CONICET | Buscador de Institutos y Recursos Humanos

Anti-tumor immunity is often impaired by the growing tumor by the induction of immunosuppression and tolerance. Using an experimental murine *brosarcoma (MCC) that progresses from a strongly immunogenic to a tolerogenic state, we have previously demonstrated that MCC growth induces a speci*c immune response that coexists with signs of suppression, like IL-10+ B cells, mainly at the tumor draining lymph node (TDLN). We therefore designed an immunotherapeutic trial consisting of the ablation of TDLN in order to eliminate an incipient focus of suppression, the ex vivo expansion of the ablated TDLN cytotoxic cells with anti-CD3+IL-2, and the re-inoculation of these cells into the donor tumor-bearing mouse by adoptive transference. With this protocol, lower tumor growth, enhanced survival and a high rate of complete tumor remission were obtained. The aim of the present work was to characterize the adoptively transferred cells. Freshly harvested TDLN cells were mostly composed of B, CD4+T and CD8+T cells; ex vivo exposition to anti-CD3+IL-2 increased CD4+ and CD8+T and decreased B cell proportions (% cells before versus after culture: TCD4+: 37±1 vs 74±7***; TCD8+: 17.3±0.5 vs 26.8±6**; B220+CD19+: 44±3 vs 7.3±2***, n=6), and increased the intracellular IFN-#- expressing T cells (% IFN-#+/ CD4+ cells before versus after the culture: 4.3±1.7 vs 12.0±4.6*; % IFN-#+/CD8+: 8.9±5.4 vs 20.8±6.8*, n=5). Additionally, cultured cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). CD4+ cells before versus after the culture: 4.3±1.7 vs 12.0±4.6*; % IFN-#+/CD8+: 8.9±5.4 vs 20.8±6.8*, n=5). Additionally, cultured cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). #- expressing T cells (% IFN-#+/ CD4+ cells before versus after the culture: 4.3±1.7 vs 12.0±4.6*; % IFN-#+/CD8+: 8.9±5.4 vs 20.8±6.8*, n=5). Additionally, cultured cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). #+/CD8+: 8.9±5.4 vs 20.8±6.8*, n=5). Additionally, cultured cells showed enhanced anti-MCC cytotoxicity evaluated by JAM test. By staining cells with CFSE prior to their adoptive transference, we were able to detect them within the tumor tissue, spleen and distant lymph node on day 7 after the innoculum. Our results suggest that the anti-tumor e´ects obtained with the treatment are due, at least in part, to anti-CD3+IL-2- induced decrease of B cells and increase of IFN#+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001). #+ T cells with enhanced cytotoxic capacity. (*p<0.05, **p<0.01, ***p<0.001).