Alloreactive memory B cell detection by flow cytometric cross match using polyclonally activated memory B cell culture supernatants

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dc.authoridATA, PINAR/0000-0002-6688-2347
dc.authorscopusid57216912962
dc.authorscopusid56769801000
dc.authorscopusid57384153600
dc.authorscopusid42360894600
dc.authorscopusid57729497100
dc.authorscopusid57729949000
dc.authorscopusid8624948000
dc.authorwosidATA, PINAR/AAC-9967-2020
dc.contributor.authorAkalan, Hande
dc.contributor.authorŞirin, Duygu Yaşar
dc.contributor.authorYılmaz, İpek
dc.contributor.authorAta, Pınar
dc.contributor.authorKara, Veli Melih
dc.contributor.authorTaşdemir, Nicel
dc.contributor.authorBilgen, Türker
dc.contributor.authorTitiz, Mesut İzzet
dc.date.accessioned2023-04-20T08:01:12Z
dc.date.available2023-04-20T08:01:12Z
dc.date.issued2022
dc.departmentFakülteler, Fen Edebiyat Fakültesi, Biyoloji Bölümü
dc.departmentYüksekokullar, Sağlık Yüksekokulu, Beslenme ve Diyetetik Bölümü
dc.departmentRektörlüğe Bağlı Bölümler, Rektörlük, Bilimsel ve Teknolojik Araştırmalar Uygulama ve Araştırma Merkezi
dc.description.abstractIn addition to alloantibodies, alloreactive memory B cell (mBC) evaluation has a potential for immunological risk assessment during transplantation processes. For the alloreactive mBCs evaluation currently, direct Flow Cytometric (FC) analysis using the HLA tetramer staining is an option. Evaluation of alloantibodies produced by the polyclonally stimulated alloreactive mBCs in in vitro culture system seems to be another useful approach, but this needs further downstream applications. In this study, we investigated the usefulness of the Flow Cytometric Cross Match (FCXM-supernatant) in which in vitro polyclonally activated mBCs culture supernatants and potential donor's lymphocytes being used for the mBC detection. FCXM-supernatant assays were performed between culture supernatants of polyclonally activated mBCs obtained from 4 allosensitized multiparous women and 14 renal transplant patients, and their non-alloimmunized spouses' or donors' lymphocytes, and vice versa. HLA typing was performed by SSP method. Anti-HLA antibodies produced by in vitro activated alloreactive mBCs were also evaluated by the Luminex assays. The success of in vitro polyclonal activation of mBCs was evaluated by a total IgG ELISA test and antibody secreting cell analyses by FC. Donor specific alloreactive mBCs were detected by FCXM-supernatant in 45% of the 18 allosensitized cases. Detection rate was 85% (6 out of 7) in the strongly allosensitized cases. No alloreactive mBCs was detected in control cases without allosensitization. FCXMsupernatant negative results of the allosensitized cases were related to low level of allosensitization and insufficient polyclonal stimulation evaluated by total IgG antibody tests of the supernatants. We herein report a practical methodology for alloreactive mBC detection as a donor specific manner using the FCXM-supernatant assay so that this would easily be transformed into a routine test performed in tissue typing laboratories.
dc.description.sponsorshipTUBITAK; The Scientific and Technological Research Council of Turkey [217S630]; Research Fund of the Tekirdag Namik Kemal University [NKUBAP.01.GA.21.301]
dc.description.sponsorshipThis study was supported by the TUBITAK; The Scientific and Technological Research Council of Turkey (Grant number: 217S630) and by the Research Fund of the Tekirdag Namik Kemal University (Project Number: NKUBAP.01.GA.21.301). We would like to thank Drs. Frans H.J. Claas and Gonca E. Karahan from Leiden University Medical Center, and Dr. Fahri Ucar from Akdeniz University Tissue Typing Laboratory for their supports.
dc.identifier.doi10.1016/j.trim.2022.101642
dc.identifier.issn0966-3274
dc.identifier.issn1878-5492
dc.identifier.pmid35667546
dc.identifier.scopus2-s2.0-85131407806
dc.identifier.scopusqualityQ3
dc.identifier.urihttps://doi.org/10.1016/j.trim.2022.101642
dc.identifier.urihttps://hdl.handle.net/20.500.11776/10786
dc.identifier.volume73
dc.identifier.wosWOS:000812972700003
dc.identifier.wosqualityQ4
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorAkalan, Hande
dc.institutionauthorŞirin, Duygu Yaşar
dc.institutionauthorKara, Veli Melih
dc.institutionauthorBilgen, Türker
dc.institutionauthorTitiz, Mesut İzzet
dc.language.isoen
dc.publisherElsevier
dc.relation.ispartofTransplant Immunology
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectFlow Cytometric Cross Match
dc.subjectAlloreactive Memory B Cell
dc.subjectCulture Supernatants
dc.subjectPolyclonal Activation
dc.subjectHumoral Rejection
dc.subjectHla
dc.subjectQuantification
dc.titleAlloreactive memory B cell detection by flow cytometric cross match using polyclonally activated memory B cell culture supernatants
dc.typeArticle

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Bilimsel ve Teknolojik Araştırmalar Uygulama ve Araştırma Merkezi Koleksiyonu
Fen Edebiyat Fakültesi Koleksiyonu
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