Single-cell transcriptomics reveal potent extrafollicular B cell response linked with granzyme K+ CD8 T cell activation in lupus kidney

Abstract

<h3>Objectives</h3> B and T cells constitute the majority of infiltrating lymphocytes in the kidney and represent the local perpetrators in lupus nephritis (LN), but the underlying pathogenic mechanisms are not well elucidated. The aim of this study is to explore the kidney-specific adaptive immune landscape in patients with active LN at the single-cell level. <h3>Methods</h3> We performed single-cell RNA/B cell receptor (BCR)/T cell receptor (TCR) sequencing analysis on sorting-purified B and T cells from the kidney and paired peripheral blood of patients with active LN, and the periphery of matched controls. Flow cytometry, Assay for Transposase Accessible-sequencing, multiplexed immunohistochemistry and functional studies were performed to validate the transcriptomic results. <h3>Results</h3> High infiltrations of intrarenal atypical B cells (ABCs) and antibody-secreting cells (ASCs) were identified in the B cell compartment. The single-cell BCR repertoire analysis revealed strong clonal expansion of intrarenal ASCs dominated by <i>IGHG1</i> and <i>IGHG3</i> isotypes, accompanied by lower frequencies of heavy-chain and light-chain somatic mutations, compared with the peripheral ASCs. Notably, a unique expansion of <i>IGHG4-59</i> and clonal overlap between ABCs and ASCs was found in kidney-specific clonotypes. In the T cell compartment, we identified granzyme K (GZMK)⁺ CD8 T cells as the dominant kidney-associated T cells which shared inflammation- and stress-related gene pathways with ABCs. Intrarenal GZMK⁺ CD8 T cells highly expressed <i>IFNG</i> and displayed strong communication with ABCs via the type II interferon (IFN) pathway. Intrarenal GZMK⁺ CD8 T cells and ABCs were largely co-localised within the tertiary lymphoid structure, and GZMK⁺ CD8 T cells potentially contributed to the differentiation of ABCs via IFN-γ and interleukin-21. <h3>Conclusions</h3> Our study revealed a potent extrafollicular B cell response linked with overactivation of GZMK⁺ CD8 T cells in the kidney of patients with LN, which may lead to innovative treatments for LN.