FIGURE 8.

JW47 limits the accumulation of neurodegeneration and disability during relapsing autoimmune encephalomyelitis. ABH mice were injected with SCH in Freund's complete adjuvant on days 0 and 7 to induce paralytic EAE. A relapse was induced on day 28 during the first remission, using a further injection of SCH in Freund's incomplete adjuvant. Animals were treated daily from day 33 onward with either 1 mg/kg intraperitoneal JW47 (n = 14) in ethanol/cremophor/PBS (1:1:18) or vehicle (n = 12). The results represent the mean daily neurological score ± S.E. (error bars) (A) and the mean rotarod activity representing the mean ± S.E. time before falling/failing to stay on an accelerating rotarod before (on day 27) or after (on day 45) treatment with either vehicle or JW47 (B). *** p < 0.001 compared with vehicle treatment. C, axonal content in the spinal cord following treatment of relapsing EAE with JW47 1 mg/kg measured as neurofilament level adjusted for total protein content. EAE was induced with spinal cord homogenate in complete Freund's adjuvant on days 0 and 7, and a relapse was induced by reimmunization with spinal cord homogenate in complete Freund's adjuvant at day 28. Animals were randomized according to RotaRod performance score at day 27 to receive either vehicle (cremophor (Sigma)/alcohol/phosphate-buffered saline, 1:1:18) or 1 mg/kg intraperitoneal JW47 from day 33 postinfection just prior to the development of relapse at day 35 until day 47. Animals were killed, and the spinal cords were removed using hydrostatic pressure, and axonal content was measured using a quantitative neurofilament-specific ELISA. n = 11 untreated, n = 13 JW47-treated. Shown is the ratio of dephosphorylated (SMI32-reactive) neurofilament to hyperphosphorylated (SMI35-reactive) neurofilament as measured by ELISA in spinal cord homogenates from postrelapse untreated animals (n = 11) or JW47 (1 mg/kg)-treated animals (n = 13); ***, p < 0.001 adjusted for total protein level.