Related Experiment Video
Updated: Jun 6, 2025
Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses
Published on: March 8, 2024
Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the
Ayan Chatterjee1, Aniket Bandyopadhyay2, Tapas Kumar Maiti3
1Advanced Technology Development Centre, Indian Institute of Technology Kharagpur, Kharagpur, India.
Insights
This study introduces a size-selective pairing microfluidic platform for studying immune cell interactions. It reveals rapamycin dampens inflammatory T-cell responses during combined immunotherapy.
Area of Science:
- Immunology
- Cell Biology
- Microfluidics
Background:
- Single-cell analysis is key to understanding immune response heterogeneity.
- Cell size diversity complicates pairing immune cells for study.
- Previous combinatorial immunotherapies show limited clinical success.
Purpose of the Study:
- To develop a microfluidic platform for size-selective pairing (SSP) of immune cells.
- To investigate real-time, single-cell interactions between macrophages and T-cells under combinatorial stimulation.
- To elucidate the role of rapamycin in modulating immune cell communication.
Main Methods:
- Development of a novel microfluidic platform enabling size-selective pairing of single cells with up to a fivefold size difference.
- Real-time monitoring of macrophage-T-cell interactions using fluorescence microscopy.
- Microfluidic sampling and computational analysis of cellular responses to sequential and spatiotemporal stimulations.
- Comparison of single-cell data with bulk-level measurements.
Main Results:
- Achieved over 40% pairing efficiency for cells with significant size disparities using the SSP platform.
- Observed real-time immune synaptic interactions between macrophages and T-cells.
- Identified a regulatory role for rapamycin in dampening inflammatory outputs from T-cells.
- Demonstrated differences between single-cell and bulk-level measurements of immune responses.
Conclusions:
- The SSP platform effectively overcomes size limitations in single-cell pairing for immune cell studies.
- Combinatorial immunotherapeutic stimulation, including rapamycin, influences macrophage-T-cell interactions at the single-cell level.
- Rapamycin acts to suppress inflammatory responses in T-cells, offering insights into optimizing immunotherapies.
Abstract:
Cellular communication at the single-cell level holds immense potential for uncovering response heterogeneity in immune cell behaviors. However, because of significant size diversity among different immune cell types, controlling the pairing of cells with substantial size differences remains a formidable challenge. We developed a microfluidic platform for size-selective pairing (SSP) to pair single cells with up to a fivefold difference in size, achieving over 40% pairing efficiency. We used SSP to investigate the real-time effects of combinatorial immunotherapeutic stimulation on macrophage T-cell interactions at the single-cell level via fluorescence microscopy and microfluidic sampling. While combinatorial activation involving toll-like receptor (TLR) agonists and rapamycin (an mTOR inhibitor) has improved therapeutic efficacy in mice, its clinical success has been limited. Here, we investigated immune synaptic interactions and outcomes at the single-cell level in real time and compared them with bulk-level measurements. Our findings, after tracking and computationally analyzing the effects of sequential and spatiotemporal stimulations of primary mouse macrophages, suggest a regulatory role of rapamycin in dampening inflammatory outputs in T cells.
