Optimized flow cytometric protocol for the detection of functional subsets of low frequency antigen-specific CD4
Antigen-specific cell detection
Cell enrichment
Cytokine response
Flow cytometry
Intracellular staining
Rare cell detection
Journal
MethodsX
ISSN: 2215-0161
Titre abrégé: MethodsX
Pays: Netherlands
ID NLM: 101639829
Informations de publication
Date de publication:
2020
2020
Historique:
received:
02
04
2020
accepted:
17
07
2020
entrez:
11
8
2020
pubmed:
11
8
2020
medline:
11
8
2020
Statut:
epublish
Résumé
Detection of low-frequency cells using flow cytometry is challenging, as the sensitivity of the analysis is dependent on the signal-to-noise ratio, and a cell frequency of 1 in 10,000 cells is accepted as the lower limit of detection for standard flow cytometry. A solution to this problem is to pre-enrich rare cell populations using magnetic-bead conjugated antibodies targeting lineage or activation markers. For measuring vaccine or pathogen induced immune responses, this method drastically increases the signal-to-noise ratio by enriching only activated (i.e., antigen-specific) cells and excluding all other peripheral blood leukocytes from the subsequent analysis. To date, magnetic enrichment of antigen-specific cells has only been described for CD4
Identifiants
pubmed: 32775228
doi: 10.1016/j.mex.2020.101005
pii: S2215-0161(20)30225-9
pii: 101005
pmc: PMC7396823
doi:
Types de publication
Journal Article
Langues
eng
Pagination
101005Informations de copyright
© 2020 The Author(s). Published by Elsevier B.V.
Déclaration de conflit d'intérêts
The authors declare that they have no conflict of interest.
Références
J Immunol Methods. 2019 Apr;467:48-57
pubmed: 30802449
Blood. 2013 Jul 18;122(3):405-12
pubmed: 23719298
J Exp Med. 1997 Jun 16;185(12):2089-94
pubmed: 9182680
Sci Rep. 2016 Jun 21;6:27944
pubmed: 27323685
Nat Med. 2005 Oct;11(10):1113-7
pubmed: 16186817
J Immunol. 2013 Apr 15;190(8):3967-76
pubmed: 23479226
Blood. 2007 Jul 1;110(1):201-10
pubmed: 17371945
Cytometry. 1990;11(2):231-8
pubmed: 1690625
Nat Med. 2005 Oct;11(10):1118-24
pubmed: 16186818
Clin Diagn Lab Immunol. 1996 May;3(3):301-4
pubmed: 8705673
Eur J Immunol. 2007 Sep;37(9):2370-7
pubmed: 17705136
Methods Mol Biol. 2018;1678:37-47
pubmed: 29071674
J Exp Med. 1998 Dec 21;188(12):2335-42
pubmed: 9858519