Effect of selective IL-6 inhibitor Am-80 on experimental autoimmune encephalomyelitis in DA rats.
OBJECTIVE
To observe the role of interleukin (IL)-6 in the development of experimental autoimmune encephalomyelitis (EAE).


METHODS
DA rats were immunized by injecting bovine myelin basic protein (MBP). mRNA of cytokines, such as IL-6, IL-10, TNF-alpha, TGF-beta 1, IFN-gamma, and iNOS, were detected by RT-PCR. MBP was injected into ear to induce delayed type cutaneous hypersensitivity response (DTH). Histological studies were performed on the spinal cord with HE staining. Nitric oxide (NO) production from cultured murine macrophage clones was stimulated with LPS plus IFN-gamma.


RESULTS
DA rats developed EAE disease with a peak of severity on d 13 and d 14. Am-80 (1.0, 3.0 mg/kg), a selective IL-6 inhibitor, inhibited the symptoms in terms of deterioration as observed by the clinical score, body weight and histological findings, in a dose-related manner. A high dose of Am-80 (3.0 mg/kg for 12 d) did not completely inhibit the disease, but delayed the symptoms and enhanced the delayed response. By prolonging the duration of treatment (18 d), Am-80 inhibited the onset of EAE during administration, but the symptoms of EAE appeared after the administration was stopped. Am-80 administerd for 12 d inhibited the DTH response on d 11 but not on d 22. RT-PCR studies demonstrated a strong expression of IFN-gamma, IL-6, IL-10, TGF-beta 1, TNF-alpha, and iNOS mRNA in spinal cord 13 d after immunization. However IFN-gamma, IL-10, TNF-alpha, and iNOS mRNA expression (on d 13) was suppressed by Am-80, except in the case of IL-6, hence the effect of Am-80 on the expression of IL-6 mRNA was examined in additional experiments. After Am-80 was administered for 12 d or 18 d, the expression of IL-6 mRNA was inhibited on d 12 or d 18, but increased on d 13 or d 19, respectively.


CONCLUSIONS
These findings suggest that inhibition of EAE by Am-80 is initiated by inhibition of IL-6 production.
Wang
T
T
Department of Pharmacology, Gifu Pharmaceutical University, Gifu 502, Japan.
Nagai
H
H
Bouda
K
K
Matsuura
S
S
Takaoka
Y
Y
Niwa
S
S
Homma
T
T
Tanaka
H
H
Shudo
K
K
D000818:Animals; D001565:Benzoates / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000139:chemically induced / Q000378:metabolism*; D005260:Female; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D016753:Interleukin-10 / Q000096:biosynthesis / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D004676:Myelin Basic Protein; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D013116:Spinal Cord / Q000378:metabolism*; D013764:Tetrahydronaphthalenes / Q000494:pharmacology*; D000093242:Interleukin-6 Inhibitors
D001565:Benzoates; D016207:Cytokines; D007371:Interferon-gamma; D016753:Interleukin-10; D015850:Interleukin-6; D004676:Myelin Basic Protein; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger; D013764:Tetrahydronaphthalenes; D000093242:Interleukin-6 Inhibitors; C061133:tamibarotene; C496318:Nos2 protein, rat