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• W3204282415 abstract "Abstract Background: Neutrophil extracellular traps (NETs) from patients with Systemic Lupus Erythematosus (SLE) are characterized by lower ubiquitylation and myeloperoxidase (MPO) as a substrate. The structural and functional effect of such modification and if there are additional post-translational modifications (PTMs) are unknown. Methods: To assess the expression and functional role of PTMs in NETs of patients with SLE; reactivation, proliferation and cytokine production was evaluated by flow cytometry using co-cultures with dendritic cells (DC) and CD4 + from SLE patients and healthy controls. The impact of ubiquitylation on MPO was assessed by molecular dynamics. The expression of ISG15 in NETs was evaluated by immunofluorescence and Western Blot. Results: Fifteen patients with SLE and 10 healthy controls were included. In the co-cultures of CD4 + lymphocytes with DC stimulated with ubiquitylated MPO or recombinant MPO, a higher expression of IFNγ and IL17A was found in CD4 + from SLE patients (p < 0.05). Furthermore, with DC stimulated with ubiquitylated MPO a trend towards increased expression of CD25 and Ki67 was found in lupus CD4 + lymphocytes, while the opposite was documented in controls (p < 0.05). Through molecular dynamics we found the K129-K488-K505 residues of MPO as susceptible to ubiquitylation. Ubiquitylation affects the hydration status of the HEME group depending on the residue to which it is conjugated. R239 was found near by the HEME group when the ubiquitin was in K488-K505. In addition, we found greater expression of ISG15 in the SLE NETs vs controls (p < 0.05), colocalization with H2B (r = 0.78) only in SLE samples and increased production of IFNγ in PBMCs stimulated with lupus NETs compared to healthy controls NETs. Conclusion: The ubiquitylated MPO has a differential effect on the induction of reactivation and proliferation of CD4 + lymphocytes in patients with SLE, which may be related to structural changes by ubiquitylation at the catalytic site of MPO. Besides a lower ubiquitylation pattern, NETs of patients with SLE are characterized by the expression of H2B conjugated with ISG15, and the induction of IFNγ by Th1 cells." @default.
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• W3204282415 date "2020-07-24" @default.
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• W3204282415 title "The role of post-translational modifications by ubiquitin/ISG15 tags in exposed proteins during NETosis: Immunogenic or tolerogenic signaling process in Systemic Lupus Erythematosus?" @default.
• W3204282415 doi "https://doi.org/10.21203/rs.3.rs-44910/v1" @default.
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