Cytometric Detection of Antigen-Specific IFN-gamma/IL-2 Secreting Cells in the Diagnosis of Tuberculosis

BMC Infect Dis. 2009 Jun 23;9:99. doi: 10.1186/1471-2334-9-99.

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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, Bacterial / immunology
  • CD8-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / metabolism
  • Cells, Cultured
  • Female
  • Flow Cytometry
  • Host-Pathogen Interactions
  • Humans
  • Interferon-gamma / immunology
  • Interferon-gamma / metabolism*
  • Interleukin-10 / immunology
  • Interleukin-10 / metabolism
  • Interleukin-17 / immunology
  • Interleukin-17 / metabolism
  • Interleukin-2 / immunology
  • Interleukin-2 / metabolism*
  • Leukocytes, Mononuclear / immunology
  • Male
  • Middle Aged
  • Mycobacterium tuberculosis / immunology
  • Tuberculosis / diagnosis*
  • Tuberculosis / immunology

Substances

  • Antigens, Bacterial
  • Interleukin-17
  • Interleukin-2
  • Interleukin-10
  • Mycobacterium tuberculosis antigens
  • Interferon-gamma