Figure 4
Immunogenic effect of ciPTEC in direct co-culture with PBMC. (a) Representative dot plots and histograms of lymphocyte proliferation assays that were performed using carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled PBMC either untreated or activated with human T-Activator CD3/CD28 dynabeads (1 bead per cell). (b) Representative dot plots of CFSE-labeled PBMC proliferation after 5 days of co-culture with ciPTEC-U (untreated or pre-stimulated in various ways) in ratios 1:5, 1:15 or 1:30. Comparable dot plots were observed for the co-culture of PBMC and ciPTEC-T1. (c) PBMC proliferation after 5 days of co-culture in various ratios with untreated or pre-stimulated ciPTEC-U and –T1 cells, expressed as % of dividing cells. There were no differences in proliferation of PBMC from co-culture compared to untreated PBMC. Four independent co-culture experiments for ciPTEC-U and three for ciPTEC-T1 were performed. *p < 0.001 (One-way ANOVA, Tukey’s multiple comparison test) for PBMC proliferation induced by ConA (5 µg/ml), PHA-P (5 µg/ml) or T-Activator CD3/CD28 dynabeads (1 bead per cell) compared to untreated PBMC.
