Mass Cytometry Phenotyping of Human Granulocytes Reveals Novel Basophil Functional Heterogeneity
Nora Vivanco Gonzalez,
John-Paul Oliveria,
Dmitry Tebaykin,
Geoffrey T. Ivison,
Kaori Mukai,
Mindy M. Tsai,
Luciene Borges,
Kari C. Nadeau,
Stephen J. Galli,
Albert G. Tsai,
Sean C. Bendall
Affiliations
Nora Vivanco Gonzalez
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA
John-Paul Oliveria
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA; Department of Medicine, Division of Respirology, McMaster University, Hamilton, ON, L8S4K1, Canada
Dmitry Tebaykin
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA
Geoffrey T. Ivison
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA
Kaori Mukai
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA; Sean N. Parker Center for Allergy Research, School of Medicine, Stanford University, Palo Alto, CA 94305, USA
Mindy M. Tsai
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA; Sean N. Parker Center for Allergy Research, School of Medicine, Stanford University, Palo Alto, CA 94305, USA
Luciene Borges
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA
Kari C. Nadeau
Sean N. Parker Center for Allergy Research, School of Medicine, Stanford University, Palo Alto, CA 94305, USA; Department of Medicine, Division of Pulmonary and Critical Care Medicine, School of Medicine, Stanford University, Palo Alto, CA 94305, USA
Stephen J. Galli
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA; Sean N. Parker Center for Allergy Research, School of Medicine, Stanford University, Palo Alto, CA 94305, USA; Department of Microbiology and Immunology, School of Medicine, Stanford University, Palo Alto, CA 94305, USA
Albert G. Tsai
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA
Sean C. Bendall
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA; Corresponding author
Summary: Basophils, the rarest granulocyte, play critical roles in parasite- and allergen-induced inflammation. We applied mass cytometry (CyTOF) to simultaneously asses 44 proteins to phenotype and functionally characterize neutrophils, eosinophils, and basophils from 19 healthy donors. There was minimal heterogeneity seen in eosinophils and neutrophils, but data-driven analyses revealed four unique subpopulations within phenotypically basophilic granulocytes (PBG; CD45+HLA-DR−CD123+). Through CyTOF and fluorescence-activated cell sorting (FACS), we classified these four PBG subpopulations as (I) CD16lowFcεRIhighCD244high (88.5 ± 1.2%), (II) CD16highFcεRIhighCD244high (9.1 ± 0.4%), (III) CD16lowFcεRIlowCD244low (2.3 ± 1.3), and (IV) CD16highFcεRIlowCD244low (0.4 ± 0.1%). Prospective isolation confirmed basophilic-morphology of PBG I–III, but neutrophilic-morphology of PBG IV. Functional interrogation via IgE-crosslinking or IL-3 stimulation demonstrated that PBG I–II had significant increases in CD203c expression, whereas PBG III–IV remained unchanged compared with media-alone conditions. Thus, PBG III–IV could serve roles in non-IgE-mediated immunity. Our findings offer new perspectives in human basophil heterogeneity and the varying functional potential of these new subsets in health and disease.
