title: Profiling lymphocyte interactions at the single-cell level by microfluidic cell pairing.
abstract: Establishing a successful immune response requires cell-cell interactions, where the nature of antigen presentation dictates functional outcomes. Methods to study these interactions, however, suffer from limited throughput and a lack of control over cell pairing. Here we describe a microfluidic platform that achieves high-throughput deterministic pairing of lymphocytes with a defined contact time, thereby allowing accurate assessment of early activation events for each pair in controlled microenvironments. More importantly, the platform allows the capture of dynamic processes and static parameters from both partners simultaneously, thus enabling pairwise-correlated multiparametric profiling of lymphocyte interactions over hundreds of pairs in a single experiment. Using our platform, we characterized early activation dynamics of CD8 T cells (OT-1 and TRP1 transnuclear (TN)) and investigated the extent of heterogeneity in T-cell activation and the correlation of multiple readouts. The results establish our platform as a promising tool for quantitative investigation of lymphocyte interactions.
authors.affiliation: 1] Research Laboratory of Electronics, 50 Vassar Street, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts 02139, USA [2] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology (MIT), 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Burak
authors.initials: B
authors.lastname: Dura
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Stephanie K
authors.initials: SK
authors.lastname: Dougan
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Marta
authors.initials: M
authors.lastname: Barisa
authors.affiliation: 1] Research Laboratory of Electronics, 50 Vassar Street, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts 02139, USA [2] Health Sciences and Technology, Massachusetts Institute of Technology (MIT), 45 Carleton Street, Cambridge, Massachusetts 02139, USA.
authors.forename: Melanie M
authors.initials: MM
authors.lastname: Hoehl
authors.affiliation: Research Laboratory of Electronics, 50 Vassar Street, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts 02139, USA.
authors.forename: Catherine T
authors.initials: CT
authors.lastname: Lo
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Hidde L
authors.initials: HL
authors.lastname: Ploegh
authors.affiliation: 1] Research Laboratory of Electronics, 50 Vassar Street, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts 02139, USA [2] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology (MIT), 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA [3] Microsystems Technology Laboratory, Massachusetts Institute of Technology (MIT), 60 Vassar Street, Cambridge, Massachusetts 02139, USA.
authors.forename: Joel
authors.initials: J
authors.lastname: Voldman
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology*; D002118:Calcium / Q000378:metabolism; D002450:Cell Communication; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation; D003600:Cytosol / Q000378:metabolism; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008213:Lymphocyte Activation; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D046210:Microfluidic Analytical Techniques; D010088:Oxidoreductases / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D015398:Signal Transduction; D059010:Single-Cell Analysis
chemical_list: D008562:Membrane Glycoproteins; D010455:Peptides; D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C079621:Tyrp1 protein, mouse; D002118:Calcium