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T Cell Dynamic Activation and Functional Analysis in Nanoliter Droplet Microarray.

Saheli Sarkar1, Vinny Motwani1, Pooja Sabhachandani1

  • 1Department of Pharmaceutical Sciences, Northeastern University, 360 Huntington Avenue, Boston, 02115 MA, USA.

Journal of Clinical & Cellular Immunology
|November 28, 2015
PubMed
Summary

This study introduces a microfluidic droplet platform for analyzing immune cell responses. The technology reveals dynamic calcium signaling in T cells upon interaction with dendritic cells, highlighting heterogeneity in immune activation.

Keywords:
CalciumDynamicsHeterogeneityImmune responseLymphocytesMicrofluidicsSingle cell analysisTime-lapse microscopy

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Area of Science:

  • Immunology
  • Microfluidics
  • Cellular Biology

Background:

  • Assessing immune response heterogeneity requires dynamic single-cell analysis and cell-cell interaction studies.
  • Current methods struggle to dynamically investigate non-adherent cell interactions and paracrine signaling without interference.

Purpose of the Study:

  • To develop and validate a microfluidic droplet microarray platform for simultaneous analysis of single T cell responses and heterotypic cell pair interactions.
  • To evaluate the dynamic activation state of primary T cells using this novel platform.

Main Methods:

  • Generation of monodisperse nanoliter droplets containing single human T cells or co-encapsulated T cells and dendritic cells.
  • On-chip stimulation with ionomycin and dynamic calcium signaling monitoring.
  • Co-culture of T cells with activated dendritic cells to observe cell-cell contact-dependent and independent signaling.

Main Results:

  • Ionomycin stimulation induced heterogeneous calcium signaling fluctuations in T cells.
  • T cells exhibited immediate calcium signaling upon contact with dendritic cells, indicating early activation.
  • Delayed, non-contact mediated calcium signaling increases were also observed in T cells.

Conclusions:

  • The nanoliter droplet array microfluidic platform enables dynamic assessment of cellular response heterogeneity.
  • This technology facilitates the detection of early and delayed immune signaling events.
  • It offers a promising approach for live cell phenotyping of immune cells.