Improving naive B cell isolation by absence of CD45RB glycosylation and CD27 expression in combination with BCR isotype. Issue 10 (16th August 2022)

• Record Type: Journal Article
• Authors: Koers, Jana; Pollastro, Sabrina; Tol, Simon; Niewold, Ilse T.G.; van Schouwenburg, Pauline A.; de Vries, Niek; Rispens, Theo
• Is Part Of: European journal of immunology. Volume 52:Issue 10(2022)
• Page Start: 1630
• Page End: 1639
• Publication Date: 2022-08-16
• Subjects: B cell receptor -- CD27 -- CD45RB glycosylation -- Naive B cells -- somatic hypermutation; Immunology -- Periodicals; 616.079
• DOI: 10.1002/eji.202250013
• Languages: English
• ISSNs: 0014-2980

Abstract

Abstract: 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 + is adequate but can be improved by including markers for CD45RB glycosylation and IgM. The finetuning of naive B cell classification provided herein will harmonize research lines using naive B cells, and will improve B cell profiling during health and disease, e.g. during diagnosis, treatment, and vaccination strategies. Abstract : B cell‐sorting strategy using markers for a novel glycosylation‐dependent epitope on CD45RB, CD27 and B cell receptor isotype improve isolation of human naive B cells and yield a naive B cell population with limited IGHV somatic hypermutations and clonal expansion.