title: Cytometric detection of antigen-specific IFN-gamma/IL-2 secreting cells in the diagnosis of tuberculosis.
abstract: BACKGROUND
  The purpose of this study was to further characterize the immune response to Mycobacterium tuberculosis (Mtb) antigens, in order to provide new insight into host-pathogen interactions in tuberculosis (TB), and to offer tools for a more accurate diagnosis of the different stages of TB.
  
  
  METHODS
  T-cell responses to Bacillus Calmette-Guérin (BCG), purified protein derivative (PPD), early secretory antigenic target-6 (ESAT-6) protein and culture filtrate protein-10 kDa (CFP-10) were measured in terms of interferon (IFN)-gamma and interleukin (IL)-2 release, using a novel flow cytometric cell-secreting cytokine detection technique. The study was conducted on peripheral blood mononuclear cells (PBMC) obtained from active TB patients, latently TB infected individuals, and healthy donors. IL-10 and IL-17 were also measured to test their possible role as indicators of disease activity.
  
  
  RESULTS
  We confirmed that the enumeration of IFN-gamma releasing cells upon Mtb-specific stimulation is sufficient to identify TB patients and that CD8+ T cells concur to IFN-gamma secretion. IL-2 secreting cells were more frequently observed in latent TB infected individuals compared to active TB patients, suggesting that measurement of cells secreting this cytokine could be a marker of disease stage. No discriminating role was associated to IL-10 and IL-17 release in TB patients.
  
  
  CONCLUSIONS
  Our data indicate that the flow cytometric cytokine-secreting cell detection technique may be envisaged as an additional tool for TB diagnosis allowing the analysis of the immune response to M. tuberculosis-related antigens in the different stages of TB.
authors: Dipartimento di Malattie Infettive, Parassitarie e Immunomediate, Istituto Superiore di Sanità, 00161 Roma, Italy. valeria.sargentini@iss.it
authors: Valeria
authors: V
authors: Sargentini
authors: Sabrina
authors: S
authors: Mariotti
authors: Stefania
authors: S
authors: Carrara
authors: Maria Cristina
authors: MC
authors: Gagliardi
authors: Raffaela
authors: R
authors: Teloni
authors: Delia
authors: D
authors: Goletti
authors: Roberto
authors: R
authors: Nisini
mesh_terms: D000328:Adult; D000942:Antigens, Bacterial / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D054884:Host-Pathogen Interactions; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism*; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D020381:Interleukin-17 / Q000276:immunology / Q000378:metabolism; D007376:Interleukin-2 / Q000276:immunology / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000276:immunology; D014376:Tuberculosis / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D020381:Interleukin-17; D007376:Interleukin-2; D016753:Interleukin-10; C072681:Mycobacterium tuberculosis antigens; D007371:Interferon-gamma