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An imaging mass cytometry immunophenotyping panel for non-human primate tissues.

Paula Niewold1, Marieke E Ijsselsteijn2, Frank A W Verreck3

  • 1Department of Infectious Diseases, Leiden University Medical Centre, Leiden, Netherlands.

Frontiers in Immunology
|August 1, 2022
PubMed
Summary

Developing an imaging mass cytometry panel is crucial for understanding cellular organization in non-human primate tissues. This 18-marker panel visualizes leukocyte subsets, aiding research in health and disease outcomes.

Keywords:
cynomolgus macaquedisease model animalimaging mass cytometry (IMC)immunophenotyping (IP)non-human primate (NHP)rhesus macaque (Macaca mulatta)spatial organization

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Area of Science:

  • Cellular Biology
  • Immunology
  • Translational Research

Background:

  • Cellular spatial organization is vital for function in health and disease.
  • Imaging mass cytometry (IMC) preserves spatial context for analyzing cell relationships.
  • Preclinical models, especially non-human primates, are essential for translational research due to physiological similarities to humans.

Purpose of the Study:

  • To develop a pipeline for selecting and incorporating antibodies for imaging mass cytometry on non-human primate formalin-fixed paraffin-embedded (FFPE) tissues.
  • To create a high-dimensional IMC panel for analyzing leukocyte subsets in non-human primate tissues.
  • To advance the understanding of cellular organization and its impact on disease outcomes using non-human primate models.

Main Methods:

  • Development of a novel antibody selection and incorporation pipeline for IMC.
  • Creation of an 18-marker backbone IMC panel.
  • Application of the panel to visualize leukocyte subsets in rhesus and cynomolgus macaque tissues.

Main Results:

  • Successful development of a method for antibody selection for non-human primate FFPE tissue IMC.
  • Establishment of an 18-marker IMC panel.
  • Demonstrated visualization of diverse leukocyte subsets within non-human primate tissues.

Conclusions:

  • The developed pipeline enables effective IMC panel creation for non-human primate tissues.
  • The 18-marker panel facilitates detailed analysis of cellular organization in macaques.
  • This tool will enhance research into disease mechanisms and outcomes in translational studies.