AB0021 CHARACTERISATION OF AGE-ASSOCIATED B CELLS IN EARLY, DRUG NAÏVE RHEUMATOID ARTHRITIS

Abstract

Background:Rheumatoid arthritis (RA) is a chronic inflammatory disease of immune dysregulation affecting the joints. While T cells play a recognised role in disease pathogenesis, the presence of autoantibodies years before the clinical onset of disease, and the efficacy of the B cell-depleting therapy rituximab, highlight a pathogenic role for these functionally diverse lymphocytes. A novel subset, termed age-associated B cells (ABCs), are described as CD19high CD21- CD11c+, with a high proportion expressing the transcription factor T-bet; they are elevated in murine models of autoimmunity and produce autoantibodies characteristic of autoimmune disease. A detailed characterisation of ABCs in early, drug naïve RA has not yet been conducted.Objectives:We aimed to characterise peripheral blood (PB) and synovial fluid (SF) ABCs in patients suffering from early drug naïve RA. As a secondary objective we sought to determine whether this population differs between RA patients and age-matched early psoriatic arthritis (PsA; disease controls) and healthy controls.Methods:Newly presenting early RA and other inflammatory arthritis patients, naïve to immunomodulatory treatment, were recruited from the Newcastle Early Arthritis Clinic. Patients with established RA (≥1 year duration on treatment and age-matched healthy controls were recruited in parallel. B cell subsets (naïve (CD19+IgD+CD27-), switched memory (CD19+IgD-CD27+) and ABC (CD19+CD21-CD11c+)) in PB and SF were characterised by flow cytometry. FACS sorted PB B cell subset gene expression profiles were assessed using a customised NanoString nCounter Human Immunology v2 Panel.Results:Transcriptionally, ABC differed from both naïve and switched memory B cells. In keeping with published studies ABCs had an activated memory phenotype, exemplified by elevated expression of CD69, CD80, and CD86, as well as Ki67, HLA-DR, IgG and T-bet. Interestingly, ABC had high expression of the Fc Receptor Like (FcRL) family (FcRL1-5), and an inflammatory homing profile (high levels of CXCR3 and low levels of CXCR4 and CXCR5). We found no difference in the proportion of PB ABC between RA patients and control groups, and no association of ABC frequencies with age. Focussing on RA ABCs specifically, we observed elevated ABC frequencies in females, but no association with disease activity. In keeping with their chemokine receptor profile, cross-sectional analysis also demonstrated ABC enrichment in SF compared to PB, with SF frequencies higher in established than early disease. The transcriptome of ABCs from early RA patients differed from both age-matched disease controls and healthy donors (Figure 1 next page).Conclusion:These data demonstrate that ABCs have a unique, activated, class-switched proliferative phenotype that is transcriptionally distinct from switched memory and naïve B cells. Interestingly, at the transcriptome level early RA ABCs differ to their counterparts in health and other inflammatory arthritides, suggesting they may differ functionally and contribute to pathogenesis. Of note, their high expression of MHC class II (HLA-DR) and co-stimulatory (CD80 and CD86) molecules suggests an important antigen-presentation function of ABC, which together with their unique FcRL family expression pattern, warrants further functional characterisation.Figure 1.Differential gene expression in ABCs from early RA patients compared to early PsA patients (A) and healthy controls (B). A NanoString nCounter Technologies chip was used to assess gene expression. Raw counts were normalised to the housekeeping genes. Sample quality was then assessed using the arrayQualityMetrics package. Gene expression profiles between different donor groups was assessed using the DESeq2 R package. In both hierarchical clustering heatmaps gene expression intensities were log2 transformed and their z-scores are displayed as colours ranging from yellow (low expression) to red (high expression) as shown in the key.Disclosure of Interests:Gemma Vidal-Pedrola: None declared, Najib Naamane: None declared, Dagmar Scheel-Toellner: None declared, Arthur Pratt Grant/research support from: GSK, Pfizer and Janssen, Andrew Mellor: None declared, John D Isaacs Speakers bureau: Abbvie, Gilead, Roche, UC, Consultant of: Abbvie, Gilead, Roche, UC, Grant/research support from: Pfizer, GSK and Janssen, Amy E. Anderson Grant/research support from: Pfizer, GSK and Janssen