AAV8-Mediated Retinal PD-L1 Gene Transfer Attenuates Experimental Autoimmune Uveitis by Restoring Local Immune Tolerance.

PURPOSE: To investigate the therapeutic potential of the programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 (PD-L1) immune checkpoint pathway in experimental autoimmune uveitis (EAU) and to elucidate its role in regulating the retinal immune microenvironment.

METHODS

Seven days before EAU induction, Lewis rats received subretinal injections of adeno-associated virus serotype 8 (AAV8)/PD-L1. EAU was induced by immunization with interphotoreceptor retinoid-binding protein (IRBP) in complete Freund's adjuvant. Clinical inflammation scores were recorded on days 8, 10, 12, and 14 post-immunizations. Retinal function was evaluated by electroretinography (ERG) on day 12, and structural changes were assessed by optical coherence tomography (OCT). On day 14, eyes were harvested for histopathological examination, immunofluorescence, and flow cytometric analyses of leukocyte infiltration and cytokine expression. Normal rats injected with AAV8/PD-L1 were examined to assess retinal safety.

RESULTS

AAV8/PD-L1 markedly enhanced PD-L1 expression in the retina, resulting in significantly reduced intraocular inflammation and tissue damage compared with untreated EAU rats. The PD-L1 group showed lower anterior chamber inflammation scores (P < 0.05), decreased retinal T-cell and leukocyte infiltration (P < 0.05), and downregulated expression of IL-17. No structural or functional abnormalities were observed in normal eyes treated with AAV8/PD-L1.

CONCLUSIONS

AAV8-mediated PD-L1 overexpression suppresses EAU progression by restoring retinal immune tolerance, attenuating inflammation, and preserving visual function. This approach offers a promising gene therapy strategy for local immune modulation in ocular autoimmune diseases.