Background: Sjögren’s disease (SjD) is a systemic autoimmune disorder characterized by lymphocytic infiltrations in the exocrine glands leading to tissue damage and sicca symptoms. Although the pathology of the disease is not yet well elucidated, type IFN Is, B-cell hyperactivation, aberrant B-T-cell cross-talk, and altered composition of T-cell subpopulations are thought to be hallmarks of the pathogenesis of SjD. Previously, we reported that naïve CD4+ T-cells exhibit signs of immune cell aging and impaired homeostatic proliferation, thus leading to peripheral lymphopenia, a frequent finding in SjD patients [1].

Objectives: Since metabolic changes are associated with immune cell senescence and autoimmunity, we evaluated metabolic and immunological parameters in naive CD4+ T cells of SjD patients using a multi-omics approach and in vitro functional assays.

Methods: Peripheral naive CD4+ T-cells were isolated from venous blood of SjD patients fulfilling 2016 ACR/EULAR classification criteria and matched healthy control subjects (HC). Material from each donor was used for RNA sequencing and HPLC/HRMS untargeted metabolomics. Naive CD4+ T-cells were cultured in vitro for 14 days to assess their responsiveness to homeostatic T-cell cytokine IL7. Cells treated with IL7 in combination with type I interferons (IFN I) IFN α2a + IFN-β1a at a concentration of 500U – 1500 U/ml were used to assess the effect of IFN I on homeostatic proliferation. Immunological, metabolic and aging parameters were measured at the end of the experiment.

Results: Transcriptome analysis showed 100 differentially expressed genes (DEG) between the SjD and HC samples, with 85 genes being upregulated. Unsupervised clustering using DEG clearly distinguished diseased from healthy samples. Manual gene annotation, gene ontology analysis and pathway enrichment analysis identified majority of genes being type I interferon- induced genes related to IFN α/β signaling and cellular sensing of virus. A newly developed Immune Response Enrichment Analysis (IREA) predicted 15 cytokines as potential drivers of transcriptional changes in SjD cells. IFN α1 and IFN β showed the highest enrichment score. Gene Set Enrichment Analysis (GSEA) identified IFN α/β signaling and genes related to a presence of virus as the most positively enriched in the SjD phenotype. Metabolomic analysis identified 277 metabolites covering important biological classes, namely amino acids and peptides, carbohydrates, energy metabolism metabolites, fatty acids, lipids and steroids, nucleotides, vitamins and others. All metabolite classes detected were increased in the SjD samples except lipids. The decreased lipid content, especially in the phosphatidylcholine (PC) class was cofounded with increased LysoPC and fatty acids, two products of the PC hydrolysis. This change was confirmed in the GSEA analysis with positive enrichment score for fatty acid metabolic processes and long chain fatty acid biosynthetic processes. Following transcriptomic analysis, we evaluated the effect of IFN I on naïve CD4+ T-cells function. Confirming our previous data, SjD naïve cells did not respond to exogenous IL7 stimulation and showed profound decrease in homeostatic proliferation compared to HC cells. Co-treatment of IL7 with type I interferons in HC cells recapitulated SjD as reduced homeostatic proliferation and induction of cellular senescence measured as senescence-associated β-galactosidase activity. This was accompanied by mitochondrial hyperpolarization and reduced glucose uptake.

Conclusion: Multi-omics analysis of naïve CD4+ T-cells from Sjögren´s patients showed type IFN I signature and increased activity of multiple metabolic pathways. Treatment of healthy cells with IL7 + type IFN I reduced their homestatic proliferation and induced the cellular senescence described in SjD cells. We propose, type I interferons drive naïve CD4+ T-cells dysfunction and senescence via metabolic reprograming of mitochondrial and glucose pathways.

REFERENCES: [1] Fessler J, Fasching P, Raicht A, Hammerl S, Weber J, Lackner A, Hermann J, Dejaco C, Graninger WB, Schwinger W, Stradner MH. Lymphopenia in primary Sjögren’s syndrome is associated with premature aging of naïve CD4+ T cells. Rheumatology (Oxford). 2021 Feb 1;60(2):588-597. doi: 10.1093/rheumatology/keaa105. PMID: 32227243.

Acknowledgements: NIL.

Disclosure of Interests: None declared.

© The Authors 2025. This abstract is an open access article published in Annals of Rheumatic Diseases under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/ ). Neither EULAR nor the publisher make any representation as to the accuracy of the content. The authors are solely responsible for the content in their abstract including accuracy of the facts, statements, results, conclusion, citing resources etc.