Generation Of Specific Polyclonal And Polyfunctional Cd4(+) T-Helper1 Cells Against Wt-1, Mage-A3, Survivin And Ror-1 For Adoptive T-Cell Immunotherapy.

e14025 Background: Adoptive T cell therapy (ACT) is a promising option to treat cancer. Th1 cells have been shown effective against viremia and cancer and are mediators of senescence in tumor cells. T cells against tumor associated antigens like cancer testis antigens can be used for adoptive T-cell transfer approaches. Methods: According to these considerations, we developed a protocol according to good manufacturing practice (GMP) to generate Th1 cells against WT-1, MAGE-A3, Survivin and ROR-1. The protocol starts with a presensitization of T cells with antigen. T cells were then enriched based on IFN-γ capture technique and expanded using autologous feeder and IL-2, IL-7 and IL-15. T-cell specificity and function was analyzed by flow cytometry. Results: Large scale generation of T cells specific to the tumor antigens was performed. The T-cell lines showed high numbers of specific IFN-γ+ and TNF-α+ CD4+ T cells but no IL-10. Both, CD4+ and CD8+T cells had an effector memory phenotype and proliferated in response to the tumor antigens. Adding PD-1 antibody to the T cells resulted in an improved proliferation in response to the antigens. Treatment of a WT-1+ leukemia patient with stem cell donor derived WT-1 specific T cells in combination with WT-1 vaccine was accompanied with WT-1 CD4+ and CD8+T cell responses and ongoing remission. Conclusions: Protocols for the generation of Survivin, WT-1-, MAGE-A3- and ROR-1-specific T cells were successfully established. T cell products mainly consisted of CD4+ T-cells with secretion of the cytokines IFN-γ and TNF. This cytokine profile enables the induction of senescence in tumor cells. Although tumor associated antigens are potential self antigens, it is possible to induce a functional Th1 response in T cells from healthy donors making T-cell transfer feasible in an allogeneic settings post stem cell transplantation. Further improvements could be achieved by combination of T-cell therapy with antibodies to PD-1.