Microfluidic platform for characterizing TCR–pMHC interactions
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BIOMGB-000011-064103_1.pdf
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Author(s)
Stockslager, Max Andrew
Hecht, Vivian Chaya
Hu, Kevin
Aranda-Michel, Edgar C.
Kimmerling, Robert John
Manalis, Scott R
Shaw, Josephine
Payer, Kristofor Robert
Date Issued
November 2017
Journal
Biomicrofluidics
Publisher
AIP Publishing
Citation
Stockslager, Max A., Josephine Shaw Bagnall, Vivian C. Hecht, Kevin Hu, Edgar Aranda-Michel, Kristofor Payer, Robert J. Kimmerling, and Scott R. Manalis. “Microfluidic Platform for Characterizing TCR–pMHC Interactions.” Biomicrofluidics 11, no. 6 (November 2017): 064103.
Version
Final published version
Abstract
The physical characteristics of the T cell receptor (TCR)-peptide-major histocompatibility complex (pMHC) interaction are known to play a central role in determining T cell function in the initial stages of the adaptive immune response. State-of-the-art assays can probe the kinetics of this interaction with single-molecularbond resolution, but this precision typically comes at the cost of low throughput, since the complexity of these measurements largely precludes "scaling up." Here, we explore the feasibility of detecting specific TCR-pMHC interactions by flowing T cells past immobilized pMHC and measuring the reduction in cell speed due to the mechanical force of the receptor-ligand interaction. To test this new fluidic measurement modality, we fabricated a microfluidic device in which pMHC-coated beads are immobilized in hydrodynamic traps along the length of a serpentine channel. As T cells flow past the immobilized beads, their change in speed is tracked via microscopy. We validated this approach using two model systems: primary CD8+ T cells from an OT-1 TCR transgenic mouse with beads conjugated with H-2Kb:SIINFEKL, and Jurkat T cells with beads conjugated with anti-CD3 and anti-CD28 antibodies.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Mechanical Engineering
Massachusetts Institute of Technology. Microsystems Technology Laboratories
Koch Institute for Integrative Cancer Research at MIT
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1063/1.5002116
