Transcriptional and Clonal Characterization of Cytotoxic T Cells in Crescentic Glomerulonephritis

Abstract

Significance Statement T-cell infiltration is a hallmark of crescentic GN (cGN), often caused by ANCA-associated vasculitis. Pathogenic T-cell subsets, their clonality, and downstream effector mechanisms leading to kidney injury remain to be fully elucidated. Single-cell RNA sequencing and T-cell receptor sequencing revealed activated, clonally expanded cytotoxic CD4+ and CD8+ T cells in kidneys from patients with ANCA-associated cGN. In experimental cGN, kidney-infiltrating CD8+ T cells expressed the cytotoxic molecule, granzyme B (GzmB), which induced apoptosis in renal tissue cells by activation of procaspase-3, and aggravated disease pathology. These findings describe a pathogenic function of (clonally expanded) cytotoxic T cells in cGN and identify GzmB as a mediator and potential therapeutic target in immune-mediated kidney disease. Background Crescentic GN (cGN) is an aggressive form of immune-mediated kidney disease that is an important cause of end stage renal failure. Antineutrophilic cytoplasmic antibody (ANCA)-associated vasculitis is a common cause. T cells infiltrate the kidney in cGN, but their precise role in autoimmunity is not known. Methods Combined single-cell RNA sequencing and single-cell T-cell receptor sequencing were conducted on CD3+ T cells isolated from renal biopsies and blood of patients with ANCA-associated cGN and from kidneys of mice with experimental cGN. Functional and histopathological analyses were performed with Cd8a −/− and GzmB −/− mice. Results Single-cell analyses identified activated, clonally expanded CD8+ and CD4+ T cells with a cytotoxic gene expression profile in the kidneys of patients with ANCA-associated cGN. Clonally expanded CD8+ T cells expressed the cytotoxic molecule, granzyme B (GzmB), in the mouse model of cGN. Deficiency of CD8+ T cells or GzmB ameliorated the course of cGN. CD8+ T cells promoted macrophage infiltration and GzmB activated procaspase-3 in renal tissue cells, thereby increasing kidney injury. Conclusions Clonally expanded cytotoxic T cells have a pathogenic function in immune-mediated kidney disease.