Figure 7

Anti-TNFR1 treatment reduces TNFα-induced T cell adhesion and endothelial cell adhesion molecule expression. (A) An adhesion assay of DiI-labelled T cells (red) was performed using a human brain endothelial cell line (hCMEC/D3) which was grown to confluency and pre-activated as indicated prior to the assay. (C) Quantification revealed that 24 hour pre-treatment with huTNFα significantly increased the number of adherent T cells, but not when co-treated with ATROSAB. (B) Similarly, 24 hour treatment of hCMEC/D3 cells with huTNFα resulted in a robust production of VCAM-1 (red), which was essentially blocked by ATROSAB co-treatment, as quantified in (D). Dapi counter-staining (blue) indicates the endothelial cell nuclei. Measurement of surface (E) VCAM-1 and (G) ICAM-1 expression on hCMEC/D3 cells was assessed by flow cytometry, with quantification given in (F,H), respectively, showing a significant reduction in TNFα-induced upregulation by co-incubation with ATROSAB. Scale bars = 100 µm. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. (C,D), n = 4 per treatment, representative experiment of 2; (F), n = (control) 7, (TNFα) 7, and (TNFα + ATROSAB) 10; (H), n = (control) 3, (TNFα) 3, (TNFα + ATROSAB) 4.