Antigen-specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis

IF 4.6 2区医学 Q2 IMMUNOLOGY

Clinical & Translational Immunology Pub Date : 2025-05-15 DOI:10.1002/cti2.70036

Kaiser Alam, Arun Raina, Bibhuprasad Das, Sandhya Bhanja, Sayantan Ghosh, John V Forrester, Soumyava Basu

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Abstract

Objectives

Peripheral blood is frequently used to study the immune response in human uveitis because of the inaccessibility of ocular tissue samples. To determine whether peripheral blood immune cells accurately reflect the intraocular immune response, we compared the T-cell profiles and antigen-specific cytokine responses between paired vitreous and peripheral blood samples from patients with sight-threatening posterior uveitis.

Methods

We collected paired vitreous and peripheral blood mononuclear cells (PBMCs) from 24 patients with posterior uveitis. Multi-parametric flow cytometry was employed to identify surface and intracellular cytokine markers after activation with candidate antigenic peptides [Mycobacterium tuberculosis (MTb) peptides and retinal autoantigens]. Data were analysed through manual gating, unsupervised clustering and dimensionality reduction (FlowSOM, FlowJo).

Results

The CD8⁺/CD4⁺ ratio in a representative set of seven paired samples was higher in the vitreous than in PBMCs. Vitreous CD4⁺ and CD8⁺ cells displayed greater polyfunctional potential (TNFα⁺IFNγ⁺IL-2⁺ and PMA/ionomycin activation) than PBMCs. Upon antigen-specific activation in vitro, vitreous CD8⁺ T cells (but not CD4⁺ T cells) showed a stronger polyfunctional response than PBMCs against both MTb (in TB-immunoreactive patients) and retinal autoantigens. Unsupervised clustering identified 15 distinct CD3⁺ T-cell metaclusters, each with unique profiles in the vitreous and PBMCs. Significant cluster enrichment was observed among the vitreous infiltrating cells in TB-immunoreactive cases compared to non-TB uveitis, but no such enrichment was found among PBMCs in either patient cohort.

Conclusion

The vitreous T-cell compartment in this group of uveitis patients was functionally dominated by antigen-responsive cytotoxic CD8⁺ T cells and was distinct from the corresponding peripheral blood compartment.

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抗原特异性多功能细胞毒性T细胞在后葡萄膜炎中区分眼内免疫反应和外周血免疫反应

目的由于眼部组织样本难以获得，外周血常被用于研究人葡萄膜炎的免疫反应。为了确定外周血免疫细胞是否准确反映眼内免疫反应，我们比较了来自视力威胁后葡萄膜炎患者的配对玻璃体和外周血样本之间的t细胞谱和抗原特异性细胞因子反应。方法采集24例后葡萄膜炎患者的配对玻璃体和外周血单个核细胞（pbmc）。采用多参数流式细胞术鉴定候选抗原肽[结核分枝杆菌肽和视网膜自身抗原]激活后的细胞表面和细胞内细胞因子标志物。数据通过手动门控、无监督聚类和降维（FlowSOM、FlowJo）进行分析。结果玻璃体中CD8+/CD4+比值高于PBMCs。玻璃体CD4+和CD8+细胞表现出比pbmc更大的多功能潜能（TNFα+IFNγ+IL-2+和PMA/离子霉素激活）。在体外抗原特异性激活后，玻璃体CD8+ T细胞（而不是CD4+ T细胞）对MTb（在tb免疫反应性患者中）和视网膜自身抗原表现出比PBMCs更强的多功能反应。无监督聚类鉴定出15个不同的CD3+ t细胞元簇，每个元簇在玻璃体和pbmc中都有独特的特征。与非结核性葡萄膜炎患者相比，在结核免疫反应性患者的玻璃体浸润细胞中观察到显著的簇状富集，但在两组患者的pbmc中均未发现这种富集。结论本组葡萄膜炎患者玻璃体T细胞区室功能上以抗原反应性细胞毒性CD8+ T细胞为主，与相应的外周血区室不同。

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来源期刊

Clinical & Translational Immunology Medicine-Immunology and Allergy

CiteScore

12.00

自引率

1.70%

发文量

审稿时长

13 weeks

期刊介绍： Clinical & Translational Immunology is an open access, fully peer-reviewed journal devoted to publishing cutting-edge advances in biomedical research for scientists and physicians. The Journal covers fields including cancer biology, cardiovascular research, gene therapy, immunology, vaccine development and disease pathogenesis and therapy at the earliest phases of investigation.