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Single-Cell Mass Cytometry Of Differential Immune And Drug Responses Across A Human Hematopoietic Continuum
S. Bendall, Erin F. Simonds, P. Qiu, E. Amir, P. Krutzik, R. Finck, R. Bruggner, R. Melamed, A. Trejo, O. Ornatsky, R. S. Balderas, S. Plevritis, K. Sachs, D. Pe’er, S. Tanner, G. Nolan
Published 2011 · Biology, Medicine
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Simultaneous measurement of more than 30 properties in individual human cells is used to characterize signaling in the immune system. 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.
This paper references
Causal Protein-Signaling Networks Derived from Multiparameter Single-Cell Data
K. Sachs (2005)
I D C Fraser (2010)
ITAM-mediated tonic signalling through pre-BCR and BCR complexes
J. Monroe (2006)
Identification of a hierarchy of multipotent hematopoietic progenitors in human cord blood.
R. Majeti (2007)
Cancer stem cells in solid tumors: an overview.
C. O'Brien (2009)
M W Karaman (2008)
THE CANCER CELL
A. Chadli (1963)
M Roederer (1997)
P O Krutzik (2003)
Cell Cycle-Related Cyclin B1 Quantification
P. S. Frisa (2009)
P O Krutzik (2006)
Materials and methods are available as supporting material on Science Online
J G Albeck (2008)
Tyrosine kinase inhibition in diffuse large B-cell lymphoma: molecular basis for antitumor activity and drug resistance of dasatinib
C. Yang (2008)
M Lindauer (2010)
Practical Flow Cytometry
H. Shapiro (1985)
Recent Results of
D B Rosen (2010)
Drug Metab Dispos
M Dohse (2010)
Tyrosine kinase inhibitor-induced platelet dysfunction in patients with chronic myeloid leukemia.
A. Quintás-Cardama (2009)
Single-cell profiling identifies aberrant STAT5 activation in myeloid malignancies with specific clinical and biologic correlates.
N. Kotecha (2008)
Stem Cell Biology: A View toward the Future
I. Lemischka (2005)
Intracellular phospho‐protein staining techniques for flow cytometry: Monitoring single cell signaling events
P. Krutzik (2003)
Dasatinib, a small-molecule protein tyrosine kinase inhibitor, inhibits T-cell activation and proliferation.
A. Schade (2008)
Semin Radiat Oncol
CA O’Brien (2012)
ACTIVE SUPPRESSION OF IMMUNOGLOBULIN ALLOTYPE SYNTHESIS III. IDENTITICATION OF T CELLS AS RESPONSIBLE FOR SUPPRESSION BY CELLS FROM SPLEEN, THYMUS, LYMPH NODE, AND BONE MARROW
L. Herzenberg (1973)
Ann. N.Y. Acad. Sci
I R Lemischka (2005)
AC Tose (1993)
P K Chattopadhyay (2006)
Dasatinib induces complete remission in a patient with primary cerebral involvement of B-cell chronic lymphocytic leukemia failing chemotherapy.
Georg Russwurm (2010)
Immunophenotypic differentiation patterns of normal hematopoiesis in human bone marrow: Reference patterns for age‐related changes and disease‐induced shifts
E. V. van Lochem (2004)
P. Sperryn (1989)
Single Cell Profiling of Potentiated Phospho-Protein Networks in Cancer Cells
J. Irish (2004)
Dynamics of fine T-cell subsets during HIV disease and after thymic ablation by mediastinal irradiation.
M. Roederer (1997)
J. Immunol. Methods
O Ornatsky (2010)
A quantitative analysis of kinase inhibitor selectivity
Mazen W. Karaman (2008)
Study of cell antigens and intracellular DNA by identification of element-containing labels and metallointercalators using inductively coupled plasma mass spectrometry.
O. Ornatsky (2008)
The protean nature of cells in the B lymphocyte lineage.
R. Hardy (2007)
Asymmetric T Lymphocyte Division in the Initiation of Adaptive Immune Responses
J. Chang (2007)
Lise Wilkinson (2005)
B-cell signaling networks reveal a negative prognostic human lymphoma cell subset that emerges during tumor progression
J. Irish (2010)
P Autissier (2010)
J. Exp. Med
L A Herzenberg (1973)
Annu. Rev. Immunol
T Kurosaki (2010)
D R Bandura (2009)
Mol. Cancer Ther
S Nam (2007)
Functional Characterization of FLT3 Receptor Signaling Deregulation in Acute Myeloid Leukemia by Single Cell Network Profiling (SCNP)
D. Rosen (2010)
B lymphocytes: how they develop and function.
T. Lebien (2008)
疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin.
R. Ulevitch (1995)
Int. J. Hematol
K Takano (2000)
S Doulatov (2010)
Nat. Rev. Immunol
J G Monroe (2006)
A High-throughput Quantitative Multiplex Kinase Assay for Monitoring Information Flow in Signaling Networks
Kevin A. Janes (2003)
Evaluation of a 12‐color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans
P. Autissier (2010)
Cytom. B Clin. Cytom
E G Van Lochem (2004)
Navigating the network: signaling cross-talk in hematopoietic cells
Iain D. C. Fraser (2009)
Quantitative analysis of pathways controlling extrinsic apoptosis in single cells.
J. Albeck (2008)
Phenotypes of stem cells from diverse origin
A. Tárnok (2010)
Biology of hematopoietic stem cells and progenitors: implications for clinical application.
M. Kondo (2003)
Clin. Cancer Res
S M Kornblau (2010)
Annu. Rev. Immunol
R J Ulevitch (1995)
Seventeen-colour flow cytometry: unravelling the immune system
S. Perfetto (2004)
The Response of Human Epithelial Cells to TNF Involves an Inducible Autocrine Cascade
Kevin A. Janes (2006)
Cell stem cell.
L. Zon (2007)
Comparison of ATP-Binding Cassette Transporter Interactions with the Tyrosine Kinase Inhibitors Imatinib, Nilotinib, and Dasatinib
M. Dohse (2010)
Novel mutations in the inhibitory adaptor protein LNK drive JAK-STAT signaling in patients with myeloproliferative neoplasms.
S. Oh (2010)
Mack Fulwyler in his own words
J. Paul Robinson (2005)
M Fulwyler (2005)
Mol. Cell. Proteomics
K A Janes (2003)
Annu. Rev. Immunol. (Paris)
M Kondo (2003)
Quantum dot semiconductor nanocrystals for immunophenotyping by polychromatic flow cytometry
P. K. Chattopadhyay (2006)
Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry.
D. Bandura (2009)
Highly multiparametric analysis by mass cytometry.
O. Ornatsky (2010)
Electronic Separation of Biological Cells by Volume
M. Fulwyler (1965)
Whole animal cell sorting of Drosophila embryos.
M. Krasnow (1991)
IL-7 is critical for homeostatic proliferation and survival of naïve T cells
J. T. Tan (2001)
Nat. Chem. Biol
P O Krutzik (2008)
O I Ornatsky (2008)
Eur. J. Clin. Invest
C Sillaber (2009)
Single-cell phospho-specific flow cytometric analysis demonstrates biochemical and functional heterogeneity in human hematopoietic stem and progenitor compartments.
Kenneth D. Gibbs (2011)
Fantl for critical reading of the manuscript. S.C.B. is supported by the Damon Runyon Cancer Research Foundation Fellowship (DRG-2017-09)
P S Frisa (2009)
Annu. Rev. Biochem
R Y Tsien (1998)
Polymer-based elemental tags for sensitive bioassays.
Xudong Lou (2007)
Revised map of the human progenitor hierarchy shows the origin of macrophages and dendritic cells in early lymphoid development
S. Doulatov (2010)
Homeostasis of naive and memory T cells.
C. Surh (2008)
Dasatinib (BMS-354825) inhibits Stat5 signaling associated with apoptosis in chronic myelogenous leukemia cells
S. Nam (2007)
Rapid and prominent up-regulation of high-affinity receptor for immunoglobulin G (Fc gamma RI) by cross-linking of beta 2 integrins on polymorphonuclear leukocytes.
K. Takano (2000)
Dynamic Single-Cell Network Profiles in Acute Myelogenous Leukemia Are Associated with Patient Response to Standard Induction Therapy
S. Kornblau (2010)
Hochhaus , Recent Results
Immunosuppression and atypical infections in CML patients treated with dasatinib at 140 mg daily
C. Sillaber (2009)
Translation of the Philadelphia chromosome into therapy for CML.
B. Druker (2008)
A Tárnok (2010)
Angew. Chem. Int. Ed. Engl
X Lou (2007)
C. Geary (1984)
High-content single-cell drug screening with phosphospecific flow cytometry.
P. Krutzik (2008)
Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling
P. Krutzik (2006)
B cell signaling and fate decision.
T. Kurosaki (2010)
The green fluorescent protein.
R. Tsien (1998)
Nat. Rev. Immunol
S P Perfetto (2004)
This paper is referenced by
The contrasting roles of Th17 immunity in human health and disease
Edwin Hoe (2017)
Guang Li (2015)
Life on a microarray: assessing live cell functions in a microarray format
K. Papp (2012)
Interdisciplinary characterization of T cell dynamics in HIV infection
Marcus Buggert (2014)
Conditional density-based analysis of T cell signaling in single-cell data
Smita Krishnaswamy (2014)
Nikolay Samusik (2017)
Single-cell technologies for monitoring immune systems
P. Chattopadhyay (2014)
Analyzing the phenotypic and functional complexity of lymphocytes using CyTOF (cytometry by time-of-flight)
G. Chen (2012)
Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue Specimens for Enhanced Clinical Guidance
Alexander Borowsky (2015)
HDCytoData: Collection of high-dimensional cytometry benchmark datasets in Bioconductor object formats.
L. M. Weber (2019)
Analysis of the human kinome and phosphatome reveals diseased signaling networks induced by overexpression
Xiao-Kang Lun (2018)
Caractérisation des réponses immunitaires chez les patients atteints de myopathies auto-immunes idiopathiques
Gaëlle Dzangué Tchoupou (2018)
Extracting a Cellular Hierarchy from High-dimensional Cytometry Data with SPADE
P. Qiu (2011)
Single-Cell Immune Competency Signatures Associate with Survival in Phase II GVAX and CRS-207 Randomized Studies in Patients with Metastatic Pancreatic Cancer
N. Nair (2020)
Human Bone Marrow Assessment by Single Cell RNA Sequencing, Mass Cytometry and Flow Cytometry
Karolyn A. Oetjen (2018)
A hardware accelerated system for high throughput cellular image analysis
Dajung Lee (2018)
Multidimensional profiling in the investigation of small-molecule-induced cell death.
Adam J Wolpaw (2014)
Of snowflakes and natural killer cell subsets
L. Lanier (2014)
Systems immunology of human malaria.
T. Tran (2012)
Absolute and Relative Quantification of Multiplex DNA Assays Based on an Elemental Labeling Strategy†
Guojun Han (2013)
Advances in ultrasensitive mass spectrometry of organic molecules.
M. Kandiah (2013)
Nac1 Coordinates a Sub-network of Pluripotency Factors to Regulate Embryonic Stem Cell Differentiation.
Mohan K. Malleshaiah (2016)
Wishbone identifies bifurcating developmental trajectories from single-cell data
M. Setty (2016)
Comparison of Clustering Methods for High-Dimensional Single-Cell Flow and Mass Cytometry Data
L. M. Weber (2016)
Leveraging blood and tissue CD4+ T cell heterogeneity at the single cell level to identify mechanisms of disease in rheumatoid arthritis.
Chamith Y Fonseka (2017)
[Inflammation and osteoclasts].
K. Yokota (2017)
Cell Culture Technology
R. Pörtner (2016)
Systems Immunology: Learning the Rules of the Immune System.
A. Villani (2018)
Single cell proteomics in biomedicine: High‐dimensional data acquisition, visualization, and analysis
Yapeng Su (2017)
Phospho-tuning immunity through Denisovan, modern human and mouse TNFAIP3 gene variants
N. Zammit (2019)
Perturbed myoepithelial cell differentiation in BRCA mutation carriers and in ductal carcinoma in situ
L. Ding (2019)
Deciphering tumor heterogeneity from FFPE tissues: Its promise and challenges
A. Simmons (2017)See more