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

Autor: Ionescu LI; Division of Pediatric Cardiology, University of Alberta, Edmonton, Alberta, Canada; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada., Blydt-Hansen T; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Division of Pediatric Nephrology, University of British Columbia, Vancouver, Canada., Foster BJ; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Division of Nephrology, Montreal Children's Hospital, McGill University Health Centre, Montreal, Quebec, Canada., Allen U; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Division of Infectious Diseases, Hospital for Sick Children, Toronto, Ontario, Canada., Birk PE; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Department of Pediatrics and Child Health, Health Sciences Centre Winnipeg, Winnipeg, Manitoba, Canada., Hamiwka L; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Division of Nephrology, Alberta Children's Hospital, University of Calgary, Calgary, Canada., Phan V; Division of Nephrology, Department of Pediatrics, Centre Hospitalier Universitaire Sainte-Justine, Montreal, Quebec, Canada., Min S; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Genetics and Genome Biology Program, Hospital for Sick Children, Toronto, Ontario, Canada., Ivison S; University of British Columbia, Vancouver, Canada., Levings M; University of British Columbia, Vancouver, Canada., West LJ; Division of Pediatric Cardiology, University of Alberta, Edmonton, Alberta, Canada; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada., Mital S; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada; Genetics and Genome Biology Program, Hospital for Sick Children, Toronto, Ontario, Canada; Division of Cardiology, Department of Pediatrics, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada., Urschel S; Division of Pediatric Cardiology, University of Alberta, Edmonton, Alberta, Canada; Canadian Donation and Transplant Research Program, Edmonton, Alberta, Canada. Electronic address: urschel@ualberta.ca.
Jazyk: angličtina
Zdroj: Human immunology [Hum Immunol] 2024 Sep; Vol. 85 (5), pp. 110837. Date of Electronic Publication: 2024 Jul 15.
DOI: 10.1016/j.humimm.2024.110837
Abstrakt: 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 < 0.05), mean difference <5 % and variation 2SD <25 % were found for natural-killer, T and B cells, including many immunologically important cell subsets (CD8+, naïve, memory CD4+ T; switched-memory, transitional B). Some subgroups (plasmablasts, CD1d+CD5hi B cells) showed weak correlations, limiting interpretation reliability. The dry-antibody system provides a reliable method for standardized analysis of many immune phenotypes after long-distance shipping when processed within 30 h, rendering the system attractive for pediatric studies due to small blood amounts required and highly standardized processing and analysis.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Seema Mital reports a relationship with Bristol Myers Squibb that includes: consulting or advisory. Seema Mital reports a relationship with Tenaya Therapeutics that includes: consulting or advisory. Lavinia Ionescu reports a relationship with Beckman Coulter Canada that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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