Towards standardizing basophil identification by flow cytometry.
CBC (complete blood count)
basophil actication test
basophil activating test (BAT)
basophil identification markers
flow cytometry
Journal
Frontiers in allergy
ISSN: 2673-6101
Titre abrégé: Front Allergy
Pays: Switzerland
ID NLM: 9918227355906676
Informations de publication
Date de publication:
2023
2023
Historique:
received:
28
12
2022
accepted:
27
01
2023
entrez:
20
3
2023
pubmed:
21
3
2023
medline:
21
3
2023
Statut:
epublish
Résumé
Basophils normally make up <2% of the white blood cells (WBC). There is no clear consensus for basophil identification by flow cytometry. The increased demand for basophil activation test (BAT) to identifying and monitoring allergic patients highlights the need for a standardized approach to identify basophils. Using flow cytometry we analyzed whole blood stained with antibodies against: IgE, CD123, CD193, CD203c, CD3, HLADR, FcɛRI, CRTH2 and CD45. We examined unstimulated blood as well as blood stimulated with Anti-IgE and fMLP. Finally, we compared the results to a complete blood count (CBC) from an FDA approved hematological analyzer. Basophil identification relying on just one surface marker performed worse than approaches utilizing two identification markers. The percentage of basophils from WBC determined by flow cytometry results had a good correlation with the CBC results even though the CBC results were generally higher. Stimulating whole blood with the basophil activators did not interfere with the basophil identification markers. In flow cytometry assays, two surface markers should be used for identifying basophils and if a very pure basophil fraction is desired a third marker can be considered. In our hands the approaches that included CD123 in combination with either CD193, HLADR
Sections du résumé
Background
UNASSIGNED
Basophils normally make up <2% of the white blood cells (WBC). There is no clear consensus for basophil identification by flow cytometry. The increased demand for basophil activation test (BAT) to identifying and monitoring allergic patients highlights the need for a standardized approach to identify basophils.
Methods
UNASSIGNED
Using flow cytometry we analyzed whole blood stained with antibodies against: IgE, CD123, CD193, CD203c, CD3, HLADR, FcɛRI, CRTH2 and CD45. We examined unstimulated blood as well as blood stimulated with Anti-IgE and fMLP. Finally, we compared the results to a complete blood count (CBC) from an FDA approved hematological analyzer.
Results
UNASSIGNED
Basophil identification relying on just one surface marker performed worse than approaches utilizing two identification markers. The percentage of basophils from WBC determined by flow cytometry results had a good correlation with the CBC results even though the CBC results were generally higher. Stimulating whole blood with the basophil activators did not interfere with the basophil identification markers.
Conclusion
UNASSIGNED
In flow cytometry assays, two surface markers should be used for identifying basophils and if a very pure basophil fraction is desired a third marker can be considered. In our hands the approaches that included CD123 in combination with either CD193, HLADR
Identifiants
pubmed: 36938328
doi: 10.3389/falgy.2023.1133378
pmc: PMC10020589
doi:
Types de publication
Journal Article
Langues
eng
Pagination
1133378Informations de copyright
© 2023 Sonder, Plassmeyer, Loizou and Alpan.
Déclaration de conflit d'intérêts
Author SUS, MP and DL are employed by Amerimmune. OA is the owner of Amerimmune. This study was funded by resources internally at Amerimmune. The funder (OA) had the following involvement with the study: Approved study design and the final manuscript. All authors declare no other competing interests.
Références
Clin Mol Allergy. 2005 Jun 30;3:9
pubmed: 15989690
Cytometry A. 2011 Feb;79(2):102-6
pubmed: 21265004
Clin Exp Allergy. 2017 Sep;47(9):1115-1124
pubmed: 28618090
J Allergy Clin Immunol. 1991 Sep;88(3 Pt 1):328-38
pubmed: 1716273
Curr Allergy Asthma Rep. 2018 Nov 15;18(12):77
pubmed: 30430289
Cytometry B Clin Cytom. 2020 Nov;98(6):483-490
pubmed: 32012452
J Allergy Clin Immunol. 2002 Jul;110(1):102-9
pubmed: 12110828
Biochem Med (Zagreb). 2015 Jun 05;25(2):141-51
pubmed: 26110027
Ann Biol Clin (Paris). 2012 Mar-Apr;70(2):141-54
pubmed: 22484525
Cytometry B Clin Cytom. 2015 May-Jun;88(3):183-9
pubmed: 25399741
Lancet. 1986 Feb 8;1(8476):307-10
pubmed: 2868172
Int J Lab Hematol. 2020 Jun;42(3):237-245
pubmed: 31841278
Cytometry. 1993;14(3):344-8
pubmed: 8472612
Int Arch Allergy Immunol. 2004 Apr;133(4):317-29
pubmed: 15031605
Allergy. 2011 Jan;66(1):85-91
pubmed: 20608915
Am J Clin Pathol. 2013 Sep;140(3):293-300
pubmed: 23955446
FEBS Lett. 1999 Oct 8;459(2):195-9
pubmed: 10518017
Clin Mol Allergy. 2008 Oct 16;6:12
pubmed: 18925959
Clin Transl Allergy. 2016 Apr 01;6:11
pubmed: 27042292
Clin Exp Allergy. 2008 Jun;38(6):921-8
pubmed: 18331364
J Investig Allergol Clin Immunol. 2010;20(6):535-6
pubmed: 21243941
Clin Exp Allergy. 2010 Jun;40(6):953; author reply 954
pubmed: 20557552
Allergy. 2021 Dec;76(12):3776-3788
pubmed: 33973252
J Allergy Clin Immunol. 2017 Mar;139(3):889-899.e11
pubmed: 27527263
Curr Allergy Asthma Rep. 2016 Jul;16(8):56
pubmed: 27411319
Front Immunol. 2016 Jul 07;7:260
pubmed: 27456009
Allergy. 2005 Nov;60(11):1446-50
pubmed: 16197480
Allergy. 2015 Nov;70(11):1393-405
pubmed: 26198455
Ann Lab Med. 2016 Jan;36(1):28-35
pubmed: 26522756
J Allergy Clin Immunol. 2010 Feb;125(2 Suppl 2):S73-80
pubmed: 20176269
Clin Exp Allergy. 2008 Jul;38(7):1239-40
pubmed: 18477015
Cytometry A. 2014 May;85(5):382-5
pubmed: 24753185
Cytometry B Clin Cytom. 2018 Sep;94(5):671-679
pubmed: 28493330
Cytometry A. 2014 Jul;85(7):570-2
pubmed: 24757016
Cytometry B Clin Cytom. 2021 Nov;100(6):632-644
pubmed: 33539657
Am J Clin Pathol. 2012 Jan;137(1):86-92
pubmed: 22180481
Allergy. 2009 Sep;64(9):1319-26
pubmed: 19243362
Clin Exp Allergy. 2010 Mar;40(3):411-8
pubmed: 20082620
Allergy. 2021 Aug;76(8):2420-2432
pubmed: 33475181