Isolation and characterisation of PR3-specific B cells and their immunoglobulin sequences

• Published in: Journal of autoimmunity 2024-01, Vol.142, p.103129-103129, Article 103129
• Main Authors: Kelly, Shane, Jackson, Katherine JL, Peters, Timothy J., Suan, Dan, Goodnow, Christopher C.
• Format: Article
• Language: English
• Subjects: Granulomatosus with polyangiitis; Immunoglobulin; Proteinase 3; Vasculitis
• ISSN: 0896-8411; 1095-9157
• DOI: 10.1016/j.jaut.2023.103129

PR3 autoantibodies are essential to the diagnosis and monitoring of granulomatosus with polyangiitis, but to date no PR3 autoantibody sequences have been published. To identify and characterise PR3-specific B cells from the peripheral blood of patients with PR3 autoantibodies. Peripheral blood mononuclear cells from seven patients with PR3 autoantibodies were stained with PR3. B cells that bound PR3 underwent single cell sorting, transcriptome sequencing, and their immunoglobulin sequences expressed as antibodies and tested for PR3-specificity by ELISA. We identified 19 PR3-specific B cells from only one PR3-seropositive patient at a low frequency (0.0075 % of B cells) in the peripheral blood. These were polyclonal, IgG+ and enriched for IgG4, lambda pairing, IGHJ6 gene usage, CDRH3 length, IGHE and CD71 expression. They demonstrated relatively low levels of somatic hypermutation and variably reduced PR3 binding when reverted to germline. Identifying PR3-specific B cells in the peripheral blood is possible but challenging and those we did identify exhibited features suggesting that PR3-self reactivity may occur early in B-cell development. •PR3-specific B cells may be identified from the blood of patients with PR3 autoantibodies, but at an extremely low frequency.•We report the immunoglobulin sequences from 19 PR3-specific B cells from a single patient.•PR3-specific B cells were enriched for IgG4, lambda pairing, IGHJ6 gene usage, CDRH3 length and IGHE gene expression.•PR3-antibody sequences showed low levels of somatic hypermutation & some germline-reverted sequences retained PR3 binding.•This suggests that PR3-self reactivity may occur early in B-cell development.