Publication Date:
2011-05-10
Description:
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.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3273988/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3273988/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Bendall, Sean C -- Simonds, Erin F -- Qiu, Peng -- Amir, El-ad D -- Krutzik, Peter O -- Finck, Rachel -- Bruggner, Robert V -- Melamed, Rachel -- Trejo, Angelica -- Ornatsky, Olga I -- Balderas, Robert S -- Plevritis, Sylvia K -- Sachs, Karen -- Pe'er, Dana -- Tanner, Scott D -- Nolan, Garry P -- 1R01CA130826/CA/NCI NIH HHS/ -- 272200700038C/PHS HHS/ -- 5U54 CA143907/CA/NCI NIH HHS/ -- HHSN268201000034C/HV/NHLBI NIH HHS/ -- N0I-HV-00242/HV/NHLBI NIH HHS/ -- P01 CA034233/CA/NCI NIH HHS/ -- PN2 EY018228/EY/NEI NIH HHS/ -- R01 CA130826/CA/NCI NIH HHS/ -- R01 CA130826-04/CA/NCI NIH HHS/ -- RB2-01592/PHS HHS/ -- U19 AI057229/AI/NIAID NIH HHS/ -- U54 CA149145/CA/NCI NIH HHS/ -- New York, N.Y. -- Science. 2011 May 6;332(6030):687-96. doi: 10.1126/science.1198704.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21551058" target="_blank"〉PubMed〈/a〉
Keywords:
Algorithms
;
Antibodies
;
Antigens, Surface/analysis
;
B-Lymphocytes/drug effects/immunology/metabolism
;
Bone Marrow Cells/cytology/*drug effects/*immunology/metabolism
;
Cytokines/metabolism
;
Dasatinib
;
Flow Cytometry/*methods
;
Hematopoiesis
;
Humans
;
Immunophenotyping
;
Lanthanoid Series Elements
;
Leukocytes, Mononuclear/drug effects/immunology/metabolism
;
Lymphocyte Activation
;
Lymphocyte Subsets/*drug effects/*immunology/metabolism
;
Mass Spectrometry
;
Phosphorylation
;
Protein Kinase Inhibitors/pharmacology
;
Protein-Tyrosine Kinases/antagonists & inhibitors
;
Pyrimidines/*pharmacology
;
*Signal Transduction/drug effects
;
Single-Cell Analysis/*methods
;
T-Lymphocytes/drug effects/immunology/metabolism
;
Thiazoles/*pharmacology
;
Transition Elements
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
