RESUMEN
Intracellular pathogens, such as Mycobacterium tuberculosis, reside in the phagosomes of macrophages where antigenic processing is initiated. Mycobacterial antigen-MHC class II complexes are formed within the phagosome and are then trafficked to the cell surface. Interferon-γ (IFN-γ) and interleukin-10 (IL-10) influence the outcome of M. tuberculosis infection; however, the role of these cytokines with regard to the formation of M. tuberculosis peptide-MHC-II complexes remains unknown. We analysed the kinetics and subcellular localization of M. tuberculosis peptide-MHC-II complexes in M. tuberculosis-infected human monocyte-derived macrophages (MDMs) using autologous M. tuberculosis-specific CD4(+) T cells. The MDMs were pre-treated with either IFN-γ or IL-10 and infected with M. tuberculosis. Cells were mechanically homogenized, separated on Percoll density gradients and manually fractionated. The fractions were incubated with autologous M. tuberculosis -specific CD4(+) T cells. Our results demonstrated that in MDMs pre-treated with IFN-γ, M. tuberculosis peptide-MHC-II complexes were detected early mainly in the phagosomal fractions, whereas in the absence of IFN-γ, the complexes were detected in the endosomal fractions. In MDMs pre-treated with IL-10, the M. tuberculosis peptide-MHC-II complexes were retained in the endosomal fractions, and these complexes were not detected in the plasma membrane fractions. The results of immunofluorescence microscopy demonstrated the presence of Ag85B associated with HLA-DR at the cell surface only in the IFN-γ-treated MDMs, suggesting that IFN-γ may accelerate M. tuberculosis antigen processing and presentation at the cell membrane, whereas IL-10 favours the trafficking of Ag85B to vesicles that do not contain LAMP-1. Therefore, IFN-γ and IL-10 play a role in the formation and trafficking of M. tuberculosis peptide-MHC-II complexes.
Asunto(s)
Antígenos Bacterianos/inmunología , Interferón gamma/inmunología , Interleucina-10/inmunología , Fagosomas/inmunología , Línea Celular , Humanos , Mycobacterium tuberculosis/inmunologíaRESUMEN
The response to recombinant 10-KD heat shock protein (HSP of Mycobacterium leprae (rML10) was evaluated by indirect ELISA in sera from leprosy patients, household contacts, tuberculosis patients and healthy controls a leprosy-endemic area in tne North East of Argentina. Some technical parameters were a analyzed: within-assay and between-assay variability, dose-response curves and dectability indexed (specificity and sensitivity) of ELISA applied to measure anti-10kDa antibodies. High levels of these antibodies have already been reported in positive baciloscopy patients; herein we have also demonstrated that tuberculosis patients sera cross-react with this M. leprae antigen. This test seems to have a low sensificity for leporsy detection; it confirms that antibodies against highly conserved HSP antigens are important in the polycional response against mycobacterial epitopes in leprosy as well as in tuberculosis. (AU)
Asunto(s)
Adulto , Persona de Mediana Edad , Anciano , Humanos , RESEARCH SUPPORT, NON-U.S. GOVT , Lepra/inmunología , Tuberculosis/inmunología , Chaperonina 10/inmunología , Mycobacterium leprae/inmunología , Antígenos Bacterianos/inmunología , Anticuerpos Antibacterianos/inmunología , Sensibilidad y Especificidad , Ensayo de Inmunoadsorción Enzimática , Reacciones Cruzadas , Anciano de 80 o más AñosRESUMEN
The response to recombinant 10-KD heat shock protein (HSP of Mycobacterium leprae (rML10) was evaluated by indirect ELISA in sera from leprosy patients, household contacts, tuberculosis patients and healthy controls a leprosy-endemic area in tne North East of Argentina. Some technical parameters were a analyzed: within-assay and between-assay variability, dose-response curves and dectability indexed (specificity and sensitivity) of ELISA applied to measure anti-10kDa antibodies. High levels of these antibodies have already been reported in positive baciloscopy patients; herein we have also demonstrated that tuberculosis patients sera cross-react with this M. leprae antigen. This test seems to have a low sensificity for leporsy detection; it confirms that antibodies against highly conserved HSP antigens are important in the polycional response against mycobacterial epitopes in leprosy as well as in tuberculosis.