Low-Density Granulocyte Contamination From Peripheral Blood Mononuclear Cells of Patients With Sepsis and How to Remove It - A Technical Report.
Ficoll
MACS
PBMC (peripheral blood mononuclear cells)
Sepsis
cell isolation
centrifugation
density gradient
low-density granulocytes
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2021
2021
Historique:
received:
22
03
2021
accepted:
02
08
2021
entrez:
6
9
2021
pubmed:
7
9
2021
medline:
29
10
2021
Statut:
epublish
Résumé
Elucidating the mechanisms contributing to the dysregulated host response to infection as part of the syndrome is a current challenge in sepsis research. Peripheral blood mononuclear cells are widely used in immunological studies. Density gradient centrifugation, a common method, is of limited use for blood drawn from patients with sepsis. A significant number of low-density granulocytes co-purify contributing to low purity of isolated peripheral blood mononuclear cells. Whole blood anticoagulated with lithium heparin was drawn from patients with sepsis (n=14) and healthy volunteers (n=11). Immediately after drawing, the plasma fraction was removed and PBMC were isolated from the cellular fraction by density gradient centrifugation. Samples derived from patients with sepsis were subsequently incubated with cluster of differentiation 15 MicroBeads and granulocytes were depleted using magnetic-activated cell sorting. Core cellular functions as antigen presentation and cytokine secretion were analyzed in cells isolated from healthy volunteers (n=3) before and after depletion to confirm consistent functionality. We report here that depleting CD15
Identifiants
pubmed: 34484182
doi: 10.3389/fimmu.2021.684119
pmc: PMC8416421
doi:
Types de publication
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
684119Informations de copyright
Copyright © 2021 Schenz, Obermaier, Uhle, Weigand and Uhle.
Déclaration de conflit d'intérêts
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Références
PLoS One. 2014 Feb 13;9(2):e85696
pubmed: 24551035
JAMA. 2016 Feb 23;315(8):801-10
pubmed: 26903338
Am J Respir Crit Care Med. 2017 Feb 1;195(3):331-338
pubmed: 27513822
Cryobiology. 2009 Dec;59(3):366-8
pubmed: 19766618
J Immunol. 2010 Mar 15;184(6):3284-97
pubmed: 20164424
Shock. 2020 Sep;54(3):285-293
pubmed: 31764621
Intensive Care Med. 2008 May;34(5):912-6
pubmed: 18180900
J Trauma. 1988 Mar;28(3):353-61
pubmed: 3351993
Crit Care Med. 2018 Jun;46(6):915-925
pubmed: 29537985
PLoS One. 2012;7(11):e48939
pubmed: 23152825
J Leukoc Biol. 2020 May;107(5):809-818
pubmed: 32170882
Scand J Immunol. 2019 Apr;89(4):e12748
pubmed: 30667541
J Trauma. 1989 Mar;29(3):277-83
pubmed: 2784506
J Biol Chem. 2018 Sep 7;293(36):13874-13888
pubmed: 30006348
Shock. 2014 Nov;42(5):383-91
pubmed: 25051284
Crit Care. 2014 Aug 01;18(4):R163
pubmed: 25084831
Shock. 2019 Jan;51(1):97-104
pubmed: 29461465
Trends Mol Med. 2014 Apr;20(4):195-203
pubmed: 24581450
Crit Care. 2018 Sep 30;22(1):248
pubmed: 30268141
J Immunol. 2019 Oct 15;203(8):2088-2099
pubmed: 31501258
Crit Care Med. 2003 Apr;31(4):1250-6
pubmed: 12682500
Biomed Res Int. 2018 Mar 26;2018:7497314
pubmed: 29780830
Lancet Infect Dis. 2019 Dec;19(12):e422-e436
pubmed: 31630991
Mediators Inflamm. 2020 Jan 22;2020:4370983
pubmed: 32214905
Front Immunol. 2019 Oct 11;10:2358
pubmed: 31681271
Free Radic Biol Med. 2018 Apr;118:23-34
pubmed: 29471107
Clin Nutr. 2020 Mar;39(3):958-965
pubmed: 31005335
Front Immunol. 2018 Sep 05;9:1926
pubmed: 30233566
Lancet Respir Med. 2017 Oct;5(10):816-826
pubmed: 28864056
Semin Immunopathol. 2013 Jul;35(4):455-63
pubmed: 23553215
Biomed Res Int. 2020 Feb 28;2020:9152140
pubmed: 32258157
BMC Infect Dis. 2018 Jul 31;18(1):355
pubmed: 30064357
PLoS One. 2016 Apr 13;11(4):e0153567
pubmed: 27073889