Abstract
Purpose :
To investigate the potential role of CUB domain-containing protein 1 (CDCP1) in the pathogenesis of experimental autoimmune uveitis (EAU).
Methods :
Multiple human primary RPE and RPE cell lines were examined for the presence of CDCP1 on the cell surface by flow cytometric analyses. CDCP1 knockdown (KD) RPEs were generated by transfecting the cells with lentivirus encoding CDCP1-specific shRNAs followed by flow sorting. Cytokine production profiles of the WT and CDCP1 KD RPEs were analyzed by ELISA. Both WT and CDCP1 KD RPE monolayers were also used in a transwell-based T cell migration assay. For in vivo experiments, the same numbers of WT uveitogenic T cells were adoptively transferred into naïve WT and CDCP1 KO mice, and the development of EAU was evaluated by various established ocular imaging techniques.
Results :
We found that CDCP1 is present on all RPE cells examined and its expression is upregulated after IFNg stimulation. The CDCP1-specific shRNA knocked down the expression of CDCP1 on the transfected RPEs, which resulted in reduced IL-6 production from the cells. In the in vitro T cell migration assays, fewer T cells migrated through the RPE monolayers when their CDCP1 expression was knocked down. More importantly, CDCP1 KO mice developed significantly less severe EAU with markedly reduced numbers of infiltrating leukocytes in the retina as assessed by fundoscopy, optical coherence tomography and ocular histopathological analyses.
Conclusions :
CDCP1 is present on RPEs and that it is required for efficient uveitogenic T cell infiltration through the outer blood-retinal barrier for the development of experimental autoimmune uveitis. These new insights suggest that CDCP1 could be a novel target for blocking autoreactive T cell infiltration into the retina for managing autoimmune uveitis.
This is a 2020 ARVO Annual Meeting abstract.