An automated real-time microfluidic platform to probe single NK cell heterogeneity and cytotoxicity on-chip.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
24 08 2021
24 08 2021
Historique:
received:
05
04
2021
accepted:
03
08
2021
entrez:
25
8
2021
pubmed:
26
8
2021
medline:
3
11
2021
Statut:
epublish
Résumé
Cytotoxicity is a vital effector mechanism used by immune cells to combat pathogens and cancer cells. While conventional cytotoxicity assays rely on averaged end-point measures, crucial insights on the dynamics and heterogeneity of effector and target cell interactions cannot be extracted, emphasizing the need for dynamic single-cell analysis. Here, we present a fully automated droplet-based microfluidic platform that allowed the real-time monitoring of effector-target cell interactions and killing, allowing the screening of over 60,000 droplets identifying 2000 individual cellular interactions monitored over 10 h. During the course of incubation, we observed that the dynamics of cytotoxicity within the Natural Killer (NK) cell population varies significantly over the time. Around 20% of the total NK cells in droplets showed positive cytotoxicity against paired K562 cells, most of which was exhibited within first 4 h of cellular interaction. Using our single cell analysis platform, we demonstrated that the population of NK cells is composed of individual cells with different strength in their effector functions, a behavior masked in conventional studies. Moreover, the versatility of our platform will allow the dynamic and resolved study of interactions between immune cell types and the finding and characterization of functional sub-populations, opening novel ways towards both fundamental and translational research.
Identifiants
pubmed: 34429486
doi: 10.1038/s41598-021-96609-9
pii: 10.1038/s41598-021-96609-9
pmc: PMC8385055
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
17084Informations de copyright
© 2021. The Author(s).
Références
Trends Immunol. 2014 May;35(5):219-29
pubmed: 24746883
Front Immunol. 2012 Oct 05;3:300
pubmed: 23060879
PLoS One. 2020 Apr 1;15(4):e0229520
pubmed: 32236105
PLoS One. 2014 Feb 19;9(2):e89357
pubmed: 24586714
Lab Chip. 2013 Dec 21;13(24):4740-4
pubmed: 24185478
Front Immunol. 2018 Oct 16;9:2373
pubmed: 30459757
J Immunol Methods. 1994 Jun 24;172(2):227-39
pubmed: 7518485
Nat Immunol. 2014 Feb;15(2):128-35
pubmed: 24448570
Nat Immunol. 2008 May;9(5):503-10
pubmed: 18425107
Proc Natl Acad Sci U S A. 2016 Jun 28;113(26):E3599-608
pubmed: 27303033
Nat Protoc. 2020 Sep;15(9):2920-2955
pubmed: 32788719
J Vis Exp. 2019 Feb 11;(144):
pubmed: 30799837
Blood. 2013 Feb 21;121(8):1326-34
pubmed: 23287857
Nat Biotechnol. 2006 Jun;24(6):703-7
pubmed: 16699501
Chem Biol. 2008 May;15(5):427-37
pubmed: 18482695
Trends Biotechnol. 2010 Jun;28(6):281-90
pubmed: 20434785
Nat Biotechnol. 2017 Oct;35(10):977-982
pubmed: 28892076
Clin Diagn Lab Immunol. 2001 Nov;8(6):1131-5
pubmed: 11687452
Immunology. 1968 Feb;14(2):181-96
pubmed: 4966657
Sci Rep. 2018 Jan 24;8(1):1531
pubmed: 29367754
Immunity. 2007 Sep;27(3):393-405
pubmed: 17892848
Integr Biol (Camb). 2014 Dec;6(12):1153-61
pubmed: 25228316
Sci Rep. 2019 Jul 23;9(1):10672
pubmed: 31337806
Nat Commun. 2018 Aug 20;9(1):3317
pubmed: 30127440
J Immunol. 2018 May 1;200(9):3231-3243
pubmed: 29592963
Blood. 2008 Aug 1;112(3):461-9
pubmed: 18650461
Lab Chip. 2008 Jun;8(6):898-906
pubmed: 18497909
Cell Rep. 2020 Apr 28;31(4):107574
pubmed: 32348757
Front Immunol. 2020 Aug 14;11:1851
pubmed: 32922399
Front Immunol. 2016 Apr 05;7:119
pubmed: 27092139
Commun Biol. 2019 Jan 24;2:35
pubmed: 30701200
Lab Chip. 2007 Sep;7(9):1139-45
pubmed: 17713612
PLoS One. 2007 Mar 28;2(3):e326
pubmed: 17389917
Integr Biol (Camb). 2012 Oct;4(10):1175-84
pubmed: 22945136
Front Immunol. 2017 Dec 14;8:1736
pubmed: 29312292
J Immunol Methods. 1983 Nov 25;64(3):313-20
pubmed: 6199426
ACS Omega. 2020 Sep 17;5(38):24674-24683
pubmed: 33015484
Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20173-8
pubmed: 19075237
Proc Natl Acad Sci U S A. 2013 Apr 16;110(16):6488-93
pubmed: 23576740
Lab Chip. 2015 Sep 7;15(17):3439-59
pubmed: 26226550
J Exp Med. 2019 Sep 2;216(9):2113-2127
pubmed: 31270246