Fig. 6: sCSF1Rinh significantly suppresses clinical disease progression in the NOD-EAE model.
From: CSF1R signaling is a regulator of pathogenesis in progressive MS
a Disease scores demonstrate the initial relapsing/remitting and progressive stages of the MOG-induced NOD-EAE model. Upon treatment with sCSF1Rinh at Day 28, mean disease scores were significantly reduced. Data points represent the mean and standard error of the mean (n = 12–13). A two-way ANOVA was used to determine the statistical significance of the differences between vehicle and sCSF1Rinh treated groups. For sCSF1Rinh at 25 mg/kg, p ≤ 0.05 on Day 36–37, 39–50, 52–61 while sCSF1Rinh at 100 mg/kg had p ≤ 0.05 on Day 35–61. b Area under the curve was calculated for the clinical disease course and sCSF1Rinh significantly reduces paralytic symptoms at both doses. Data points represent the area under the curve for each animal and graphical columns represent the mean and standard error. Statistical significance was determined by a one-way ANOVA and p values are indicated by *p < 0.05, ***p < 0.001. c Iba1 immunohistochemistry was used to visualize microglia and infiltrating macrophages in the spinal cord and sCSF1Rinh significantly reduced the Iba1+ area. Data points represent the Iba1+ area for each animal while graphical columns represent the mean and standard error. Statistical significance was determined by a one-way ANOVA and p values are indicated by ***p < 0.001, ****p < 0.0001. d Inflammatory cytokine production in the spinal cord was assessed with ELISA and sCSF1Rinh treatment at 25 mg/kg significantly reduced IL-6, IP-10, IL-1β, and MCP-1. Data points represent the protein quantification from a single animal. Graphical columns represent the mean and standard error (n = 3, 9, and 8, respectively). A one-way ANOVA was performed to determine statistical significance and p values are indicated by *p < 0.05. e Quantitative NanoString nCounter mRNA analysis of microglial gene expression in the spinal cord of the NOD-EAE model. sCSF1Rinh treatment at 25 mg/kg significantly increased the expression of homeostatic genes such as TMEM119, P2RY12, and CX3CR1 while also reducing the expression of pro-inflammatory genes such as SPP1, Lilrb4, and Fabp5. Data points represent the gene expression for each animal while graphical columns represent the mean and standard error. Statistical significance was determined by a one-way ANOVA and p values are indicated by *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. f Bielschowsky’s silver staining reveals a decrease in axons within the spinal cord of NOD-EAE mice and sCSF1Rinh significantly reduced the axonal loss pathology score demonstrating neuroprotection in this model. Statistical significance was determined by a one-way ANOVA and p values are indicated by **p < 0.01. Scale bar: 200 µm.
