Altered frequency and function of mait cells in systemic sclerosis revealed by high dimensional mass cytometry and transcriptome analysis

Background: Systemic sclerosis (SSc) is an autoimmune disease characterised by excessive fibrosis of skin and internal organs, and vascular dysfunction. Association of T and B cell subsets have been reported in SSc, however there is lack of systematic studies of functional relations between immune cell subsets in this disease. This lack of mechanistic knowledge hampers targeted intervention. Objectives: In the current study we ought to determine differential immune cell composition and heterogeneity in peripheral blood of SSc patients and its impact on disease severity and progression. Methods: Mononuclear cells from blood of SSc patients with interstitial lung disease (ILD, n=10) or No ILD (n=10) and healthy controls (n=10) were analysed by mass cytometry using a 36 marker (cell-surface and intracellular) panel to aid in identification of major PBMC lineages including T cells, B cells, monocytes and NK cells and their subsets. Transcriptome analysis (m-RNA sequencing) was performed on sorted T and B cell subsets. Unsupervised clustering of mass cytometry data was performed using in-house developed analysis software MARVIS. This software combines dimension reduction and clustering steps to identify all possible cellular subsets. Further, custom R scripts helped in identifying nodes that were differentially expressed between the study groups and also phenotype of these nodes. Results: Unsupervised clustering performed revealed significant differences in the frequencies of T cell and B cell subsets. Most strikingly we identify a 3 fold decrease in frequencies of Va7.2+CD161+mucosal associated invariant T cells (MAIT) in SSc patients and 2 fold increase in total B cells, particularly CD19 +CD27 naive cells. A subset of memory CD8 +T cell, expressing CXCR3 was found to be increased in SSc patients as compared to healthy controls. Transcriptome analysis of sorted B cell and T cell subsets showed decrease in genes related to survival and increased expression of apoptotic genes in CD4,CD8 T and MAIT cells from SSc patients. Genes related to exhaustion and leukocyte migration were highly expressed in T cells from patients. Conclusions: This study provides an in depth analysis of systemic immune composition in SSc with the potential to delineate mechanisms of pathogenesis and identify diagnostic and/or therapeutic targets. This is the first demonstration of dysfunction of MAIT cells in SSc and further characterisation of their function in this context is required. References Denton CP, Khanna D. ‘Systemic sclerosis.’ Lancet 2017;390(10103):1685–1699. Fuschiotti P. Current perspectives on the role of CD8+ T cells in systemic sclerosis. Immunol Lett 2017. Liu M, et al. New insights into CD4(+) T cell abnormalities in systemic sclerosis.’ Cytokine Growth Factor Rev 2016;28:31–36. Sanges S, et al. ‘Role of B cells in the pathogenesis of systemic sclerosis.’ Rev Med Interne 2017;38(2):113–124. Disclosure of Interest: None declared DOI: 10.1136/annrheumdis-2018-eular.6082 Citation: Ann Rheum Dis, volume 77, supplement Suppl, year 2018, page A204Session: Triple T: T cells, technologies and therapies