Figure 4

Ectopic expression of CD20 on T cells associates with altered transmigration and adhesive properties but does not alter phenotype or response to TCR/antigen stimulation. T cells were transduced with the CD20-encoding retroviral vectors (mean transduction efficiency of 50%). (A) CD20- and CD20 + T cells were gated and the percentage CD3, CD25, CD69 and HLA-DR and (B) the mean fluorescence intensity (MFI) was determined. The bars represent the mean ± SD of three independent experiments. (C) The cow milk peptide (CMP) CD4 + T cell clone (BT55) was transduced with the CD20-encoding retroviral vector and subsequently purified. Different concentrations of CMP (100 μg–0.001 μg) were loaded on autologous and allogeneic EBV-B cells and presented to the transduced and the non-transduced cells. IL-13 production was measured upon stimulation of the BT55 clone with increasing concentrations of CMP. The mean ± SD of three independent experiments are shown. A 2way ANOVA was performed to determine significance. (D) For the migration assay HS-5 stromal cells were seeded in a 24-wells plate and after 24 h of incubation at 37 °C, T cells with or without CD20 were added onto a transwell system (5 μm pore size). These transwell systems were placed into 24-wells plate containing the stromal cells or as a control the medium. After 24 h the percentage of the migrated cells was determined by FACS analyses.