title: High-Content Quantification of Single-Cell Immune Dynamics.
abstract: Cells receive time-varying signals from the environment and generate functional responses by secreting their own signaling molecules. Characterizing dynamic input-output relationships in single cells is crucial for understanding and modeling cellular systems. We developed an automated microfluidic system that delivers precisely defined dynamical inputs to individual living cells and simultaneously measures key immune parameters dynamically. Our system combines nanoliter immunoassays, microfluidic input generation, and time-lapse microscopy, enabling study of previously untestable aspects of immunity by measuring time-dependent cytokine secretion and transcription factor activity from single cells stimulated with dynamic inflammatory inputs. Employing this system to analyze macrophage signal processing under pathogen inputs, we found that the dynamics of TNF secretion are highly heterogeneous and surprisingly uncorrelated with the dynamics of NF-κB, the transcription factor controlling TNF production. Computational modeling of the LPS/TLR4 pathway shows that post-transcriptional regulation by TRIF is a key determinant of noisy and uncorrelated TNF secretion dynamics in single macrophages.
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Junkin
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
authors.forename: Alicia J
authors.initials: AJ
authors.lastname: Kaestli
authors.affiliation: Institute for Quantitative and Computational Biosciences and Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90025, USA.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Jordi
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
authors.forename: Cem
authors.initials: C
authors.lastname: Albayrak
authors.affiliation: Institute for Quantitative and Computational Biosciences and Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90025, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Hoffmann
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland; Institute for Molecular Engineering, University of Chicago, Chicago, IL 60637, USA; Institute for Genomics and Systems Biology, University of Chicago, Chicago, IL 60637, USA. Electronic address: tays@uchicago.edu.
authors.forename: Savaş
authors.initials: S
authors.lastname: Tay
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002469:Cell Separation; D002477:Cells / Q000276:immunology*; D002999:Clone Cells; D016207:Cytokines / Q000032:analysis; D005786:Gene Expression Regulation; D056656:Lab-On-A-Chip Devices; D008070:Lipopolysaccharides; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D016328:NF-kappa B / Q000378:metabolism; D015398:Signal Transduction; D059010:Single-Cell Analysis / Q000379:methods*; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D016207:Cytokines; D008070:Lipopolysaccharides; D016328:NF-kappa B; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha
keywords: NF-κB; cytokine; dynamics; input; microfluidic; single-cell analysis