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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 109, No. 8, 2014, pp. 999-1004
Bioline Code: oc14151
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 109, No. 8, 2014, pp. 999-1004

 en Mycobacterium tuberculosis check for this species in other resources epitope-specific interferon-γ production in healthy Brazilians reactive and non-reactive to tuberculin skin test
da Silva, Bosco Christiano Maciel; Grassi, Maria Fernanda Rios; Coutinho, Raimundo; Mascarenhas, Rita Elizabeth Moreira; Olavarria, Viviana Nilla; Coutinho-Borgo, Adriana; Kalil, Jorge; Cunha-Neto, Edecio & Fonseca, Simone Gonçalves

Abstract

The interferon (IFN)-γ response to peptides can be a useful diagnostic marker of Mycobacterium tuberculosis check for this species in other resources (MTB) latent infection. We identified promiscuous and potentially protective CD4+ T-cell epitopes from the most conserved regions of MTB antigenic proteins by scanning the MTB antigenic proteins GroEL2, phosphate-binding protein 1 precursor and 19 kDa antigen with the TEPITOPE algorithm. Seven peptide sequences predicted to bind to multiple human leukocyte antigen (HLA)-DR molecules were synthesised and tested with IFN-γ enzyme-linked immunospot (ELISPOT) assays using peripheral blood mononuclear cells (PBMCs) from 16 Mantoux tuberculin skin test (TST)-positive and 16 TST-negative healthy donors. Eighty-eight percent of TST-positive donors responded to at least one of the peptides, compared to 25% of TST-negative donors. Each individual peptide induced IFN-γ production by PBMCs from at least 31% of the TST-positive donors. The magnitude of the response against all peptides was 182 ± 230 x 106 IFN-γ spot forming cells (SFC) among TST-positive donors and 36 ± 62 x 106 SFC among TST-negative donors (p = 0.007). The response to GroEL2 (463-477) was only observed in the TST-positive group. This combination of novel MTB CD4 T-cell epitopes should be tested in a larger cohort of individuals with latent tuberculosis (TB) to evaluate its potential to diagnose latent TB and it may be included in ELISPOT-based IFN-γ assays to identify individuals with this condition.

Keywords
Mycobacterium tuberculosis; epitopes; IFN-γ ELISPOT; latent tuberculosis; Mantoux tuberculin skin test

 
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