Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2016 Jun 28;113(26):E3599-608. doi: 10.1073/pnas.1515364113. Epub 2016 Jun 14.

Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture.

Author information

1
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.
2
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02215;
3
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139;
4
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA 02142; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142;
5
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.

Abstract

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.

KEYWORDS:

cell pairing; cell–cell interactions; microfluidics; multiparameter assay; single-cell analysis

PMID:
27303033
PMCID:
PMC4932925
DOI:
10.1073/pnas.1515364113
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center