Investigation of Mast Cell Toll-like receptor 3 in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis and Systemic Mastocytosis patients using the novel application of autoMACS magnetic separation and Flow cytometry — ASN Events
  1. Schedule
  2. Poster Session One & Burnet Oration Cocktail Function
  3. Investigation of Mast Cell Toll-like receptor 3 in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis and Systemic Mastocytosis patients using the novel application of autoMACS magnetic separation and Flow cytometry

Investigation of Mast Cell Toll-like receptor 3 in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis and Systemic Mastocytosis patients using the novel application of autoMACS magnetic separation and Flow cytometry (#157)

Cassandra Balinas 1 2 , Helene Cabanas 1 2 , Natalie Eaton 1 2 , Professor Donald Staines 1 2 , Professor Sonya Marshall-Gradisnik 1 2
  1. Griffith University, Southport, QUEENSLAND, Australia
  2. National Centre for Neuroimmunology and Emerging Diseases, Gold Coast, Queensland, Australia

Viral infections and hypersensitivities are commonly reported by Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) patients. Mast cells uniquely mediate type 1 hypersensitivities and resolve viral infections via toll-like receptor 3 (TLR3). 11 CFS/ME (40.42 ± 10.31), 9 Systemic Mastocytosis (SM) (47.00 ± 10.37) and 12 healthy controls (HC) (36.36 ± 9.88) were included by the Fukuda and International Consensus Criteria case definitions of CFS/ME and the World Health Organisation case definition of SM. Following autoMACS magnetic separation, MCPs were stimulated with 20ug/ml of Polyinosinic:polycytidylic acid (Poly I:C) for 24hr. MCP purity (CD117 and Lin2), maturity (CD34 and FcεRI), interaction receptor and ligands (CD154 and HLA-DR), and SM-specific (CD2 and CD25) markers were measured using flow cytometry. Consistent with our pilot investigation (Nguyen et al., 2016), four maturity MCPs were characterised in each group pre and post Poly I:C stimulation. A significant decrease in HLA-DR+/CD154- expression was found between CFS/ME and SM patients (pre (p<0.001) and post (p<0.05) Poly I:C stimulation), as well as between SM and HC patients pre (p<0.001) Poly I:C stimulation. Interestingly, an observed increase in HLA-DR-/CD154+ expression was found post Poly I:C stimulation in CFS/ME patients compared with SM and HCs. No significant differences in CD2 and CD25 expression were found between groups. This pilot investigation provides a novel methodology to characterise MCPs in a relatively inexpensive, less invasive and more rapid fashion. The significant decrease in HLA-DR+/CD154- expression suggests that CFS/ME patients may not acquire a comparable MC abundance as SM. Conversely, the observed increase in HLA-DR-/CD154+ expression post Poly I:C stimulation in CFS/ME patients suggests possible associations between MCs and B lymphocytes, which may elucidate the hypersensitivities reported by CFS/ME patients during viral infections. Further investigation is required to determine the immunological contribution of MCs in the pathophysiology of CFS/ME.

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