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LO-037 Altered peripheral non-classic monocyte and NKT counts and CD8+ TCR diversity of systemic lupus erythematosus identified via integrated high-dimensional flow cytometry, CyTOF, metabolic profiling and TCR clonality analyses
  1. Anselm Mak1,
  2. Kok Siong Ang2 and
  3. Jinmiao Chen2
  1. 1Rheumatology, Medicine, National University of Singapore, Singapore
  2. 2Singapore Immunology Network (SIgN), AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH (A*STAR), Singapore


Background The phenotypic, signaling and metabolic diversities of leucocytes of systemic lupus erythematosus (SLE) impede comprehensive identification of immunopathologically-relevant alterations in leucocytes associated with active SLE. We aimed to identify these alteration signatures in leucocytes from patients with active SLE by an integrated platform comprising high-dimensional flow cytometry, cytometry by time of flight (CyTOF) and RNA sequencing (RNA-seq).

Methods Peripheral blood mononuclear cells (PBMCs) of SLE patients and healthy subjects were subjected to high-dimensional flow cytometry and CyTOF for studying alterations of myeloid cells and lymphocytes. Bulk RNA-seq was conducted for 8 sorted cell populations. Data were subjected to integrative analyses with Cytozoom that identified cellular signatures of active SLE. Differences in T-cell and B-cell receptor clonalities, cellular signaling and metabolic reaction pathways were studied based on SLE activity.

Results SLE patients with active disease had significantly lower CD14+CD16highCD86+HLA-DR+ and higher circulating CD3+CCR4+CXC3CR1+CD45RO+CD27+cells in their PBMCs (See figures 1A to 1Y), and higher TCR diversity in inactive CD8+ T cells compared to those with inactive disease, coupled with downregulated Sirtuin and upregulated oxidative phosphorylation and EIF2 signaling pathways in CD8+ T cells and B cells. Upregulated keratin sulphate synthesis reaction was observed in activated B cells, monocytes and plasmacytoid dendritic cells while the reaction was downregulated in inactive CD8+ T cells in active SLE patients

Conclusions An integrated analytical platform comprising high-dimensional cytometric analyses and RNA-seq demonstrated signatures that highlight the concerted and complex pathogenic signatures of non-classical monocytes, tissue-homing memory T cells and altered signaling/metabolic pathways of lymphocytes and myeloid cells in patients with active SLE.

Abstract LO-037 Figure 1

The paradigms in searching for flare-related cell types in Myeloid (A-L) and T/NK (M-Y) lineages, presented with UMAP and PAGA plots, and Leiden clustering

  • systemic lupus erythematosus
  • integrated platform
  • Biomarker

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