Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis
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 an...
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Wiley
2025-01-01
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| Series: | Clinical & Translational Immunology |
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| Online Access: | https://doi.org/10.1002/cti2.70036 |
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| author | Kaiser Alam Arun Raina Bibhuprasad Das Sandhya Bhanja Sayantan Ghosh John V Forrester Soumyava Basu |
| author_facet | Kaiser Alam Arun Raina Bibhuprasad Das Sandhya Bhanja Sayantan Ghosh John V Forrester Soumyava Basu |
| author_sort | Kaiser Alam |
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| description | 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. |
| format | Article |
| id | doaj-art-4a69efda3a374f2e91baf404d40bfa39 |
| institution | Kabale University |
| issn | 2050-0068 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical & Translational Immunology |
| spelling | doaj-art-4a69efda3a374f2e91baf404d40bfa392025-08-20T03:51:08ZengWileyClinical & Translational Immunology2050-00682025-01-01145n/an/a10.1002/cti2.70036Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitisKaiser Alam0Arun Raina1Bibhuprasad Das2Sandhya Bhanja3Sayantan Ghosh4John V Forrester5Soumyava Basu6Ocular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaOcular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaOcular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaOcular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaOcular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaOcular Immunology Group, Section of Infection and Immunity, Institute of Medical Sciences University of Aberdeen Aberdeen UKOcular Immunology Laboratory, Prof Brien Holden Eye Research Centre LV Prasad Eye Institute Hyderabad IndiaAbstract 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.https://doi.org/10.1002/cti2.70036antigenCD8 T cellsflow cytometrypolyfunctionalunsupervised clusteringuveitis |
| spellingShingle | Kaiser Alam Arun Raina Bibhuprasad Das Sandhya Bhanja Sayantan Ghosh John V Forrester Soumyava Basu Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis Clinical & Translational Immunology antigen CD8 T cells flow cytometry polyfunctional unsupervised clustering uveitis |
| title | Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| title_full | Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| title_fullStr | Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| title_full_unstemmed | Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| title_short | Antigen‐specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| title_sort | antigen specific polyfunctional cytotoxic t cells differentiate intraocular from peripheral blood immune responses in posterior uveitis |
| topic | antigen CD8 T cells flow cytometry polyfunctional unsupervised clustering uveitis |
| url | https://doi.org/10.1002/cti2.70036 |
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