| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Research Abstract |
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). 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-B2. 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-B2 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. EAU and antigen-specific DH was predominantly suppressed by injection of mature DC cultured with CGRP or TGF-B2 and hIRBP-p. Incidence of EAU on control group was 87% (13 of 15 EAU mice), TGF-B2 culture group was 60% (9 of 15 EAU mice), and CGRP culture group was 28% (5 of 18 EAU mice). On the next experiments, we prepared CGRP gene-transfect-DC by electroporation methods. As the results, CGRP gene-transfect-DC can suppress EAU, and suppress DH spepific for IRBP-p. It was demonstrated that mature DC cultured with CGRP and CGRP gene-transfect-DC play an immunosuppressive role in development of EAU, which were more effective than those cultured with TGF-B2.
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