Using mass cytometry to analyse human B cell activation by IL-4-treated mast cells — Australasian Cytometry Society
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Using mass cytometry to analyse human B cell activation by IL-4-treated mast cells (24037)

Felix Marsh-Wakefield 1 , Angela Ferguson 1 , Helen McGuire 2 3 , Thomas Ashhurst 4 , Gary Halliday 5 , Scott Byrne 1
  1. School of Computer Science, The University of Sydney, Sydney, NSW, Australia
  2. Translational Immunology Group, The University of Sydney, Sydney, NSW, Australia
  3. Ramaciotti Facility for Human Systems Biology, The University of Sydney, Sydney, NSW, Australia
  4. Vascular Immunology Unit, School of Medical Sciences, Faculty of Medicine and Healthy, The University of Sydney, Sydney, NSW, Australia
  5. Human Cancer & Viral Immunology Laboratory, The University of Sydney, Sydney, NSW, Australia

The ultraviolet (UV) component of sunlight can protect from multiple sclerosis (MS), an autoimmune disease of the central nervous system. In mice, this UV-protection is due, in part to immune suppression via the activation of regulatory B cells. Mast cells also play an important role during UV-induced immune suppression, and these events may be linked. Our lab has previously found murine IL-4-treated mast cells could activate B cells that were functionally immune suppressive. These regulatory B cells were later found to be phenotypically similar to those found in vivo after UV exposure. In this study we have investigated whether a similar event occurs in human mast cells and B cells. Human mast cells, derived from CD34+ stem cells isolated from peripheral blood, were treated with IL-4 before being co-cultured with donor-specific B cells. As there is no standard marker for identifying human regulatory B cells, we adopted a mass cytometry approach to comprehensively analyse the phenotype of the B cells emerging from this co-culture. Using a combination of 35 antibodies, we could identify memory, naïve and transitional B cell subsets and analyse marker expression within each group. Mast cells appeared to shift naïve B cells towards a transitional phenotype. There were marked changes in a number of immune-related molecules, with a mast cell-induced increase in CD23 being one of the most significant. Although the function of these cells remains to be determined, these mass cytometry studies show that mast cells do influence the activation status of human B cells.

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