Microfluidic platform for characterizing TCR–pMHC interactions
Author(s)Stockslager, Max Andrew; Bagnall, Josephine W; Hecht, Vivian Chaya; Hu, Kevin; Aranda-Michel, Edgar C.; Payer, Kristofor; Kimmerling, Robert John; Manalis, Scott R; ... Show more Show less
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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.
DepartmentDavid H. Koch Institute for Integrative Cancer Research at MIT; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Mechanical Engineering; Massachusetts Institute of Technology. Microsystems Technology Laboratories
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.
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