Design of immunogenic peptides from Mycobacterium tuberculosis genes expressed during macrophage infection.
In vitro diagnosis of MTB-infection uses MTB-proteins coded for by genes of the region of differentiation 1 (RD1) of the MTB genome. This study wants to test if proteins preferentially expressed during MTB-intracellular growth might provide new targets for the diagnosis of MTB-infection. To this end seventy-five multiepitopic HLA-promiscuous MTB-peptides were designed by quantitative implemented peptide-binding motif analysis from 3 MTB-protein genes expressed in activated human macrophages (MA), 4 genes expressed during growth in non-activated human macrophages (MN-A), 12 housekeeping genes (HKG) and 6 genes of the RD1 region (RD1) as control. ELISpot for IFN-was performed to measure the responses of PBMCs deriving from 45 patients affected by active tuberculosis and 34 controls. In active-TB patients, the mean response to RD1-derived peptides was higher than that to either MA (p<0.01), MN-A (p<0.008) or HKG (p<0.01) derived peptides. In TST-positive subjects all selected peptides elicited significant IFN-T-cell responses (p<0.02 compared to TST-negatives), but without differences between the subgroups. Further, T-cell responses to RD1 peptides were lower in the 23 active-TB treated patients than in the untreated ones (p<0.01). The response to MA peptides in treated active-TB was higher than when untreated (p<0.01). These results demonstrate that the use of in vitro models of MTB-intracellular infection to select MTB gene products for further in silico and in vitro assessment of their immunogenicity have the potential to identify novel antigens amenable to the design of new tools for diagnosis and monitoring of tuberculosis.
Seghrouchni
Fouad
F
Department of Internal Medicine, University of Rome Tor Vergata, Via Montpellier 1, 00133 Rome, Italy. fseghrouchni@yahoo.fr
Contini
Silvia
S
Markova
Roumiana
R
Drenska
Roumiana
R
Sadki
Khalid
K
Baassi
Larbi
L
Baassi
Larbii
L
Todorova
Yana
Y
Terzieva
Velislava
V
Bocchino
Marialuisa
M
Cappelli
Giulia
G
Altieri
Alfonso Maria
AM
Alma
Mario Giuseppe
MG
Benjouad
Abdelaziz
A
Mariani
Francesca
F
Petrunov
Bogdan
B
Colizzi
Vittorio
V
El Aouad
Rajae
R
Saltini
Cesare
C
Amicosante
Massimo
M
D000328:Adult; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D055985:Latent Tuberculosis / Q000175:diagnosis / Q000276:immunology; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000276:immunology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015415:Biomarkers; D010455:Peptides; D007371:Interferon-gamma