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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Immunol. | doi: 10.3389/fimmu.2019.01596

Mass cytometry discovers two discrete subsets of CD39-Treg which discriminate MGUS from Multiple Myeloma

Felix Marsh-Wakefield1, 2, Annabel Kruzins3,  Helen M. McGuire4, 5, 6, Shihong Yang7, Christian E. Bryant7,  Barbara Fazekas De St Groth4, 5, 6,  Najah Nassif3,  Scott N. Byrne1, John Gibson7, 8, Christina Brown7, 8, Stephen Larsen7, 8, Derek McCulloch7, 8, Richard Boyle9,  Georgina Clark10,  Douglas Joshua7, 8, 11, Phoebe J. Ho7, 8, 11 and  Slavica Vuckovic12, 13*
  • 1Discipline of Infectious Diseases and Immunology, Sydney Medical School, University of Sydney, Australia
  • 2Discipline of Pathology, Faculty of Medicine and Health, University of Sydney, Australia
  • 3School of Life Sciences, University of Technology Sydney, Australia
  • 4Ramaciotti Facility for Human Systems Biology, The University of Sydney, Australia
  • 5Discipline of Physiology, Faculty of Medicine and Health, University of Sydney, Australia
  • 6Charles Perkins Centre, University of Sydney, Australia
  • 7Institute of Haematology, Royal Prince Alfred Hospital, Australia
  • 8Sydney Medical School, University of Sydney, Australia
  • 9Department of Orthopaedics, Royal Prince Alfred Hospital, Australia
  • 10Anzac Research Institute, Australia
  • 11Sydney Medical School, University of Sydney, Australia
  • 12Royal Prince Alfred Hospital, Australia
  • 13Faculty of Medicine, The University of Queensland, Australia

Multiple Myeloma (MM) is preceded by the clinically stable condition monoclonal gammopathy of undetermined significance (MGUS), which is defined by the presence of a monoclonal protein and less than 10% malignant plasma cell infiltration in the bone marrow (BM). Critical immune events that discriminate MGUS from newly diagnosed MM (NDMM) patients remain unknown, but may involve changes in the regulatory T cell (Treg) compartment that favour myeloma growth. To address this possibility, we used mass cytometry to interrogate the distribution of multiple subsets within CD25+CD27low/negTreg in matched BM and peripheral blood (PB) of MGUS and NDMM patients. Both mass cytometry and flow cytometry confirmed a dominance of CD39-Treg within the Treg compartment in BM and PB of NDMM patients compared to an even distribution of CD39-Treg and CD39+Treg in MGUS patients. Unsupervised clustering displayed a phenotypic organisation of Treg into 25 metaclusters that confirmed Treg heterogeneity and identified two subsets which emerged exclusively within CD39-Treg of NDMM patients. One subset was found in both BM and PB that phenotypically resembled activated Treg based on CD45RO, CD49d, and CD62L expression; another subset resembled BM-resident Treg based on its tissue-resident phenotype characterised by CD69 expression, a lack of CD62L and CD49d expression and restricted location within the BM. PD-1 and TIGIT coexpressed on activated Treg, while PD-1 was expressed at higher levels on BM-resident Treg then on activated Treg. Both subsets produce Perforin and Granzyme B. In conclusion, the use of mass cytometry discovered two discrete subsets of CD39-Treg which may be permissive of myeloma growth. Understanding the regulatory properties of these subsets may also advance MGUS and MM diagnosis, prognosis, and therapeutic implications for MM patients.

Keywords: MGUS, mm, Mass-cytometry, Treg, FlowSOM

Received: 15 Mar 2019; Accepted: 26 Jun 2019.

Edited by:

Matteo Bellone, San Raffaele Hospital (IRCCS), Italy

Reviewed by:

Irina Maric, Department of Laboratory Medicine, NIH Clinical Center (CC), United States
Roberto Ria, University of Bari Medical School, Italy
Shaji Kumar, Mayo Clinic, United States  

Copyright: © 2019 Marsh-Wakefield, Kruzins, McGuire, Yang, Bryant, Fazekas De St Groth, Nassif, Byrne, Gibson, Brown, Larsen, McCulloch, Boyle, Clark, Joshua, Ho and Vuckovic. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Slavica Vuckovic, Royal Prince Alfred Hospital, Sydney, Australia,