Figure 4
Endothelial CD146 is necessary for the transmigration of lymphocytes in an in vitro BBB model.
(a) Flow cytometry analysis of T lymphocyte (CD4+, CD8+, TH1 and TH17 cells) transmigration across BBBECs that were treated with or without TNFα (50 ng/ml) in the presence of the AA98 antibody or control mIgG (50 μg/ml). Each data point represents the mean of each experiment with each donor (n = 8 donors). (b) The AA98 antibody blocked the migration of CD146+ T cells across the BBBEC monolayer. (c, d) Flow cytometry analysis of the T lymphocytes that transmigrated across BBBECs transfected with CD146 siRNA with or without rescuing CD146 expression. (e, f) Flow cytometry analysis of the lymphocytes that transmigrated across BBBECs in the presence of either the control mIgG or the AA98 antibody (50 μg/ml) and/or anti-ICAM-1 (10 μg/ml). (g) Pre-treatment of lymphocytes (CD146− and CD146+) with the AA98 antibody did not affect their transmigration. *, p < 0.05; **, p < 0.01, ***, p < 0.001, n.s., not significant ((a), paired Student's t-test; (b–g), unpaired Student's t-test). The data represent eight (a) or three (b–g) independent experiments.
