Single-cell transcriptomic profiling reveals aberrant CD4⁺ naive T cell differentiation driving immune activation in Behçet’s uveitis - Abstract

Sep 30, 2026

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Behçet’s uveitis (BU) is a severe inflammatory eye manifestation of Behçet’s disease, marked by dysregulated T cell responses. The mechanisms behind T cell dysfunction and local immune activation in BU remain unclear. Single-cell RNA sequencing of peripheral blood mononuclear cells from BU patients and healthy controls was integrated with aqueous humor single-cell transcriptomes, then validated by qPCR, flow cytometry, and functional assays.

LanguageEnglish

CategoryHealthcare

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Reading Time40 min

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Abstract

Background

Methods

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Zhang et al. Journal of Translational Medicine (2026) 24:32 [https://doi.org/10.1186/s12967-025-07520-0](https://doi.org/10.1186/s12967-025-07520-0) Journal of Translational Medicine RESEARCH Open Access Single-cell transcriptomic profiling reveals aberrant CD4 ⁺ naiveT cell differentiation driving immune activation in Behçet’s uveitis Lu Zhang 1†, Yimeng Sun 1†, XinyuYao 1, Chun Yan 1, Yunfei Liu 1, Dijie Qiao 1, Ziyu Du2, Cong Hu 1, Fuying Guo3, Ziwei Meng 1 and Wei Chi 1,3* Abstract Background Behçet’s uveitis (BU) is a severe inflammatory ocular manifestation of Behçet’s disease, characterized by dysregulated T cell responses. However, the precise mechanisms underlying T cell dysfunction and local immune activation in BU remain poorly understood. Methods We performed single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) from BU patients and healthy controls (HCs) . These data were integrated with publicly available single-cell transcriptomes of aqueous humor (AH) samples from BU patients. Key findings were validated using qPCR, flow cytometry, and functional assays. Results We observed a significant depletion of naive CD4⁺ T cells in BU, accompanied by their differentiation toward terminal effector states characterized by IL-32 and CXCR4 upregulation. IL-32 was identified as a central regulator promoting Th1 and Th17 polarization. Pseudotime trajectory analysis revealed a progressive differentiation process from circulating CD4⁺ T cells toTh17-like inflammatory cells within the AH, enriched for NF-κB and JAK-STAT signaling pathways. CXCR4 expression was associated with T cell chemotaxis into the ocular microenvironment. Cell-cell interaction analyses further highlighted active crosstalk between macrophages and CD4⁺ T cells via the MIF-(CD74+CXCR4/CD44) signaling axis. Conclusion Our study reveals a coordinated mechanism linking systemic T cell dysregulation to localized ocular inflammation in BU. Targeting IL-32 and CXCR4 may represent promising strategies for restoring immune balance and preventing tissue damage in BU. Keywords Uveitis, Behçet’s disease, Single-cell RNA sequencing, T cells, Adaptive immunity †Lu Zhang andYimeng Sun contributed equally to this work as cofirst authors. *Correspondence: Wei Chi [chiwei@mail.sysu.edu.cn](chiwei@mail.sysu.edu.cn); [dr_chiwei@163.com](dr_chiwei@163.com) 1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangdong Provincial Clinical Research Center for Ocular Diseases, SunYat-Sen University, Guangzhou, Guangdong 510060, China 2Department of Ophthalmology, The First Hospital of Lanzhou University, Lanzhou, Gansu 730030, China 3Shenzhen Eye Center, Shenzhen Eye Hospital, Southern Medical University, Shenzhen, Guangdong 518043, China © The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creati](http://creati)[vecommons.org/l](vecommons.org/l)icenses/by-nc-nd/4.0/. Zhang et al. Journal of Tra

FAQs

What is the study’s main objective in Behçet’s uveitis?

To clarify the mechanisms driving T cell dysfunction and local immune activation in Behçet’s uveitis using single-cell transcriptomic profiling.

How were the experimental data generated and validated?

Single-cell RNA sequencing was performed on PBMCs from BU patients and healthy controls and integrated with publicly available aqueous humor single-cell data. Key findings were validated with qPCR, flow cytometry, and functional assays.

What differentiation change was observed in CD4⁺ T cells in BU?

Naive CD4⁺ T cells were depleted in BU and showed differentiation toward terminal effector states with IL-32 and CXCR4 upregulation, linked to Th1/Th17 polarization.

Which signaling pathways and immune interactions were implicated?

Pseudotime analysis indicated progressive differentiation toward Th17-like inflammatory cells enriched in NF-κB and JAK-STAT pathways. Cell-cell interaction analysis highlighted macrophage–CD4⁺ crosstalk via the MIF-(CD74+CXCR4/CD44) axis.

Sep 30, 2026

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