHC, major histocompatibility complex; PPD, protein-purified derivative; TB, tuberculosis.




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