Regulation of Experimental Autoimmune Uveoretinitis by Anti-Delta-Like Ligand 4 Monoclonal Antibody

PURPOSE. To investigate the involvement of delta-like ligand (Dll) 4 in the development of experimental autoimmune uveoretinitis (EAU) in B10. RIII mice. METHODS. B10. RIII mice were immunized with interphotoreceptor retinoid binding protein (IRBP) peptide 161-180 in complete Freund's adjuvant together with intraperitoneal injection of Bordetella pertussis toxin. mRNA expressions of Notch receptors and their ligands in the eye were evaluated. To investigate the involvement of Dll in EAU, anti-Dll1, anti-Dll4, or control antibody (Ab) was intraperitoneally injected during both the induction and the effector phases or only the effector phase. Alternatively, mice were intraperitoneally injected with gamma-secretase inhibitor (GSI) or the control vehicle during the induction phase. Fourteen days after immunization, the eyes and spleens were harvested. The eyes were used for histologic and/or cytokine mRNA expression analysis, whereas the spleens were used for flow cytometric analysis, and antigen-recall proliferation and cytokine assays. RESULTS. Expression of Notch1, 2, 4, and Dll4 in the eye were upregulated by EAU induction. Anti-Dll4 Ab treatment during both the induction and effector phases, but not only the effector phase, significantly reduced the severity of EAU. IFN-gamma, IL-12p35, IL-17A, and TGF-beta mRNA expression in the eye were significantly attenuated by treatment with anti-Dll4 Ab. Splenocytes from anti-Dll4 Ab-treated mice showed significantly less proliferation and IL-17 production on antigen stimulation. Also, the severity of EAU was significantly reduced by gamma-secretase inhibitor treatment during the induction phase. CONCLUSIONS. Dll4-mediated Notch signaling during the sensitization is critical for the development of EAU. This can be a novel prophylactic target for autoimmune uveitis. (Invest Ophthalmol Vis Sci. 2011; 52: 8224-8230) DOI:10.1167/iovs.11-7756