Fig 1: Increased KC/GRO expression in P2X7-/- mice post-status epilepticus and during epilepsy. (A) Experimental design. Wt and P2X7-/- mice were subjected to intra-amygdala KA-induced status epilepticus. Hippocampi were collected 8 h and 24 h post-lorazepam treatment. Blood samples were taken 8 h, 24 h, and 14 days post-KA. (B) Graph showing Racine scores (maximum reached) per 5 min period over 40 min after injection of KA for wt and P2X7-/- mice (n = 5 (wt) and 4 (P2X7-/-). (C) Heat map showing changes in cytokine concentrations in wt and P2X7-/- mice in control conditions in plasma and hippocampus 8 h following status epilepticus as measured via cytokine array. Blue color represents lower cytokine concentrations and yellow color higher cytokine concentrations. (D) KC/GRO levels in plasma of wt and P2X7-/- mice under control conditions and 8 h post-status epilepticus (SE) (control: N = 12 per genotype; post-SE: N = 15 (wt) and 18 (P2rx7-/-)) and during epilepsy (14 days post-KA injection) (control: N = 12 (wt) and 13 (P2rx7-/-); epilepsy: N = 16 (wt) and 19 (P2X7-/-)). (E) IL-1ß levels in plasma of wt and P2X7-/- mice under control conditions and 8 h post-status epilepticus (control: N = 12 (wt) and 13 (P2X7-/-); post-SE: N = 14 (wt) and 15 (P2X7-/-)) and during epilepsy (control N = 9 (wt) and 11 (P2X7-/-); epilepsy: N = 11 (wt) and 12 (P2X7-/-)). (F) IL-6 levels in plasma of wt and P2X7-/- mice under control conditions and 8 h post-status epilepticus (control: N = 12 (wt) and 13 (P2X7-/-); post-SE: N = 12 (wt) and 13 (P2X7-/-)) and during epilepsy (control: N = 6 (wt) and 7 (P2rx7-/-); epilepsy: N = 6 per genotype). (G) IL-18 levels in plasma of wt and P2X7-/- mice under control conditions and 8 h post-status epilepticus (control: N = 11 (wt) and 9 (P2rx7-/-); post-SE: N = 12 per genotype) and during epilepsy (control: N = 4 per genotype; epilepsy: N = 4 per genotype) (D–H). Data are represented as percentage to wt control. ANOVA with post-hoc Fisher correction test. * p < 0.05, ** p < 0.01.
Fig 2: Increased P2X7R expression in blood cells post-status epilepticus in mice. (A) Mice expressing P2X7 C-terminally fused to EGFP are subjected to intra-amygdala KA-induced status epilepticus. Blood was collected 1 h post-treatment with the anticonvulsant lorazepam. (B) Graph showing the histogram of the GFP-A (area) in control conditions and 1 h post-SE. The percentage of GFP+ cells is significantly increased in animals treated with intra-amygdala KA 1 h post-lorazepam (SE) when compared with vehicle-injected control mice (Ctrl) (N = 9 per group) (Unpaired Student’s t-test, p = 0.0493). (C) Forward side scatter indicating the distribution of the GFP+ cell population (in green). Note, size and complexity suggests the GFP+ cell population being composed mainly of monocytes. * p < 0.05.
Fig 3: Increased P2X7R protein levels in plasma of patients with TLE. (A) Patients were recruited from two hospitals: Marburg Hospital (Germany) and Beaumont Hospital (Ireland), and plasma samples were analyzed via P2X7R-detecting ELISA. (B) Bar chart showing P2X7R protein levels in plasma from temporal lobe epilepsy (TLE) patients (N = 30) in baseline conditions and 1 h post-seizure (N = 15) are higher when compared with healthy controls (Ctrl) (N = 34) and patients with psychogenic non-epileptic seizures (PNES) (N = 11). In addition, P2X7R protein levels were also higher in patients suffering from an episode of status epilepticus (SE) (N = 6) when compared with PNES patients. ANOVA with post-hoc Fisher correction. Data are given as percentage to control. (C) ROC curve analysis shows P2X7R plasma levels have a moderate sensitivity (60%) and good specificity (74%) for discriminating between healthy controls and TLE patients with an AUC of 0.64 at a cut-off of 119.2%. (D) ROC curve analysis demonstrating a good sensitivity (90%) and specificity (63%) for discriminating between TLE patients and patients with PNES with an AUC of 0.77 at a cut-off of 96.58%. (E) Dot plot showing P2X7R plasma levels in the same patients at baseline and 1 h post-seizure. Out of 10 patients, 7 showed increased P2X7R in plasma and 3 lower P2X7R plasma levels 1 h post-seizures when compared with the corresponding baseline levels. (F,G) No correlation of P2X7R plasma levels according to sex (F) (Control: p = 0.33; TLE: p = 0.92) or age (G) (Control: r2 = 0.03 p = 0.33; TLE: r2 = 0.003, p = 0.72). * p < 0.05, ** p < 0.01.
Fig 4: Scatter plot graphs of the Spearman correlation analysis. Correlation analysis between P2X7 and TNF-a, IL-1ß; between vitamin A and IgG, IgA, IgM; between 25(OH)D and IgG, IgA, IgM; were performed. Each symbol represents the measurement of one case. The continuous line shows the least-square linear regression
Fig 5: Evaluation of the serum levels of P2X7, vitamin A, and 25(OH)D3 among CP, nsMPP, and sMPP. The serum levels of P2X7 (Figure 1A), vitamin A (Figure 1B), and 25(OH)D (Figure 1C) were measured and compared among CPs, nsMPPs, and sMPPs. (n = 50 (CP), 84 (nsMPP) and 52 (sMPP). *<0.05; **<0.01; ***<0.001
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