Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture.
Resolving how the early signaling events initiated by cell-cell interactions are transduced into diverse functional outcomes necessitates correlated measurements at various stages. Typical approaches that rely on bulk cocultures and population-wide correlations, however, only reveal these relationships broadly at the population level, not within each individual cell. Here, we present a microfluidics-based cell-cell interaction assay that enables longitudinal investigation of lymphocyte interactions at the single-cell level through microfluidic cell pairing, on-chip culture, and multiparameter assays, and allows recovery of desired cell pairs by micromanipulation for off-chip culture and analyses. Well-defined initiation of interactions enables probing cellular responses from the very onset, permitting single-cell correlation analyses between early signaling dynamics and later-stage functional outcomes within same cells. We demonstrate the utility of this microfluidic assay with natural killer cells interacting with tumor cells, and our findings suggest a possible role for the strength of early calcium signaling in selective coordination of subsequent cytotoxicity and IFN-gamma production. Collectively, our experiments demonstrate that this new approach is well-suited for resolving the relationships between complex immune responses within each individual cell.
Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA 02139; Electrical Engineering and Computer Science Department, Massachusetts Institute of Technology, Cambridge, MA 02139; Microsystems Technology Laboratory, Massachusetts Institute of Technology, Cambridge, MA 02139; dura@mit.edu voldman@mit.edu.
Burak
B
Dura
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02215;
Mariah M
MM
Servos
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139;
Rachel M
RM
Barry
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA 02142; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142;
Hidde L
HL
Ploegh
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02215; Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA 02142; Division of Immunology, Harvard Medical School, Boston, MA 02115.
Stephanie K
SK
Dougan
Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA 02139; Electrical Engineering and Computer Science Department, Massachusetts Institute of Technology, Cambridge, MA 02139; Microsystems Technology Laboratory, Massachusetts Institute of Technology, Cambridge, MA 02139; dura@mit.edu voldman@mit.edu.
Joel
J
Voldman
D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D002450:Cell Communication; D002460:Cell Line; D018920:Coculture Techniques; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007694:Killer Cells, Natural / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*
D007371:Interferon-gamma; D002118:Calcium
cell pairing; cell–cell interactions; microfluidics; multiparameter assay; single-cell analysis