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 |
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| creator | 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 |
| description | 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 |
| doi_str_mv | 10.1016/j.humimm.2024.110837 |
| format | Article |
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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.</description><identifier>ISSN: 0198-8859</identifier><identifier>ISSN: 1879-1166</identifier><identifier>EISSN: 1879-1166</identifier><identifier>DOI: 10.1016/j.humimm.2024.110837</identifier><identifier>PMID: 39013208</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Flow cytometry ; Immune phenotyping ; Multicenter study ; Pediatric transplantation</subject><ispartof>Human immunology, 2024-09, Vol.85 (5), p.110837, Article 110837</ispartof><rights>2024 American Society for Histocompatibility and Immunogenetics</rights><rights>Copyright © 2024 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c241t-57a5ebbe46afae1cc6bc1a26842fa63586f40334ca680a85009eeab897ae4bdf3</cites><orcidid>0000-0002-8864-0197 ; 0000-0002-5209-2993 ; 0000-0003-0544-3102 ; 0000-0002-2753-442X ; 0000-0001-7053-1796</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0198885924000971$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/39013208$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ionescu, Lavinia I.</creatorcontrib><creatorcontrib>Blydt-Hansen, Tom</creatorcontrib><creatorcontrib>Foster, Bethany J.</creatorcontrib><creatorcontrib>Allen, Upton</creatorcontrib><creatorcontrib>Birk, Patricia E.</creatorcontrib><creatorcontrib>Hamiwka, Lorraine</creatorcontrib><creatorcontrib>Phan, Veronique</creatorcontrib><creatorcontrib>Min, Sandar</creatorcontrib><creatorcontrib>Ivison, Sabine</creatorcontrib><creatorcontrib>Levings, Megan</creatorcontrib><creatorcontrib>West, Lori J.</creatorcontrib><creatorcontrib>Mital, Seema</creatorcontrib><creatorcontrib>Urschel, Simon</creatorcontrib><title>Immune phenotyping in a pediatric multicenter transplant study: Suitability of a preformulated dry-antibody panel system</title><title>Human immunology</title><addtitle>Hum Immunol</addtitle><description>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.</description><subject>Flow cytometry</subject><subject>Immune phenotyping</subject><subject>Multicenter study</subject><subject>Pediatric transplantation</subject><issn>0198-8859</issn><issn>1879-1166</issn><issn>1879-1166</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><recordid>eNp9kMFO3TAQRa2qqLxC_6CqvOwmr3biOA4LJISgRUJi0bK2Js6k-Cl2gu1Uzd_XTwGWXc3m3Lm6h5DPnO054_LbYf-0OOvcvmSl2HPOVNW8IzuumrbgXMr3ZMd4qwql6vaUfIzxwBhrWCM-kNOqZbwqmdqRv3fOLR7p_IR-Suts_W9qPQU6Y28hBWuoW8ZkDfqEgaYAPs4j-ERjWvr1gv5cbILOjjatdBqOwYDDFHIIEva0D2uRadtN_Upn8DjSuMaE7pycDDBG_PRyz8jj7c2v6x_F_cP3u-ur-8KUgqeibqDGrkMhYQDkxsjOcCilEuUAsqqVHASrKmFAKgaqZqxFhE61DaDo-qE6I1-3v3OYnheMSTsbDY55A05L1BVTvGm4qOuMig01YYoxz9BzsA7CqjnTR-f6oDfn-uhcb85z7MtLw9I57N9Cr5IzcLkBmHf-sRh0NBa9yYYDmqT7yf6_4R_aVJgx</recordid><startdate>20240901</startdate><enddate>20240901</enddate><creator>Ionescu, Lavinia I.</creator><creator>Blydt-Hansen, Tom</creator><creator>Foster, Bethany J.</creator><creator>Allen, Upton</creator><creator>Birk, Patricia E.</creator><creator>Hamiwka, Lorraine</creator><creator>Phan, Veronique</creator><creator>Min, Sandar</creator><creator>Ivison, Sabine</creator><creator>Levings, Megan</creator><creator>West, Lori J.</creator><creator>Mital, Seema</creator><creator>Urschel, Simon</creator><general>Elsevier Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-8864-0197</orcidid><orcidid>https://orcid.org/0000-0002-5209-2993</orcidid><orcidid>https://orcid.org/0000-0003-0544-3102</orcidid><orcidid>https://orcid.org/0000-0002-2753-442X</orcidid><orcidid>https://orcid.org/0000-0001-7053-1796</orcidid></search><sort><creationdate>20240901</creationdate><title>Immune phenotyping in a pediatric multicenter transplant study: Suitability of a preformulated dry-antibody panel system</title><author>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</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c241t-57a5ebbe46afae1cc6bc1a26842fa63586f40334ca680a85009eeab897ae4bdf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Flow cytometry</topic><topic>Immune phenotyping</topic><topic>Multicenter study</topic><topic>Pediatric transplantation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ionescu, Lavinia I.</creatorcontrib><creatorcontrib>Blydt-Hansen, Tom</creatorcontrib><creatorcontrib>Foster, Bethany J.</creatorcontrib><creatorcontrib>Allen, Upton</creatorcontrib><creatorcontrib>Birk, Patricia E.</creatorcontrib><creatorcontrib>Hamiwka, Lorraine</creatorcontrib><creatorcontrib>Phan, Veronique</creatorcontrib><creatorcontrib>Min, Sandar</creatorcontrib><creatorcontrib>Ivison, Sabine</creatorcontrib><creatorcontrib>Levings, Megan</creatorcontrib><creatorcontrib>West, Lori J.</creatorcontrib><creatorcontrib>Mital, Seema</creatorcontrib><creatorcontrib>Urschel, Simon</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Human immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ionescu, Lavinia I.</au><au>Blydt-Hansen, Tom</au><au>Foster, Bethany J.</au><au>Allen, Upton</au><au>Birk, Patricia E.</au><au>Hamiwka, Lorraine</au><au>Phan, Veronique</au><au>Min, Sandar</au><au>Ivison, Sabine</au><au>Levings, Megan</au><au>West, Lori J.</au><au>Mital, Seema</au><au>Urschel, Simon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immune phenotyping in a pediatric multicenter transplant study: Suitability of a preformulated dry-antibody panel system</atitle><jtitle>Human immunology</jtitle><addtitle>Hum Immunol</addtitle><date>2024-09-01</date><risdate>2024</risdate><volume>85</volume><issue>5</issue><spage>110837</spage><pages>110837-</pages><artnum>110837</artnum><issn>0198-8859</issn><issn>1879-1166</issn><eissn>1879-1166</eissn><abstract>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.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>39013208</pmid><doi>10.1016/j.humimm.2024.110837</doi><orcidid>https://orcid.org/0000-0002-8864-0197</orcidid><orcidid>https://orcid.org/0000-0002-5209-2993</orcidid><orcidid>https://orcid.org/0000-0003-0544-3102</orcidid><orcidid>https://orcid.org/0000-0002-2753-442X</orcidid><orcidid>https://orcid.org/0000-0001-7053-1796</orcidid></addata></record> |
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| source | Elsevier ScienceDirect Journals |
| subjects | Flow cytometry Immune phenotyping Multicenter study Pediatric transplantation |
| title | Immune phenotyping in a pediatric multicenter transplant study: Suitability of a preformulated dry-antibody panel system |
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