A postgenomic approach to identification of Mycobacterium leprae-specific peptides as T-cell reagents.
To identify Mycobacterium leprae-specific human T-cell epitopes, which could be used to distinguish exposure to M. leprae from exposure to Mycobacterium tuberculosis or to environmental mycobacteria or from immune responses following Mycobacterium bovis BCG vaccination, 15-mer synthetic peptides were synthesized based on data from the M. leprae genome, each peptide containing three or more predicted HLA-DR binding motifs. Eighty-one peptides from 33 genes were tested for their ability to induce T-cell responses, using peripheral blood mononuclear cells (PBMC) from tuberculoid leprosy patients (n = 59) and healthy leprosy contacts (n = 53) from Brazil, Ethiopia, Nepal, and Pakistan and 20 United Kingdom blood bank donors. Gamma interferon (IFN-gamma) secretion proved more sensitive for detection of PBMC responses to peptides than did lymphocyte proliferation. Many of the peptides giving the strongest responses in leprosy donors compared to subjects from the United Kingdom, where leprosy is not endemic, have identical, or almost identical, sequences in M. leprae and M. tuberculosis and would not be suitable as diagnostic tools. Most of the peptides recognized by United Kingdom donors showed promiscuous recognition by subjects expressing differing HLA-DR types. The majority of the novel T-cell epitopes identified came from proteins not previously recognized as immune targets, many of which are cytosolic enzymes. Fifteen of the tested peptides had > or =5 of 15 amino acid mismatches between the equivalent M. leprae and M. tuberculosis sequences; of these, eight gave specificities of > or =90% (percentage of United Kingdom donors who were nonresponders for IFN-gamma secretion), with sensitivities (percentage of responders) ranging from 19 to 47% for tuberculoid leprosy patients and 21 to 64% for healthy leprosy contacts. A pool of such peptides, formulated as a skin test reagent, could be used to monitor exposure to leprosy or as an aid to early diagnosis.
Dockrell
H M
HM
Immunology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom. hazel.dockrell@lshtm.ac.uk
Brahmbhatt
S
S
Robertson
B D
BD
Britton
S
S
Fruth
U
U
Gebre
N
N
Hunegnaw
M
M
Hussain
R
R
Manandhar
R
R
Murillo
L
L
Pessolani
M C
MC
Roche
P
P
Salgado
J L
JL
Sampaio
E
E
Shahid
F
F
Thole
J E
JE
Young
D B
DB
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D016680:Genome, Bacterial; D006684:HLA-DR Antigens / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015441:Leprosy, Tuberculoid / Q000175:diagnosis / Q000276:immunology* / Q000382:microbiology; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D013045:Species Specificity; D013601:T-Lymphocytes / Q000276:immunology
D000942:Antigens, Bacterial; D018984:Epitopes, T-Lymphocyte; D006684:HLA-DR Antigens; D010455:Peptides