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• W4379801741 abstract "Background Cell surface glycosylation serves as a binding platform for endogenous lectin families such as galectins (Gals) or sialic acid immunoglobulin-like lectins (Siglecs) with well-known immunoregulatory functions. Several studies, including our own work [1] , have shown that Gals play an important role in pathogenesis of systemic lupus erythematosus (SLE) in animal models but their contribution to human SLE must be better elucidated. For these lectins to act, the appropriate glycan structures should be available on the target cells and should not be masked by alpha-2,6-sialylation which prevents binding of Gal-1, but not Gal-3. Although cancerous diseases are well-characterized by an altered glycome, limited information regarding cell surface glycosylation is gathered for autoimmune diseases like SLE. Objectives In the current study, we aim to analyse the ability of certain immune subsets to bind Gal-1, Gal-3, sialic acid binding C-type lectin (Siglec-1), the fucose binding Aleuria aurantia lectin (AAL), the sialic acid binding Sambucus nigra Agglutinin (SNA) lectin in SLE patients versus healthy controls. Methods We collected peripheral blood mononuclear cells (PBMCs) from 19 new or relapsing SLE patients with active disease (mean age: 47.5, SLEDAI-2K:15.8, anti-dsDNA level: 76.4 IU/ml) with no or minimal immunosuppressive therapy and 19 healthy age-matched controls. Multicolor antibody panel was designed to identify the main peripheral immune subsets: CD4+T-cells, CD8+ T-cells, CD4-/CD8- T-cells, CD4+/CD8+ T-cells; CD4+NKT-cells, CD8+NKT-cells, CD4-/CD8-NKT-cells, CD4+/CD8+NKT-cells; CD56+ NK cells; naive B-cells, memory B-cells, plasmablasts; classical monocytes, non-classical monocytes and intermediate monocytes. The five lectins (Gal-1, Gal-3, Siglec-1, AAL, SNA) were conjugated with different fluorophores for FACS. The binding of fluorochrome labelled lectins to each of these populations was assessed simultaneously by flow cytometry directly after thawing (resting state) and following activation. In case of T-cells, activation was achieved by a polyclonal activator (CytoStim), while B-cells and monocytes were activated by LPS. Results In our experiments with control PBMCs, titration and compensation of both antibodies and lectins were carried out and gating strategies were built to identify the above-mentioned immune subsets and their lectin binding characteristics. There were remarkable differences in lectin binding among different immune populations, e.g. classical and intermediate monocytes outperformed non-classical monocytes and lymphocytes in all lectin binding. Interestingly, naive B-cells were more sialylated (Median Fluorescence Intensity(MFI) SNA :2737) and bound less Gal-1 (MFI Gal-1 :5730) than both memory B-cells (MFI SNA :2368; MFI Gal-1 :16189) or plasmablasts (MFI SNA :1263; MFI Gal-1 :16515). Similarly, highest SNA (MFI SNA :144) and lowest Gal-1 (MFI Gal-1 :3658) binding was observed for CD4+ T-cells among the analysed T-cell subsets (MFI SNA range:12-120; MFI Gal-1 range: 4201-5460) or NK cells (MFI SNA not detectable; MFI Gal-1 :14288) or NKT-cells (MFI SNA range: not detectable-108; MFI Gal-1 range:3580-6327). Conclusion The focus of this study is to identify relevant changes in the cell surface glycosylation of SLE immune subsets which is a unique approach and could contribute to a better understanding of the glycobiology of lupus. Reference [1]Deak M,Hornung A et al. Immunobiology . 2014;220(4):483-489 Acknowledgements: NIL. Disclosure of Interests None Declared." @default.
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• W4379801741 date "2023-05-30" @default.
• W4379801741 modified "2023-10-18" @default.
• W4379801741 title "POS1419 SINGLE CELL ANALYSIS REVEALING THE LECTIN BINDING PATTERN OF T-CELLS, NKT-CELLS, NK CELLS, B-CELLS AND MONOCYTE SUBSETS IN SYSTEMIC LUPUS ERYTHEMATOSUS" @default.
• W4379801741 doi "https://doi.org/10.1136/annrheumdis-2023-eular.3549" @default.
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