Identification by mass spectrometry of CD8(+)-T-cell Mycobacterium tuberculosis epitopes within the Rv0341 gene product.
Identification of Mycobacterium tuberculosis proteins that can provide immunological protection against tuberculosis is essential for the development of a more effective vaccine. To identify new vaccine targets, we have used immunoaffinity chromatography to isolate class I HLA-A*0201-peptide complexes from M. tuberculosis-infected cells and sequenced the isolated peptides by mass spectrometry. From this material, we have identified three peptides derived from a single M. tuberculosis protein that is encoded by the M. tuberculosis Rv0341 gene. Although no known protein encoded by the Rv0341 gene has been described, it is predicted to give rise to a 479-amino-acid protein with a molecular mass of 43.9 kDa. The three peptides identified are all nested and were found to be antigenic, in that they were capable of inducing peptide-specific, CD8(+) T cells from healthy blood donors in vitro and capable of recognizing and lysing M. tuberculosis-infected dendritic cells. This methodology provides a powerful tool for the identification of M. tuberculosis proteins that can be evaluated as potential vaccine candidates.
Flyer
David C
DC
Argonex, Inc., Charlottesville, Virginia 22903, USA.
Ramakrishna
Venkatesh
V
Miller
Cara
C
Myers
Helen
H
McDaniel
Melanie
M
Root
Karen
K
Flournoy
Caroline
C
Engelhard
Victor H
VH
Canaday
David H
DH
Marto
Jarrod A
JA
Ross
Mark M
MM
Hunt
Donald F
DF
Shabanowitz
Jeffrey
J
White
Forest M
FM
D000942:Antigens, Bacterial / Q000276:immunology*; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D020298:U937 Cells
D000942:Antigens, Bacterial; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D010455:Peptides