title: Single-cell mass cytometry of TCR signaling: amplification of small initial differences results in low ERK activation in NOD mice.
abstract: Signaling from the T-cell receptor (TCR) conditions T-cell differentiation and activation, requiring exquisite sensitivity and discrimination. Using mass cytometry, a high-dimensional technique that can probe multiple signaling nodes at the single-cell level, we interrogate TCR signaling dynamics in control C57BL/6 and autoimmunity-prone nonobese diabetic (NOD) mice, which show ineffective ERK activation after TCR triggering. By quantitating signals at multiple steps along the signaling cascade and parsing the phosphorylation level of each node as a function of its predecessors, we show that a small impairment in initial pCD3ζ activation resonates farther down the signaling cascade and results in larger defects in activation of the ERK1/2-S6 and IκBα modules. This nonlinear property of TCR signaling networks, which magnifies small initial differences during signal propagation, also applies in cells from B6 mice activated at different levels of intensity. Impairment in pCD3ζ and pSLP76 is not a feedback consequence of a primary deficiency in ERK activation because no proximal signaling defect was observed in Erk2 KO T cells. These defects, which were manifest at all stages of T-cell differentiation from early thymic pre-T cells to memory T cells, may condition the imbalanced immunoregulation and tolerance in NOD T cells. More generally, this amplification of small initial differences in signal intensity may explain how T cells discriminate between closely related ligands and adopt strongly delineated cell fates.
authors.affiliation: Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
authors.forename: Michael
authors.initials: M
authors.lastname: Mingueneau
authors.affiliation: Department of Biological Sciences, Columbia Initiative for Systems Biology, Columbia University, New York, NY 10027;
authors.forename: Smita
authors.initials: S
authors.lastname: Krishnaswamy
authors.affiliation: Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305; and.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Spitzer
authors.affiliation: Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305; and.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Bendall
authors.affiliation: Molecular Biology Section, Division of Biological Sciences, Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Erica L
authors.initials: EL
authors.lastname: Stone
authors.affiliation: Molecular Biology Section, Division of Biological Sciences, Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Hedrick
authors.affiliation: Department of Biological Sciences, Columbia Initiative for Systems Biology, Columbia University, New York, NY 10027;
authors.forename: Dana
authors.initials: D
authors.lastname: Pe'er
authors.affiliation: Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115; cbdm@hms.harvard.edu.
authors.forename: Diane
authors.initials: D
authors.lastname: Mathis
authors.affiliation: Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305; and.
authors.forename: Garry P
authors.initials: GP
authors.lastname: Nolan
authors.affiliation: Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115; cbdm@hms.harvard.edu.
authors.forename: Christophe
authors.initials: C
authors.lastname: Benoist
mesh_terms: D000818:Animals; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000276:immunology; D004195:Disease Models, Animal; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology*; D014644:Genetic Variation; D020672:I-kappa B Proteins / Q000378:metabolism; D007108:Immune Tolerance; D007111:Immunity, Cellular; D007153:Immunologic Deficiency Syndromes / Q000276:immunology*; D007156:Immunologic Memory; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D038041:Lymphopoiesis; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D019950:Mitogen-Activated Protein Kinase 1 / Q000172:deficiency; D000072000:NF-KappaB Inhibitor alpha; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011948:Receptors, Antigen, T-Cell / Q000032:analysis / Q000502:physiology*; D017634:Self Tolerance; D059010:Single-Cell Analysis / Q000379:methods*; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology
chemical_list: D020672:I-kappa B Proteins; C000606205:Nfkbia protein, mouse; D011948:Receptors, Antigen, T-Cell; D000072000:NF-KappaB Inhibitor alpha; D048049:Extracellular Signal-Regulated MAP Kinases; C535148:Mapk1 protein, mouse; D019950:Mitogen-Activated Protein Kinase 1
keywords: CyTOF; NOD; diabetes; signaling; single-cell