Quantification of A. fumigatus-specific CD154+ T-cells-preanalytic considerations

Med Mycol. 2017 Feb 1;55(2):223-227. doi: 10.1093/mmy/myw054. Epub 2016 Aug 2.

Abstract

Fungal specific CD154+ T-cells have been described as a biomarker in invasive aspergillosis. The influence of sample storage on the detection of these cells was assessed. Six-hour delay prior to PBMC isolation is associated with an 18% decrease of cell viability and alterations of the cellular composition of the sample. This results in 87% reduction of CD154+ A. fumigatus specific cells due to reduced assay sensitivity and increased background values in unstimulated samples. If prompt cell measurement is not feasible, isolated PBMCs can be frozen (at -20°C and -80°C) and processed later with comparable assay reliability (mean value fresh vs. thawing: 0.126, 0.133; Pearson-Coefficient: 0.962).

Keywords: Aspergillus fumigatus; Biomarker; Immune response; Immunology.

MeSH terms

  • Aspergillus fumigatus / immunology*
  • CD40 Ligand / analysis*
  • Cell Survival
  • Female
  • Freezing
  • Humans
  • Invasive Pulmonary Aspergillosis / diagnosis*
  • Lymphocyte Count
  • Male
  • Preservation, Biological
  • Specimen Handling / methods*
  • T-Lymphocyte Subsets / chemistry*
  • T-Lymphocyte Subsets / immunology*
  • Temperature
  • Time Factors
  • Young Adult

Substances

  • CD40 Ligand