Multiplexed imaging of immune cells in staged multiple sclerosis lesions by mass cytometry

  1. Valeria Ramaglia  Is a corresponding author
  2. Salma Sheikh-Mohamed
  3. Karen Legg
  4. Calvin Park
  5. Olga L Rojas
  6. Stephanie Zandee
  7. Fred Fu
  8. Olga Ornatsky
  9. Eric C Swanson
  10. David Pitt
  11. Alexandre Prat
  12. Trevor D McKee
  13. Jennifer L Gommerman
  1. University of Toronto, Canada
  2. Yale School of Medicine, United States
  3. Université de Montreal, Canada
  4. University Health Network, Canada
  5. Fluidigm Inc, Canada
  6. Université de Montréal, Canada

Abstract

Multiple Sclerosis (MS) is characterized by demyelinated and inflammatory lesions in the brain and spinal cord that are highly variable in terms of cellular content. Here we used imaging mass cytometry (IMC) to enable the simultaneous imaging of 15+ proteins within staged MS lesions. To test the potential for IMC to discriminate between different types of lesions, we selected a case with severe rebound MS disease activity after natalizumab cessation. With post-acquisition analysis pipelines we were able to: (1) Discriminate demyelinating macrophages from the resident microglial pool; (2) Determine which types of lymphocytes reside closest to blood vessels; (3) Identify multiple subsets of T and B cells, and (4) Ascertain dynamics of T cell phenotypes vis-à-vis lesion type and location. We propose that IMC will enable a comprehensive analysis of single-cell phenotypes, their functional states and cell-cell interactions in relation to lesion morphometry and demyelinating activity in MS patients.

Data availability

All data generated and analysed during this study are included in the manuscript and supporting files. Source data file has been provided for Figure 7.

Article and author information

Author details

  1. Valeria Ramaglia

    Department of Immunology, University of Toronto, Toronto, Canada
    For correspondence
    v.ramaglia@utoronto.ca
    Competing interests
    Valeria Ramaglia, received a consulting honorarium from EMD Serono.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9401-5988
  2. Salma Sheikh-Mohamed

    Department of Immunology, University of Toronto, Toronto, Canada
    Competing interests
    No competing interests declared.
  3. Karen Legg

    Department of Immunology, University of Toronto, Toronto, Canada
    Competing interests
    No competing interests declared.
  4. Calvin Park

    Department of Neurology, Yale School of Medicine, New Haven, United States
    Competing interests
    No competing interests declared.
  5. Olga L Rojas

    Department of Immunology, University of Toronto, Toronto, Canada
    Competing interests
    No competing interests declared.
  6. Stephanie Zandee

    Department of Neuroscience, Université de Montreal, Montreal, Canada
    Competing interests
    No competing interests declared.
  7. Fred Fu

    STTARR Innovation center, University Health Network, Toronto, Canada
    Competing interests
    No competing interests declared.
  8. Olga Ornatsky

    Fluidigm Inc, Markham, Canada
    Competing interests
    Olga Ornatsky, is an employee of Fluidigm Inc.
  9. Eric C Swanson

    Fluidigm Inc, Markham, Canada
    Competing interests
    Eric C Swanson, is an employee of Fluidigm Inc.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8454-1207
  10. David Pitt

    Department of Neurology, Yale School of Medicine, New Haven, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6407-9542
  11. Alexandre Prat

    Department of Neurosciences, Faculty of Medicine, Université de Montréal, Montreal, Canada
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6188-0580
  12. Trevor D McKee

    STTARR Innovation center, University Health Network, Toronto, Canada
    Competing interests
    No competing interests declared.
  13. Jennifer L Gommerman

    Department of Immunology, University of Toronto, Toronto, Canada
    Competing interests
    Jennifer L Gommerman, is a consultant for Roche (Canada) and currently holds grants with Novartis, EMD Serono, and Roche.

Funding

National Multiple Sclerosis Society (RR-1602-07777)

  • Valeria Ramaglia

Multiple Sclerosis Society of Canada

  • Jennifer L Gommerman

Multiple Sclerosis Society of Canada

  • Alexandre Prat

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: This work included the use of post-mortem brain tissue. Written consent for post-mortem donation of the CNS to research from the MS donor was obtained (ethics committee approval number BH.07.001).Ethical approval for the use of post-mortem brain tissue from the control donor of the Netherlands Brain Bank was obtained (VU Medical Center ethic committee approval Reference number 2009/148)

Reviewing Editor

  1. Isaac M Chiu, Harvard Medical School, United States

Publication history

  1. Received: April 29, 2019
  2. Accepted: August 1, 2019
  3. Accepted Manuscript published: August 1, 2019 (version 1)
  4. Version of Record published: August 23, 2019 (version 2)

Copyright

© 2019, Ramaglia et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Valeria Ramaglia
  2. Salma Sheikh-Mohamed
  3. Karen Legg
  4. Calvin Park
  5. Olga L Rojas
  6. Stephanie Zandee
  7. Fred Fu
  8. Olga Ornatsky
  9. Eric C Swanson
  10. David Pitt
  11. Alexandre Prat
  12. Trevor D McKee
  13. Jennifer L Gommerman
(2019)
Multiplexed imaging of immune cells in staged multiple sclerosis lesions by mass cytometry
eLife 8:e48051.
https://doi.org/10.7554/eLife.48051

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