Suppression of murine experimental autoimmune optic neuritis by mature dendritic cells transfected with calcitonin gene-related Peptide gene.
OBJECTIVE
Calcitonin gene-related peptide (CGRP) exhibits prominent anti-inflammatory actions. We examined whether CGRP-transfected dendritic cells (DC) prevent the development of experimental autoimmune optic neuritis (EAON) and experimental autoimmune encephalomyelitis (EAE).


METHODS
A human CGRP-expressing plasmid was constructed, and used to transfect C57BL/6 mouse bone marrow-derived matured DC (mDC) by electroporation


METHODS
Transfection efficiency was 50% with 80% cell viability. C57BL/6 mice were immunized with myelo-oligodendrocyte glycoprotein 35-55, and injected intravenously with CGRP-expressing mDC (CGRP gene-transfected group) or mock-transfected mDC (mock-transfected group) at the induction or effector phase. EAE was diagnosed clinically and EAON was assessed histopathologically. Delayed hypersensitivity was measured. Supernatants of spleen cell cultures were assayed for cytokines using ELISA. The CD4(+)CD25(+)Foxp3(+) fraction in spleen cells was analyzed using flow cytometry.


RESULTS
For gene therapy in the induction phase, EAE developed in 50% of mice in the CGRP-transfected group compared with 80% in the mock-transfected group, and the mean pathological score for EAON was 1 in the CGRP-transfected group compared with 2 in the mock-transfected group (P < 0.05). For gene therapy in the effector phase, the mean EAE clinical score (1.5 vs. 3.0) and mean EAON pathological score (1.0 vs. 2.0) were both lower in the CGRP-transfected group compared with the mock-transfected group (P < 0.05). Delayed hypersensitivity was suppressed significantly in the CGRP-transfected group. IL-10 production by spleen cells in the CGRP-transfected group increased independent of MOG concentration, compared with the mock-transfected group. Interestingly, the proportion of CD4(+)CD25(+)Foxp3(+) cells increased significantly (P < 0.05) in the CGRP-transfected group compared with the mock-transfected group.


CONCLUSIONS
Gene therapy with CGRP-expressing mDC was effective in suppressing the development of EAON and EAE.
Department of Ophthalmology, Tokyo Medical University, Shinjuku-ku, Tokyo, Japan.
Ryusaku
R
Matsuda
Takeshi
T
Kezuka
Chiharu
C
Nishiyama
Yoshihiko
Y
Usui
Yoshimichi
Y
Matsunaga
Yoko
Y
Okunuki
Naoyuki
N
Yamakawa
Hideoki
H
Ogawa
Ko
K
Okumura
Hiroshi
H
Goto
D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics* / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000502:physiology*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000517:prevention & control; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006968:Hypersensitivity, Delayed / Q000276:immunology; D007111:Immunity, Cellular; D051379:Mice; D008810:Mice, Inbred C57BL; D009444:Neuritis, Autoimmune, Experimental / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D009900:Optic Nerve / Q000473:pathology; D009902:Optic Neuritis / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D016207:Cytokines; D015740:Calcitonin Gene-Related Peptide