Figure 5

Block of allogeneic CD8 ⁺ T cell proliferation upon coculture with RCC cells pretreated with IL-4 and TNFα . A-C) Hal31RC cells were treated with either IL-4 or TNFα alone and with the combination of both cytokines. CD8⁺ T cells from allogeneic PBMCs were sorted, CFDA-SE labeled and cocultured with cytokine pretreated RCC31Hal. A) Representative FACS histogramm plots of proliferated CD8⁺ Tcells cocultured with untreated (u RCC) as compared to IL-4 or TNFα pretreated Hal31RC (4 + T RCC) are shown. The division index (DI) is given. B) Summary of CD8⁺ T cell proliferation on cytokine pretreated Hal31RC. The division index (DI) is given. Pan anti-HLA class I antibody (w6/32) served as specificity control. Data are means of 4 different donors ± SEM. C) Partial reversal of T cell proliferation inhibition upon coculture of CD8⁺ T cells with anti-B7-H1 antibody or respective isotype control preincubated Hal31RC. Hal31RC were pretreated with IL-4 and TNFα for 3 days. Data represent the mean of 3 different donors ± SEM. D) Analyses of constitutive B7H ligand expression, ICOS, CD80 and PD-1 on allogeneic CD8⁺ T cells prior to coculture with RCC31. Data represent means of 7 different donors ± SEM. E) IFNγ production of isolated CD8⁺ T cells upon coculture of cytokine pretreated Hal31RC cells. Data are means of 3 different donors ± SEM.