Immune phenotyping in a pediatric multicenter transplant study: Suitability of a preformulated dry-antibody panel system

Flow-cytometric immune phenotyping is influenced by cryopreservation and inter-laboratory variability limiting comparability in multicenter studies. We assessed a system of optimized, pre-mixed dry-antibody panel tubes requiring small amounts of whole blood for validity, reliability and challenges i...

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Veröffentlicht in:Human immunology 2024-09, Vol.85 (5), p.110837, Article 110837
Hauptverfasser: Ionescu, Lavinia I., Blydt-Hansen, Tom, Foster, Bethany J., Allen, Upton, Birk, Patricia E., Hamiwka, Lorraine, Phan, Veronique, Min, Sandar, Ivison, Sabine, Levings, Megan, West, Lori J., Mital, Seema, Urschel, Simon
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Sprache:eng
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Zusammenfassung:Flow-cytometric immune phenotyping is influenced by cryopreservation and inter-laboratory variability limiting comparability in multicenter studies. We assessed a system of optimized, pre-mixed dry-antibody panel tubes requiring small amounts of whole blood for validity, reliability and challenges in a Canadian multicenter study (POSITIVE) with long-distance sample shipping, using standardized protocols. Thirty-seven children awaiting solid-organ transplant were enrolled for parallel immune-phenotyping with both validated, optimized in-house panels and the dry-antibody system. Samples were collected before, 3 and 12 months post-transplant. Quality-assurance measures and congruence of phenotypes were compared using Bland-Altman comparisons, linear regression and group comparisons. Samples showed excellent lymphocyte viability (mean 94.8 %) and recovery when processed within 30 h. Comparing staining methods, significant correlations (Spearman correlation coefficient >0.6, p 
ISSN:0198-8859
1879-1166
1879-1166
DOI:10.1016/j.humimm.2024.110837