High-Content Quantification of Single-Cell Immune Dynamics.
- Junkin M Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
- Kaestli AJ Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
- Cheng Z Institute for Quantitative and Computational Biosciences and Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90025, USA.
- Jordi C Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
- Albayrak C Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
- Hoffmann A Institute for Quantitative and Computational Biosciences and Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90025, USA.
- Tay S 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.
- 2016-04-07
Published in:
- Cell reports. - 2016
NF-κB
cytokine
dynamics
input
microfluidic
single-cell analysis
3T3 Cells
Animals
Cell Separation
Cells
Clone Cells
Cytokines
Gene Expression Regulation
Lab-On-A-Chip Devices
Lipopolysaccharides
Macrophages
Mice
Models, Biological
NF-kappa B
Signal Transduction
Single-Cell Analysis
Toll-Like Receptor 4
Tumor Necrosis Factor-alpha
English
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.
- Language
-
- English
- Open access status
- gold
- Identifiers
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- DOI 10.1016/j.celrep.2016.03.033
- PMID 27050527
- Persistent URL
- https://folia.unifr.ch/global/documents/36681
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