title: Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum.
abstract: Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.
authors.affiliation: Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Bendall
authors.forename: Erin F
authors.initials: EF
authors.lastname: Simonds
authors.forename: Peng
authors.initials: P
authors.lastname: Qiu
authors.forename: El-ad D
authors.initials: el-AD
authors.lastname: Amir
authors.forename: Peter O
authors.initials: PO
authors.lastname: Krutzik
authors.forename: Rachel
authors.initials: R
authors.lastname: Finck
authors.forename: Robert V
authors.initials: RV
authors.lastname: Bruggner
authors.forename: Rachel
authors.initials: R
authors.lastname: Melamed
authors.forename: Angelica
authors.initials: A
authors.lastname: Trejo
authors.forename: Olga I
authors.initials: OI
authors.lastname: Ornatsky
authors.forename: Robert S
authors.initials: RS
authors.lastname: Balderas
authors.forename: Sylvia K
authors.initials: SK
authors.lastname: Plevritis
authors.forename: Karen
authors.initials: K
authors.lastname: Sachs
authors.forename: Dana
authors.initials: D
authors.lastname: Pe'er
authors.forename: Scott D
authors.initials: SD
authors.lastname: Tanner
authors.forename: Garry P
authors.initials: GP
authors.lastname: Nolan
mesh_terms: D000465:Algorithms; D000906:Antibodies; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D000069439:Dasatinib; D005434:Flow Cytometry / Q000379:methods*; D006410:Hematopoiesis; D006801:Humans; D016130:Immunophenotyping; D028581:Lanthanoid Series Elements; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D016131:Lymphocyte Subsets / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D013058:Mass Spectrometry; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011743:Pyrimidines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D059010:Single-Cell Analysis / Q000379:methods*; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology*; D028561:Transition Elements
chemical_list: D000906:Antibodies; D000954:Antigens, Surface; D016207:Cytokines; D028581:Lanthanoid Series Elements; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D013844:Thiazoles; D028561:Transition Elements; D011505:Protein-Tyrosine Kinases; D000069439:Dasatinib