THU0211 EVOLUTION OF KIDNEY ANTIBODY SECRETING CELLS MOLECULAR SIGNATURE IN LUPUS PATIENTS WITH ACTIVE NEPHRITIS UPON IMMUNOSUPPRESSIVE THERAPY

Abstract

Background: Pathogenic antibody secreting cells (ASC) have been identified in the kidney of SLE-prone mice, but are poorly characterized in human lupus nephritis (LN). We hypothesized that long-lived plasma cells may contribute to the failure of immunosuppressive therapy in refractory patients. Objectives: To characterize and compare the single cell molecular signature of ASC in kidney and urine from patients with active LN, either untreated or after immunosuppressive therapy failure. Methods: We included patients with biopsy proven active LN from 4 centers and meeting the ACR revised classification criteria for SLE diagnosis. Renal biopsies were scored according to 2003 International Society of Nephrology/Renal Pathology Society (ISN/RPS) classification, and stained with anti-CD138 to visualize ASC. ASC were single cell sorted as CD3-/CD14-/CD16-/CD27high/CD38high cells. Single-cell gene expression profiling was performed by multiplex RT-PCR using Fluidigm Dynamic Arrays. We used a set of genes derived from a previous transcriptomic analysis of human splenic and bone marrow ASC to distinguish the process of ASC maturation from plasmablast (PB) to long-lived PC. We also studied ASC transcriptional program from urine of untreated LN patients at diagnosis and after 3 and 6 months of a prospective follow up during induction therapy (Plasmo-Lup study). Results: Immunohistochemistry stainings on kidney biopsies from both untreated (N=15) and refractory patients (N=6) showed infiltrates of CD138+ ASC mainly located in the interstitium, particularly in untreated patients. Single cell molecular signature of kidney ASC from 3 untreated patients with class IV LN revealed that these cells were mostly PB expressing multiple genes linked with cell division, and PC without long-lived genes expression. This contrasted with ASC signature from 3 patients with active LN and mycophenolate mofetil (MMF) failure that expressed long-lived PC genes and no proliferative genes. Primary component analysis of 170 single-cells showed clustering of ASC from MMF treated patients with long-lived bone marrow PC from healthy donors that were distinct from PB/PC from untreated patients (Figure 1). A PB signature was observed in urine ASC at diagnosis, similar to their kidney counterpart. The concentration of ASC in urine in 22 untreated patients correlated with ISN/RPS classification, with higher concentration in class IV patients (p Conclusion: These results suggest that PB infiltrate kidney of untreated LN patients, and that kidney long-lived PC may contribute to the failure of immunosuppressive therapy. Acknowledgement: This work was supported by Foreum Disclosure of Interests: None declared