Functional and molecular characterization of interleukin-2 transgenic Ewing tumor cells for in vivo immunotherapy

Background. Interleukin-2 (IL-2) is a potent cytokine with potential activity against several tumors including Ewing tumors (ET). Side effects of systemic IL-2 can be circumvented by the use of transgenic tumor cells. However, in vitro manipulation may change the overall gene expression profile of tumor cells unfavorably. Therefore, we assessed gene expression profiles, safety, and immunomodulatory efficacy of IL-2 transgenic (IL-2-tg) ET cells in vitro and in NOD/ scid mice. Procedure. Viable wild type A673 tumor cells were co-cultured together with irradiated IL-2-tg or mock-transfected cells and HLA matched peripheral blood mononuclear cells. Activation of T and NK cells was assessed by FACS analysis. The effect of irradiated IL-2-tg cells on tumor growth in vivo was investigated by using NOD/scid mice. Gene expression profiles of wild type and transfected cells were analyzed with Affymetrix HG-U95A microarrays. Results. IL-2-tg cells activated and increased the number of T cells and NK cells in vitro. Co-culture with IL-2-tg but not with mock-transfected cells almost completely suppressed wild type tumor cell growth in vitro. Cell depletion experiments indicated a major contribution of NK cells to this tumor cell suppression. Co-transfer of irradiated IL-2-tg cells significantly reduced wild type tumor growth in NOD/scid mice. Side effects in the treated animals were not observed and no tumor growth was observed after injection of irradiated IL-2-tg cells alone. Gene expression profiling revealed a substantial degree of homogeneity of gene expression in transfected and wild type cells and suggests that transfection and selection procedures had no major impact on the gene expression profile. Conclusions. Next to a high degree of homogeneity between transgenic and wild type cells, our data suggest that irradiated IL-2-tg ET cells can activate cytolytic effector cells. These cells may have therapeutic potential for ET patients.