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[Preprint]. 2025 Dec 13:2025.12.10.693457. [Version 1] doi: 10.64898/2025.12.10.693457

The altered T cell landscape in Systemic Sclerosis patients is characterized by dysfunctional type 1 immunity

Victoria Volfson-Sedletsky, Hannah A DeBerg, Mitch L Fahning, Ian R Rifkin, Andreea M Bujor, Daniel J Campbell, Peter A Morawski, Anna C Belkina, Hans Dooms
PMCID: PMC12713596  PMID: 41427277

Abstract

Background

T cells in patients with chronic autoimmunity show features of dysfunction and exhaustion. However, the functionally-defined T cell subsets affected by these changes remain poorly characterized. Here, we sought to reveal aberrations in the composition, phenotype and function of canonical T cell subsets in the blood of Systemic Sclerosis (SSc) patients, compared to healthy subjects and Systemic Lupus Erythematosus (SLE) patients.

Methods

We developed a novel multidimensional flow-cytometry panel to simultaneously detect lineage-defining transcription factors, co-inhibitory receptors and other functional markers to characterize T cell subsets without in vitro restimulation. We compared T cell landscapes in SSc and SLE patients, and healthy subjects, using Optimized t-SNE and PhenoGraph algorithms. Cytokine production in patient samples was determined using intracellular cytokine staining. Transcriptomic analysis was performed to assess the role of IFN-γ in fibroblasts.

Results

The data show altered distribution and aberrant functional states of transcription factor-defined T cell subsets in patients with autoimmunity, with shared and unique features between diseases and their subtypes. Strikingly, SSc and SLE patients showed a severe deficiency in subsets of CD8 + and CD4 - CD8 - T cells that expressed T-bet, which is critical for IFN-γ-dependent type 1 immunity, and exhibited features of exhaustion. Moreover, TIGIT + Foxp3 + regulatory T cells, known to suppress Th1 responses, were selectively increased in SSc. Functionally, T cells from SSc patients produced less IFN-γ, a cytokine that suppresses pro-fibrotic gene expression in fibroblasts.

Conclusion

Our study demonstrates that SSc patients have a defective IFN-γ-producing T cell compartment that may lead to reduced anti-fibrotic T cell activity, enabling chronic fibrosis development.

Funding

National Institute of Allergy and Infectious Diseases/NIH, National Institute of Arthritis and Musculoskeletal Diseases/NIH, National Heart, Lung and Blood Institute/NIH, National Scleroderma Foundation, National Jewish Health

Full Text Availability

The license terms selected by the author(s) for this preprint version do not permit archiving in PMC. The full text is available from the preprint server.


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