Improving naive B cell isolation by absence of CD45RB glycosylation and CD27 expression in combination with BCR isotype.


Journal

European journal of immunology
ISSN: 1521-4141
Titre abrégé: Eur J Immunol
Pays: Germany
ID NLM: 1273201

Informations de publication

Date de publication:
10 2022
Historique:
revised: 06 07 2022
received: 26 05 2022
accepted: 20 07 2022
pubmed: 22 7 2022
medline: 14 10 2022
entrez: 21 7 2022
Statut: ppublish

Résumé

In past years ex vivo and in vivo experimental approaches involving human naive B cells have proven fundamental for elucidation of mechanisms promoting B cell differentiation in both health and disease. For such studies, it is paramount that isolation strategies yield a population of bona fide naive B cells, i.e., B cells that are phenotypically and functionally naive, clonally non-expanded, and have non-mutated BCR variable regions. In this study different combinations of common as well as recently identified B cell markers were compared to isolate naive B cells from human peripheral blood. High-throughput BCR sequencing was performed to analyze levels of somatic hypermutation and clonal expansion. Additionally, contamination from mature mutated B cells intrinsic to each cell-sorting strategy was evaluated and how this impacts the purity of obtained populations. Our results show that current naive B cell isolation strategies harbor contamination from non-naive B cells, and use of CD27-IgD

Identifiants

pubmed: 35862268
doi: 10.1002/eji.202250013
doi:

Substances chimiques

Immunoglobulin D 0
Immunoglobulin Isotypes 0
Immunoglobulin M 0
Tumor Necrosis Factor Receptor Superfamily, Member 7 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1630-1639

Informations de copyright

© 2022 Wiley-VCH GmbH.

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Auteurs

Jana Koers (J)

Sanquin Research, Department of Immunopathology, and Landsteiner Laboratory, Amsterdam University medical centers, University of Amsterdam, Amsterdam, The Netherlands.

Sabrina Pollastro (S)

Sanquin Research, Department of Immunopathology, and Landsteiner Laboratory, Amsterdam University medical centers, University of Amsterdam, Amsterdam, The Netherlands.

Simon Tol (S)

Sanquin Research, Department of Research facilities, and Landsteiner Laboratory, Amsterdam University medical centers, University of Amsterdam, Amsterdam, The Netherlands.

Ilse T G Niewold (ITG)

Department of Rheumatology and Clinical Immunology, ARC, Amsterdam UMC, location AMC, University of Amsterdam, Amsterdam, The Netherlands.

Pauline A van Schouwenburg (PA)

Department of Pediatrics, Laboratory for Pediatric Immunology, Leiden University Medical Center, Leiden, Netherlands.

Niek de Vries (N)

Department of Rheumatology and Clinical Immunology, ARC, Amsterdam UMC, location AMC, University of Amsterdam, Amsterdam, The Netherlands.

Theo Rispens (T)

Sanquin Research, Department of Immunopathology, and Landsteiner Laboratory, Amsterdam University medical centers, University of Amsterdam, Amsterdam, The Netherlands.

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