TY - JOUR T1 - Induction of in vivo functional Db-restricted cytolytic T cell activity against a putative phosphate transport receptor of Mycobacterium tuberculosis. JF - J Immunol Y1 - 2004 A1 - Marta Romano A1 - Olivier J Denis A1 - D'Souza, Sushila A1 - Wang, Xiao-Ming A1 - Ottenhoff, Tom H M A1 - Brulet, Jean-Marc A1 - Huygen, Kris KW - Amino Acid Sequence KW - Animals KW - Antibodies, Bacterial KW - ATP-Binding Cassette Transporters KW - Bacterial Proteins KW - Epitopes, T-Lymphocyte KW - H-2 Antigens KW - Histocompatibility Antigen H-2D KW - Interferon-gamma KW - Interleukin-2 KW - mice KW - Mice, Inbred C57BL KW - Molecular Sequence Data KW - Mycobacterium tuberculosis KW - T-Lymphocytes, Cytotoxic KW - Th1 Cells KW - Tuberculosis Vaccines KW - Vaccines, DNA AB -

Using plasmid vaccination with DNA encoding the putative phosphate transport receptor PstS-3 from Mycobacterium tuberculosis and 36 overlapping 20-mer peptides spanning the entire PstS-3 sequence, we determined the immunodominant Th1-type CD4(+) T cell epitopes in C57BL/10 mice, as measured by spleen cell IL-2 and IFN-gamma production. Furthermore, a potent IFN-gamma-inducing, D(b)-restricted CD8(+) epitope was identified using MHC class I mutant B6.C-H-2(bm13) mice and intracellular IFN-gamma and whole blood CD8(+) T cell tetramer staining. Using adoptive transfer of CFSE-labeled, peptide-pulsed syngeneic spleen cells from naive animals into DNA vaccinated or M. tuberculosis-infected recipients, we demonstrated a functional in vivo CTL activity against this D(b)-restricted PstS-3 epitope. IFN-gamma ELISPOT responses to this epitope were also detected in tuberculosis-infected mice. The CD4(+) and CD8(+) T cell epitopes defined for PstS-3 were completely specific and not recognized in mice vaccinated with either PstS-1 or PstS-2 DNA. The H-2 haplotype exerted a strong influence on immune reactivity to the PstS-3 Ag, and mice of the H-2(b, p, and f) haplotype produced significant Ab and Th1-type cytokine levels, whereas mice of H-2(d, k, r, s, and q) haplotype were completely unreactive. Low responsiveness against PstS-3 in MHC class II mutant B6.C-H-2(bm12) mice could be overcome by DNA vaccination. IFN-gamma-producing CD8(+) T cells could also be detected against the D(b)-restricted epitope in H-2(p) haplotype mice. These results highlight the potential of DNA vaccination for the induction and characterization of CD4(+) and particularly CD8(+) T cell responses against mycobacterial Ags.

VL - 172 CP - 11 U1 - http://www.ncbi.nlm.nih.gov/pubmed/15153510?dopt=Abstract ER -