FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration
Abstract Background Forkhead-Box-Protein P3 (FoxP3) is a transcription factor and marker of regulatory T cells, converting naive T cells into Tregs that can downregulate the effector function of other T cells. We previously detected the expression of FoxP3 in retinal pigment epithelial (RPE) cells,...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-022-02620-w |
| _version_ | 1811336297633021952 |
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| author | Ahmad Samir Alfaar Lucas Stürzbecher Maria Diedrichs-Möhring Marion Lam Christophe Roubeix Julia Ritter Kathrin Schumann Balasubramaniam Annamalai Inga-Marie Pompös Bärbel Rohrer Florian Sennlaub Nadine Reichhart Gerhild Wildner Olaf Strauß |
| author_facet | Ahmad Samir Alfaar Lucas Stürzbecher Maria Diedrichs-Möhring Marion Lam Christophe Roubeix Julia Ritter Kathrin Schumann Balasubramaniam Annamalai Inga-Marie Pompös Bärbel Rohrer Florian Sennlaub Nadine Reichhart Gerhild Wildner Olaf Strauß |
| author_sort | Ahmad Samir Alfaar |
| collection | DOAJ |
| description | Abstract Background Forkhead-Box-Protein P3 (FoxP3) is a transcription factor and marker of regulatory T cells, converting naive T cells into Tregs that can downregulate the effector function of other T cells. We previously detected the expression of FoxP3 in retinal pigment epithelial (RPE) cells, forming the outer blood–retina barrier of the immune privileged eye. Methods We investigated the expression, subcellular localization, and phosphorylation of FoxP3 in RPE cells in vivo and in vitro after treatment with various stressors including age, retinal laser burn, autoimmune inflammation, exposure to cigarette smoke, in addition of IL-1β and mechanical cell monolayer destruction. Eye tissue from humans, mouse models of retinal degeneration and rats, and ARPE-19, a human RPE cell line for in vitro experiments, underwent immunohistochemical, immunofluorescence staining, and PCR or immunoblot analysis to determine the intracellular localization and phosphorylation of FoxP3. Cytokine expression of stressed cultured RPE cells was investigated by multiplex bead analysis. Depletion of the FoxP3 gene was performed with CRISPR/Cas9 editing. Results RPE in vivo displayed increased nuclear FoxP3-expression with increases in age and inflammation, long-term exposure of mice to cigarette smoke, or after laser burn injury. The human RPE cell line ARPE-19 constitutively expressed nuclear FoxP3 under non-confluent culture conditions, representing a regulatory phenotype under chronic stress. Confluently grown cells expressed cytosolic FoxP3 that was translocated to the nucleus after treatment with IL-1β to imitate activated macrophages or after mechanical destruction of the monolayer. Moreover, with depletion of FoxP3, but not of a control gene, by CRISPR/Cas9 gene editing decreased stress resistance of RPE cells. Conclusion Our data suggest that FoxP3 is upregulated by age and under cellular stress and might be important for RPE function. |
| first_indexed | 2024-04-13T17:37:24Z |
| format | Article |
| id | doaj.art-daec2053b2844090a0f870d522137c96 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-04-13T17:37:24Z |
| publishDate | 2022-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-daec2053b2844090a0f870d522137c962022-12-22T02:37:19ZengBMCJournal of Neuroinflammation1742-20942022-10-0119112310.1186/s12974-022-02620-wFoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degenerationAhmad Samir Alfaar0Lucas Stürzbecher1Maria Diedrichs-Möhring2Marion Lam3Christophe Roubeix4Julia Ritter5Kathrin Schumann6Balasubramaniam Annamalai7Inga-Marie Pompös8Bärbel Rohrer9Florian Sennlaub10Nadine Reichhart11Gerhild Wildner12Olaf Strauß13Experimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität, Berlin Institute of Health, Humboldt-UniversityExperimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität, Berlin Institute of Health, Humboldt-UniversitySection of Immunobiology, Department of Ophthalmology, University Hospital, LMU MunichInstitut de La Vision, Sorbonne Université, INSERM, CNRSInstitut de La Vision, Sorbonne Université, INSERM, CNRSInstitut Für Med. Mikrobiologie, Immunologie Und Hygiene, TU MünchenInstitut Für Med. Mikrobiologie, Immunologie Und Hygiene, TU MünchenDepartment of Ophthalmology, College of Medicine, Medical University South CarolinaExperimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität, Berlin Institute of Health, Humboldt-UniversityDepartment of Ophthalmology, College of Medicine, Medical University South CarolinaInstitut de La Vision, Sorbonne Université, INSERM, CNRSExperimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität, Berlin Institute of Health, Humboldt-UniversitySection of Immunobiology, Department of Ophthalmology, University Hospital, LMU MunichExperimental Ophthalmology, Department of Ophthalmology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität, Berlin Institute of Health, Humboldt-UniversityAbstract Background Forkhead-Box-Protein P3 (FoxP3) is a transcription factor and marker of regulatory T cells, converting naive T cells into Tregs that can downregulate the effector function of other T cells. We previously detected the expression of FoxP3 in retinal pigment epithelial (RPE) cells, forming the outer blood–retina barrier of the immune privileged eye. Methods We investigated the expression, subcellular localization, and phosphorylation of FoxP3 in RPE cells in vivo and in vitro after treatment with various stressors including age, retinal laser burn, autoimmune inflammation, exposure to cigarette smoke, in addition of IL-1β and mechanical cell monolayer destruction. Eye tissue from humans, mouse models of retinal degeneration and rats, and ARPE-19, a human RPE cell line for in vitro experiments, underwent immunohistochemical, immunofluorescence staining, and PCR or immunoblot analysis to determine the intracellular localization and phosphorylation of FoxP3. Cytokine expression of stressed cultured RPE cells was investigated by multiplex bead analysis. Depletion of the FoxP3 gene was performed with CRISPR/Cas9 editing. Results RPE in vivo displayed increased nuclear FoxP3-expression with increases in age and inflammation, long-term exposure of mice to cigarette smoke, or after laser burn injury. The human RPE cell line ARPE-19 constitutively expressed nuclear FoxP3 under non-confluent culture conditions, representing a regulatory phenotype under chronic stress. Confluently grown cells expressed cytosolic FoxP3 that was translocated to the nucleus after treatment with IL-1β to imitate activated macrophages or after mechanical destruction of the monolayer. Moreover, with depletion of FoxP3, but not of a control gene, by CRISPR/Cas9 gene editing decreased stress resistance of RPE cells. Conclusion Our data suggest that FoxP3 is upregulated by age and under cellular stress and might be important for RPE function.https://doi.org/10.1186/s12974-022-02620-wIL-1βCa-channelsCRISPR/Cas9PhosphorylationRPEAge-related macular degeneration |
| spellingShingle | Ahmad Samir Alfaar Lucas Stürzbecher Maria Diedrichs-Möhring Marion Lam Christophe Roubeix Julia Ritter Kathrin Schumann Balasubramaniam Annamalai Inga-Marie Pompös Bärbel Rohrer Florian Sennlaub Nadine Reichhart Gerhild Wildner Olaf Strauß FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration Journal of Neuroinflammation IL-1β Ca-channels CRISPR/Cas9 Phosphorylation RPE Age-related macular degeneration |
| title | FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration |
| title_full | FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration |
| title_fullStr | FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration |
| title_full_unstemmed | FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration |
| title_short | FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration |
| title_sort | foxp3 expression by retinal pigment epithelial cells transcription factor with potential relevance for the pathology of age related macular degeneration |
| topic | IL-1β Ca-channels CRISPR/Cas9 Phosphorylation RPE Age-related macular degeneration |
| url | https://doi.org/10.1186/s12974-022-02620-w |
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