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2026 (English)In: Journal of Immunology, ISSN 0022-1767, E-ISSN 1550-6606, Vol. 215, no 6Article in journal (Refereed) Published
Abstract [en]
N-linked glycosylation (N-glyc) sites (N-X-S/T, X≠P) can be introduced by somatic hypermutation in immunoglobulin Fab regions. In patients with rheumatoid arthritis (RA), anti-citrullinated protein antibodies have a striking overrepresentation of Fab N-glycosylation. To further explore this, we sequenced B cell receptors (BCRs) from peripheral blood of 13 RA patients and 6 healthy control subjects, analyzing in total >250,000 heavy chain (VH) and >100,000 light chain sequences from both total B cells and citrullinated fibrinogen-reactive (Cit-Fib+) cells. Distribution of variable VH genes in VDJ DNA, and transcripts of unmutated IgM and class-switched BCR, revealed transcript gene-usage bias and higher VH4 in natural rearrangements by out-of-frame VDJ DNA in RA patients compared with control subjects. IgG Fab N-glyc sites were slightly more prominent in RA than control subjects (14.9% versus 12.1%; P = 0.048) with certain VH genes (e.g. VH1-18, VH1-69, VH3-9) displaying enriched N-glyc cumulative frequencies by somatic hypermutation. VH gene N-glyc hotspots were identified, explained by a lower threshold for codon conversion (i.e. K/S/T-X-S/T) and especially frequent in VH4s. Yet, RA patients had significantly more N-glyc in complementarity-determining region (CDR) 1 and 3 compared with control subjects. Expanded clonotypes with somatic hypermutation-induced N-glyc sites were delineated by network analysis in both the RA patients and control group, but patients with RA displayed more highly mutated class-switched members. Furthermore, Cit-Fib+ BCR-expanded clonotypes could be traced in the total B cell repertoire and exhibited increased frequency of N-glyc in mutated IgG/IgA subsets. Our findings highlight how Fab N-glyc sites can be linked to biased clonotype evolution and B cell selection in chronic responses
Place, publisher, year, edition, pages
Oxford University Press (OUP), 2026
Keywords
BCR repertoire, glycosylation, immunoglobulin, rheumatoid arthritis, V-gene sequencing
National Category
Immunology in the Medical Area
Identifiers
urn:nbn:se:ri:diva-81940 (URN)10.1093/jimmun/vkag113 (DOI)42269014 (PubMedID)2-s2.0-105042210463 (Scopus ID)
Note
QC 20260713
2026-07-132026-07-132026-07-13Bibliographically approved