Proportions of interferon-γ-producing ascites lymphocytes in response to mycobacterial antigens: A help for early diagnosis of peritoneal tuberculosis in a low TB incidence country

PLoS One. 2019 Apr 4;14(4):e0214333. doi: 10.1371/journal.pone.0214333. eCollection 2019.

Abstract

Background: Peritoneal tuberculosis (TB) remains difficult to diagnose because of its non-specific clinical features and the lack of efficient microbiological tests. As delayed diagnosis is associated with high mortality rates, new diagnostic tools are needed.

Methods and findings: We investigated for 24 patients prospectively enrolled with a possible diagnosis of peritoneal TB, the diagnostic value of the analysis of IFN-γ production by peritoneal fluid lymphocytes in response to a short in vitro stimulation with mycobacterial antigens. The patients were classified in two groups: non-TB and confirmed or highly probable TB. Diagnosis of TB was based on microbiological and histopathological criteria and/or a favorable response to anti-TB treatment. The IFN-γ production by peritoneal CD4+ T lymphocytes was analyzed by flow cytometry after an overnight in vitro stimulation with three different mycobacterial antigens, purified protein derivative (PPD), heparin-binding haemagglutinin (HBHA) or early-secreted-antigen-target-6 (ESAT-6). The percentages of PPD-, HBHA- or ESAT-6-induced IFN-γ-producing peritoneal fluid CD4+ T lymphocytes were higher in the TB group than in the non-TB group (p = 0.0007, p = 0.0004, and p = 0.0002 respectively). Based on cut-off values determined by ROC curve analysis of the results from TB and highly probable TB compared to those of non-TB patients, the sensitivity of these three tests was 100% with a specificity of 92%.

Conclusions: The analysis of mycobacterial-induced IFN-γ production by peritoneal lymphocytes is a promising tool to reliably and rapidly diagnose peritoneal TB. Further studies should be performed on larger cohorts of patients in high-TB-incidence countries to confirm the clinical value of this new diagnostic approach for peritoneal TB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Antigens, Bacterial / immunology*
  • Ascites / metabolism*
  • Bacterial Proteins / immunology
  • Belgium / epidemiology
  • CD4-Positive T-Lymphocytes / immunology*
  • Female
  • Humans
  • Incidence
  • Interferon-gamma / biosynthesis*
  • Lectins / immunology
  • Male
  • Middle Aged
  • Mycobacterium tuberculosis / immunology*
  • Peritoneum / pathology*
  • ROC Curve
  • Tuberculin / immunology
  • Tuberculosis / diagnosis*
  • Tuberculosis / epidemiology*
  • Tuberculosis / immunology
  • Tuberculosis / microbiology

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • ESAT-6 protein, Mycobacterium tuberculosis
  • Lectins
  • Tuberculin
  • heparin-binding hemagglutinin
  • Interferon-gamma

Grants and funding

This work was supported by: F.M., Fonds National de la Recherche Scientifique (FNRS) – PDR T.0147.13; F.M., the European Community within the Horizon2020 program TBVAC2020- grant agreement 643381; and F.M., the “Région de Bruxelles-Capitale – Innoviris". The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.