Fig. 4

LPAs and autotaxin in ICD10 subgroups of MS patients and EAE mice. a Box plots showing the summed concentrations of all types of LPAs in serum of MS patients and healthy controls. The box represents the interquartile range, the whiskers show minimum to maximum, the line is the median. Patients were allocated to the groups according to available demographic data. “1^st attack” refers to patients with the first clinically manifest course of the disease that led to the diagnosis MS. “RRMS without relapse” are patients that were ‘symptom-free’ at the time of sampling after two or more previous relapses with or without prophylactic treatment. “RRMS with relapse” are patients with acute relapse with or without prophylactic treatment and “SPMS or PPMS” are patients with secondary or primary progressive disease. Data were compared with two-way ANOVA (factors “LPA” and “CD10-group”, posthoc adjustment of alpha according to Šidák. *P < 0.05, **P < 0.01, ***P < 0.001). b Scatter plots of autotaxin versus summed LPA concentrations in serum of MS patients and healthy controls and linear regression analyses showing significant associations of LPAs and autotaxin in both groups. c In analogy to (a), box plots show autotaxin concentrations in ICD10-subgroups. d Serum and spleen autotaxin concentrations in EAE and control mice, and in EAE mice treated with natalizumab 35 days after immunization. EAE was induced in female SJL/J mice with a standard PLP/PTX immunization protocol and serum samples were taken 35 days after immunization. Control mice got a CFA injection without PLP. Data were compared with one-way ANOVA. P < 0.05. e Autotaxin activity in primary splenocytes of EAE (SJL/J 35d after immunization) and control mice as assayed by conversion of C17-lysophosphatidylcholine (LPC) to the unnatural C17 LPA, which was analyzed by LC-MS/MS in cell culture supernatants. Cells were stimulated with PMA/Ionomycin (50/500 ng/ml) or vehicle in serum free medium for 2 h before adding the substrate, C17-LPC (100 μM). 50 μl samples of the supernatant were taken 0.5, 1 and 2 h after adding C17-LPC. Data were compared with 3-way ANOVA with the within subject factors “time” and between subject factors “stimulation” and “treatment”. Effects were significant for “time” P < 0.0001, “stimulation” P < 0.0001, “treatment” P < 0.001 and the respective interactions. Subsequently, corresponding groups were compared with 2-sided, unpaired t-tests employing a Bonferroni correction of alpha. The asterisk indicates a significant treatment effect, P = 0.0225. The data (mean ± SD) are results of 6 cultures per group. f LPA plasma concentration in naïve mice treated with vehicle or with the autotaxin inhibitor PF8380, which was injected i.p. 30 min before plasma sampling. Data were compared with two-way ANOVA (factors “LPA” and “treatment”) and subsequent 2-sided, unpaired t-tests for each LPA individually; *P < 0.05, *** P < 0.001