Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level

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 remain...

Full description

Saved in:
Bibliographic Details
Main Authors: Ayan Chatterjee, Aniket Bandyopadhyay, Tapas Kumar Maiti, Tarun Kanti Bhattacharyya
Format: Article
Language:English
Published: Nature Publishing Group 2024-11-01
Series:Microsystems & Nanoengineering
Online Access:https://doi.org/10.1038/s41378-024-00769-3
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850064926116151296
author Ayan Chatterjee
Aniket Bandyopadhyay
Tapas Kumar Maiti
Tarun Kanti Bhattacharyya
author_facet Ayan Chatterjee
Aniket Bandyopadhyay
Tapas Kumar Maiti
Tarun Kanti Bhattacharyya
author_sort Ayan Chatterjee
collection DOAJ
description 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.
format Article
id doaj-art-b9bba8aa50d34574a5d64099dc3032a8
institution DOAJ
issn 2055-7434
language English
publishDate 2024-11-01
publisher Nature Publishing Group
record_format Article
series Microsystems & Nanoengineering
spelling doaj-art-b9bba8aa50d34574a5d64099dc3032a82025-08-20T02:49:09ZengNature Publishing GroupMicrosystems & Nanoengineering2055-74342024-11-0110111710.1038/s41378-024-00769-3Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell levelAyan Chatterjee0Aniket Bandyopadhyay1Tapas Kumar Maiti2Tarun Kanti Bhattacharyya3Advanced Technology Development Centre, Indian Institute of Technology KharagpurInstitute of Cell Dynamics and Imaging, University of MünsterDepartment of Biotechnology, Indian Institute of Technology KharagpurAdvanced Technology Development Centre, Indian Institute of Technology KharagpurAbstract 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.https://doi.org/10.1038/s41378-024-00769-3
spellingShingle Ayan Chatterjee
Aniket Bandyopadhyay
Tapas Kumar Maiti
Tarun Kanti Bhattacharyya
Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
Microsystems & Nanoengineering
title Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
title_full Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
title_fullStr Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
title_full_unstemmed Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
title_short Size-selective microfluidics delineate the effects of combinatorial immunotherapy on T-cell response dynamics at the single-cell level
title_sort size selective microfluidics delineate the effects of combinatorial immunotherapy on t cell response dynamics at the single cell level
url https://doi.org/10.1038/s41378-024-00769-3
work_keys_str_mv AT ayanchatterjee sizeselectivemicrofluidicsdelineatetheeffectsofcombinatorialimmunotherapyontcellresponsedynamicsatthesinglecelllevel
AT aniketbandyopadhyay sizeselectivemicrofluidicsdelineatetheeffectsofcombinatorialimmunotherapyontcellresponsedynamicsatthesinglecelllevel
AT tapaskumarmaiti sizeselectivemicrofluidicsdelineatetheeffectsofcombinatorialimmunotherapyontcellresponsedynamicsatthesinglecelllevel
AT tarunkantibhattacharyya sizeselectivemicrofluidicsdelineatetheeffectsofcombinatorialimmunotherapyontcellresponsedynamicsatthesinglecelllevel