Direct demonstration of the productive capability of cytokines at the single cell level in lung sarcoidosis using multicolor cytometry

Respiration. 2002;69(2):155-64. doi: 10.1159/000056319.

Abstract

Background: Sarcoidosis is a chronic systemic disorder of unknown etiology characterized by noncaseating epithelioid cell granulomatous lesions, around which an increasing number of CD4+ T cells infiltrate. These CD4+ T cells may release interferon-gamma (IFN-gamma) and interleukin-2 (IL-2). These cytokines are considered to play an important role in pathogenesis of sarcoidosis.

Methods: We employed a modification of Jung's method using multicolor flow cytometry to assess the capability of single cells obtained from bronchoalveolar lavage (BAL) fluid to produce various cytokines. BAL CD4+ T cell production of IFN-gamma, IL-2 and IL-4 after phorbol ester and ionomycin stimulation were studied.

Results: The percentage of IFN-gamma- and IL-2-producing CD4+ T cells was significantly higher in patients with sarcoidosis compared to healthy volunteers [84.7 +/- 7.5 vs. 51.2 +/- 14.8% (p < 0.005), and 75.3 +/- 8.7 vs. 39.8 +/-11.0% (p < 0.001), respectively]. No significant difference in the percentage of IL-4-producing CD4+ T cells was noted (1.2 +/- 0.6 vs. 3.5 +/- 2.6%; not significant), whereas the absolute number of IL-4-producing CD4+ T cells was significantly higher in patients with sarcoidosis compared to healthy volunteers (563.6 +/- 330.2 vs. 50.9 +/- 66.9/ml; p < 0.005). In the IL-4-producing CD4+ T cells, about 80% of cells concomitantly produced IFN-gamma and more than 60% of cells also produced IL-2.

Conclusion: We demonstrate that Th1-like-producing cells are predominant in the CD4+ as well as in the CD8+ T cell subset of patients with sarcoidosis. We for the first time demonstrated concomitant capabilities of BAL CD4+ T cells to produce Th1 and Th2 cytokines at the single cell level by multicolor flow cytometry.

MeSH terms

  • Adult
  • Bronchoalveolar Lavage Fluid
  • CD4 Antigens / metabolism
  • CD8 Antigens / metabolism
  • Cytokines / metabolism*
  • Female
  • Flow Cytometry / methods
  • Humans
  • Interleukin-4 / metabolism
  • Male
  • Middle Aged
  • Sarcoidosis, Pulmonary / metabolism*
  • T-Lymphocyte Subsets / metabolism*

Substances

  • CD4 Antigens
  • CD8 Antigens
  • Cytokines
  • Interleukin-4