Identification of proteinase 3 autoreactive CD4+T cells and their T-cell receptor repertoires in antineutrophil cytoplasmic antibody-associated vasculitis

Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is an autoimmune disease involving autoreactivity to proteinase 3 (PR3) as demonstrated by presence of ANCAs. While autoantibodies are screened for diagnosis, autoreactive T cells and their features are less well-studied. Here, we investigated PR3-specific CD4+T cell responses and features of autoreactive T cells in patients with PR3-AAV, using a cohort of 72 patients with either active or inactive disease. Autoreactive PR3-specific CD4+T cells producing interferon g in response to protein stimulation were found to express the G-protein coupled receptor 56 (GPR56), a cell surface marker that distinguishes T cells with cytotoxic capacity. GPR56+CD4+T cells were significantly more prominent in the blood of patients with inactive as compared to active disease, suggesting that these cells were affected by immunosuppression and/or that they migrated from the circulation to sites of organ involvement. Indeed, GPR56+CD4+T cells were identified in T-cell infiltrates of affected kidneys and an association with immunosuppressive therapy was found. Moreover, distinct TCR gene segment usage and shared (public) T cell clones were found for the PR3-reactive TCRs. Shared T cell clones were found in different patients with AAV carrying the disease-associated HLA- DP allele, demonstrating convergence of the autoreactive T cell repertoire. Thus, we identified a CD4+T cell signature in blood and in affected kidneys that display PR3 autoreactivity and associates with T cell cytotoxicity. Our data provide a basis for novel rationales for both immune monitoring and future therapeutic intervention in PR3-AAV.Funding Agencies|National Genomics Infrastructure in Stockholm - Science for Life Laboratory; Knut and Alice Wallenberg Foundation; Swedish Research Council; SNIC/Uppsala Multidisciplinary Center for Advanced Computational Science [2019-01664]; Karolinska Institutets Foundation for Rheumatology Research; Karolinska Institutets Research Foundation [2019-01961, 2020-02558]; Alex and Eva Wallstroem Foundation [2020-01647]; Tore Nilsons Foundations for Medical Research [2021-00096]; Stockholm County Council; Swedish Society for Medical Research [2019-01664, S17-0104]; ake Wiberg Foundation ( [20190859]; Magnus Bergvall Foundation [M19-0665]; [2019-03538]</p