Circulating autoreactive proteinase 3+ B cells and tolerance checkpoints in ANCA-associated vasculitis

Alvise Berti; Sophie Hillion; Amber M Hummel; Young Min Son; Nedra Chriti; Tobias Peikert; Eva M Carmona; Wayel H Abdulahad; Peter Heeringa; Kristina M Harris; E William St Clair; Paul Brunetta; Fernando C Fervenza; Carol A Langford; Cees Gm Kallenberg; Peter A Merkel; Paul A Monach; Philip Seo; Robert F Spiera; John H Stone; Guido Grandi; Jie Sun; Jacques-Olivier Pers; Ulrich Specks; Divi Cornec; RAVE-ITN Research Group

doi:10.1172/jci.insight.150999

Circulating autoreactive proteinase 3+ B cells and tolerance checkpoints in ANCA-associated vasculitis

JCI Insight. 2021 Nov 22;6(22):e150999. doi: 10.1172/jci.insight.150999.

Authors

Alvise Berti^{1

2}, Sophie Hillion³, Amber M Hummel¹, Young Min Son^{1

4}, Nedra Chriti³, Tobias Peikert¹, Eva M Carmona^{1

4}, Wayel H Abdulahad⁵, Peter Heeringa⁵, Kristina M Harris⁶, E William St Clair⁷, Paul Brunetta⁸, Fernando C Fervenza⁹, Carol A Langford¹⁰, Cees Gm Kallenberg⁵, Peter A Merkel¹¹, Paul A Monach¹², Philip Seo¹³, Robert F Spiera¹⁴, John H Stone¹⁵, Guido Grandi², Jie Sun^{1

4}, Jacques-Olivier Pers³, Ulrich Specks¹, Divi Cornec^{1

3}; RAVE-ITN Research Group

Affiliations

¹ Thoracic Disease Research Unit, Division of Pulmonary and Critical Care Medicine, Mayo Clinic, Rochester, Minnesota, USA.
² Rheumatology Unit, S. Chiara Regional Hospital and Department of CIBIO, University of Trento, Trento, Italy.
³ INSERM UMR1227, Lymphocytes B et Autoimmunité, University of Brest, CHRU Brest, Brest, France.
⁴ Department of Immunology, Mayo Clinic, Rochester, Minnesota, USA.
⁵ Department of Rheumatology and Clinical Immunology and Department of Pathology and Medical Biology, University of Groningen, Groningen, Netherlands.
⁶ Immune Tolerance Network, Bethesda, Maryland, USA.
⁷ Division of Rheumatology, Duke University Medical Center, Durham, North Carolina, USA.
⁸ Genentech Inc., South San Francisco, California, USA.
⁹ Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, USA.
¹⁰ Center for Vasculitis Care and Research, Department of Rheumatic and Immunologic Diseases, Cleveland Clinic, Cleveland, Ohio, USA.
¹¹ Division of Rheumatology, Department of Medicine, Division of Clinical Epidemiology, Department of Biostatistics, Epidemiology, and Informatics, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
¹² Brigham and Women's Hospital and VA Boston Healthcare System, Boston Massachusetts, USA.
¹³ Division of Rheumatology, Johns Hopkins University, Baltimore, Maryland, USA.
¹⁴ Vasculitis & Scleroderma Program, Hospital for Special Surgery, New York, New York, USA.
¹⁵ Massachusetts General Hospital, Boston, Massachusetts, USA.

Abstract

BACKGROUNDLittle is known about the autoreactive B cells in antineutrophil cytoplasmic antibody-associated (ANCA-associated) vasculitis (AAV). We aimed to investigate tolerance checkpoints of circulating antigen-specific proteinase 3-reactive (PR3+) B cells.METHODSMulticolor flow cytometry in combination with bioinformatics and functional in vitro studies were performed on baseline samples of PBMCs from 154 well-characterized participants of the RAVE trial (NCT00104299) with severely active PR3-AAV and myeloperoxidase-AAV (MPO-AAV) and 27 healthy controls (HCs). Clinical data and outcomes from the trial were correlated with PR3+ B cells (total and subsets).RESULTSThe frequency of PR3+ B cells among circulating B cells was higher in participants with PR3-AAV (4.77% median [IQR, 3.98%-6.01%]) than in participants with MPO-AAV (3.16% median [IQR, 2.51%-5.22%]) and participants with AAV compared with HCs (1.67% median [IQR, 1.27%-2.16%], P < 0.001 for all comparisons), implying a defective central tolerance checkpoint in patients with AAV. Only PBMCs from participants with PR3-AAV contained PR3+ B cells capable of secreting PR3-ANCA IgG in vitro, proving they were functionally distinct from those of participants with MPO-AAV and HCs. Unsupervised clustering identified subtle subsets of atypical autoreactive PR3+ memory B cells accumulating through the maturation process in patients with PR3-AAV. PR3+ B cells were enriched in the memory B cell compartment of participants with PR3-AAV and were associated with higher serum CXCL13 levels, suggesting an increased germinal center activity. PR3+ B cells correlated with systemic inflammation (C-reactive protein and erythrocyte sedimentation rate, P < 0.05) and complete remission (P < 0.001).CONCLUSIONThis study suggests the presence of defective central antigen-independent and peripheral antigen-dependent checkpoints in patients with PR3-AAV, elucidating the selection process of autoreactive B cells.Trial registrationClinicalTrials.gov NCT00104299.FundingThe Vasculitis Foundation, the National Institute of Allergy and Infectious Diseases of the NIH, and the Mayo Foundation for Education and Research.

Keywords: Autoimmune diseases; Autoimmunity; Immunology; Vasculitis.

Publication types

Multicenter Study
Randomized Controlled Trial
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis / metabolism*
Double-Blind Method
Female
Flow Cytometry / methods*
Humans
Male
Memory B Cells / metabolism*
Peptide Hydrolases / metabolism*

Circulating autoreactive proteinase 3+ B cells and tolerance checkpoints in ANCA-associated vasculitis

Authors

Affiliations

Abstract

Publication types

MeSH terms

Substances

Associated data

Grants and funding