Novel characterisation of mast cell phenotypes from peripheral blood mononuclear cells in chronic fatigue syndrome/myalgic encephalomyelitis patients

Asian Pac J Allergy Immunol. 2017 Jun;35(2):75-81. doi: 10.12932/ap0771.

Abstract

Background: Mast cells (MCs) mediate inflammation through neuropeptides and cytokines, along with histamine and reactive oxygen species (ROS). Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is an illness characterized by an unexplained disabling fatigue with multiple physiological impairments as well as dysregulated cytokine profiles.

Objective: To determine mast cell phenotypes in isolated human PBMCs, in healthy controls and in CFS/ME patients. Second, determine receptor expression of RAGE and its ligand high mobility group box 1 protein (HMGB1).

Method: Moderately severe CFS/ME patients (n=12, mean age 39.25 ± SD3.52 years), severe CFS/ME patients (n=6, mean age 43.00 ± SD4.02 years) and healthy controls (n=13, mean age 42.69 ± SD3.87 years) were included in this study. CFS/ME patients were classified according to the 2011 International Consensus Criteria. LSRFortessa X-20 Flow cytometry was used for the identification of phenotypic peripheral mast cell population in PBMCs using an exclusion marker Lin2 cocktail (anti-CD3, anti-CD14, anti-CD19, anti-CD20 and anti-CD56) and inclusion markers (CD117, CD34, FCεRI, chymase, HLA-DR and CD154) following comparative investigation. HMGB1 and soluble RAGE expression in plasma was measured by sandwich ELISA assay.

Results: There was a significant increase in CD117⁺CD34⁺FCεRI-chymase- mast cell populations in moderate and severe CFS/ME patients compared with healthy controls. There was a significant increase in CD40 ligand and MHC-II receptors on differentiated mast cell populations in the severe CFS/ME compared with healthy controls and moderate CFS/ME. There were no significant differences between groups for HMGB1 and sRAGE.

Conclusions: This preliminary study investigates mast cell phenotypes from PBMCs in healthy controls. We report significant increase of naïve MCs in moderate and severe CFS/ME patients compared with healthy controls. Moreover, a significant increase in CD40 ligand and MHC-II receptors on differentiated mast cells in severe CFS/ME patients. Peripheral MCs may be present in CFS/ME pathology however, further investigation to determine their role is required.

MeSH terms

  • Adult
  • Antigens, CD / metabolism
  • Biomarkers
  • Case-Control Studies
  • Enzyme-Linked Immunosorbent Assay
  • Fatigue Syndrome, Chronic / immunology*
  • Female
  • Flow Cytometry
  • HMGB1 Protein / blood*
  • Healthy Volunteers
  • Humans
  • Immunophenotyping
  • Leukocytes, Mononuclear / immunology*
  • Male
  • Mast Cells / immunology*
  • Middle Aged
  • Receptor for Advanced Glycation End Products / blood*

Substances

  • AGER protein, human
  • Antigens, CD
  • Biomarkers
  • HMGB1 Protein
  • HMGB1 protein, human
  • Receptor for Advanced Glycation End Products