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Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: High levels of endothelial ICAM-1 prohibit natalizumab mediated abrogation of CD4+ T cell arrest on the inflamed BBB under flow in vitro

Fig. 5

High levels of ICAM-1 compared to VCAM-1 allow for T cell arrest in the presence of natalizumab. A Number of arrested human CD4+ Th1* cells on equimolar concentrations of immobilized recombinant VCAM-1 (1X, 1.54 μg/mL), ICAM-1 (1X, 1.14 μg/mL), and combined VCAM-1 (1X) / ICAM-1 (1X) under flow condition. B Number of adhered Th1* cells on immobilized recombinant VCAM-1 (1X), 10-times more ICAM-1 (10X, 11.4 μg/mL) and combined VCAM-1 (1X) / ICAM-1 (10x) under flow condition. A, B Th1* cells were treated with 1 μg/mL natalizumab (NTZ) and/or 1 μg/mL of an anti-β2-integrins antibody prior experiment. An isotype control antibody was used as internal control (1 μg/mL Ctrl). Mean crawling speed (C), distance (D) and Euclidian distance (E) of Th1* cells on equimolar concentration of immobilized recombinant VCAM-1 (1X), ICAM-1 (1X), and combined VCAM-1 (1X) / ICAM-1 (1X) under physiological flow conditions are depicted for one representative experiment in each group (> 100 cells per group were analysed). Each data point represents the velocity, the distance, and the Euclidian distance of one cell. E, F Directionality of Th1* cell crawling on immobilized recombinant VCAM-1, ICAM-1, and combined VCAM-1 / ICAM-1 under physiological flow condition. Results for equimolar concentrations of VCAM-1 (1X) and ICAM-1 (1X) (E), and for 10-times more ICAM-1 (10X) are shown (F). Directionality of Th1* cell crawling is expressed as xFMI (xFMI = Dx/Dacc, Dx: straight x-axis distance covered by the T cell, Dacc: accumulated total distance of T cell movement). Direction of the physiological flow was along the x-axis from plus to minus and is indicated by an arrow (yellow). A, B, E, F Each figure shows the mean ± SEM of 3 independent experiments. Statistical analysis: one-way ANOVA followed by Tukey’s multiple-comparison test. C, D Each figure shows the mean ± SEM of one representative experiment per group (> 100 cells per condition were analysed). Statistical analysis: one-way ANOVA followed by Dunn’s multiple-comparison test (Kruskal–Wallis test) (p < 0.05 = *, p < 0.01 = **, p < 0.001 = ***, p < 0.0001 = ****)

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