FoxP3 expression by retinal pigment epithelial cells: transcription factor with potential relevance for the pathology of age-related macular degeneration
Ca-channels
610
Retinal Pigment Epithelium
Mice
Macular Degeneration
03 medical and health sciences
Research ; IL-1β ; Ca-channels ; CRISPR/Cas9 ; Phosphorylation ; RPE ; Age-related macular degeneration ; FoxP3 ; Immune barrier ; Immune privilege of the retina ; Medical and Health Sciences
FoxP3
IL-1 beta
Animals
Humans
Phosphorylation
Cas9
RC346-429
CRISPR/Cas9
Cells, Cultured
Immune barrier
Inflammation
0303 health sciences
Age-related macular degeneration
Research
Epithelial Cells
Forkhead Transcription Factors
ddc:
Rats
Immune privilege of the retina
IL-1β
CRISPR
RPE
Neurology. Diseases of the nervous system
Retinal Pigments
Transcription Factors
DOI:
10.1186/s12974-022-02620-w
Publication Date:
2022-10-22T13:05:13Z
AUTHORS (14)
ABSTRACT
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.
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CITATIONS (10)
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