Assessment of IgG-Fc glycosylation from individual RhD-specific B cell clones reveals regulation at clonal rather than clonotypic level

Erik L. de Graaf, Mads Delbo Larsen, Nieke van der Bolt, Remco Visser, Onno J.H.M. Verhagen, Agnes L. Hipgrave Ederveen, Carolien A.M. Koeleman, C. Ellen van der Schoot, Manfred Wuhrer, Gestur Vidarsson*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The type and strength of effector functions mediated by immunoglobulin G (IgG) antibodies rely on the subclass and the composition of the N297 glycan. Glycosylation analysis of both bulk and antigen-specific human IgG has revealed a marked diversity of the glycosylation signatures, including highly dynamic patterns as well as long-term stability of profiles, yet information on how individual B cell clones would contribute to this diversity has hitherto been lacking. Here, we assessed whether clonally related B cells share N297 glycosylation patterns of their secreted IgG. We differentiated single antigen-specific peripheral IgG+ memory B cells into antibody-secreting cells and analysed Fc glycosylation of secreted IgG. Furthermore, we sequenced the variable region of their heavy chain, which allowed the grouping of the clones into clonotypes. We found highly diverse glycosylation patterns of culture-derived IgG, which, to some degree, mimicked the glycosylation of plasma IgG. Each B cell clone secreted IgG with a mixture of different Fc glycosylation patterns. The majority of clones produced fully fucosylated IgG. B cells producing afucosylated IgG were scattered across different clonotypes. In contrast, the remaining glycosylation traits were, in general, more uniform. These results indicate IgG-Fc fucosylation to be regulated at the single-clone level, whereas the regulation of other glycosylation traits most likely occurs at a clonotypic or systemic level. The discrepancies between plasma IgG and culture-derived IgG, could be caused by the origin of the B cells analysed, clonal dominance or factors from the culture system, which need to be addressed in future studies.

Original languageEnglish
Pages (from-to)428-439
Number of pages12
JournalImmunology
Volume171
Issue number3
Early online date14 Dec 2023
DOIs
Publication statusPublished - Mar 2024

Keywords

  • antibodies
  • B cell
  • B cell receptor
  • cell differentiation
  • repertoire development

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