Suppression of Murine Experimental Autoimmune Uveoretinitis by Mature Dendritic Cells Cultured With Calcitonin Gene–Related Peptide

Abstract

Abstract: : Purpose:To investigate the role of the suppression in murine experimental autoimmune uveoretinitis (EAU) by mature dendritic cells (DC) cultured with calcitonin gene–related peptide (CGRP). Methods:Bone marrow cells derived from C57BL/6 mice were cultured with GM–CSF for six days, and differentiated CD11⁺ immature DC were purified by autoMACS. Immature DC were further cultured with LPS (1 µg/ml) overnight, and then with human interphotoreceptor retinoid binding protein (IRBP) residues 1–20 (hIRBP–p) in the presence of CGRP or TGF–ß2. EAU was induced by immunization with hIRBP–p in Freund’s adjuvant and pertussis toxin in mice. At the same time, hIRBP–p–immunized mice were injected the mature DC cultured with CGRP or TGF–ß2 intravenously. On day 19 after immunization, hIRBP–p–specific delayed hypersensitivity (DH) were measured. On day 21 after immunization, animals were sacrificed, and assessed the extent of EAU pathologically. Results:EAU and antigen–specific DH was predominantly suppressed by injection of mature DC cultured with CGRP or TGF–ß2 and hIRBP–p. Incidence of EAU on control group was 87% (13 of 15 EAU mice), TGF–ß2 culture group was 60% (9 of 15 EAU mice), and CGRP culture group was 28% (5 of 18 EAU mice). Conclusions:It was demonstrated that mature DC cultured with CGRP play an immunosuppressive role in development of EAU, which were more effective than those cultured with TGF–ß2.

Keywords: immunomodulation/immunoregulation • immune tolerance/privilege • uveitis-clinical/animal model