Immunotherapy against breast cancer targeting Stat3 triggers NK/CD4+ T-cell mediated antitumoral response and inhibits tumor growth in therapeutic administration Tumor Immunology

TKACH M; CORIA L; ROSEMBLIT C; RIVAS MA; BEGUELIN W; PROIETTI CJ; DIAZ FALQUE MC; CHARREAU EH; ELIZALDE PV; CASSATARO J; SCHILLACI R

Miami, Florida

Congreso; Tumor Immunology: Basic and Clinical advances;; 2010

American Association for Cancer Research (AACR)

The evidence that disruption of Stat3 signaling in cancer cells can overcome tumor immune evasion led us to design an immunotherapy based on the administration of irradiated breast cancer cells that express a dominant negative (DN) form of Stat3. We have already shown that this immunization provides protection against the murine progestin-dependent C4HD tumor in a prophylactic protocol. On the one hand, now our focus is disclosing the lymphocyte subsets involved in the antitumoral response and on the other, evaluating the therapeutic effect of DNStat3 C4HD cells against tumor progression. To achieve our first objective, immunized BALB/c mice were in vivoin vivo depleted of CD4+, CD8+, or NK cells with monoclonal antibodies before tumor challenge. Depletion of CD4+ T or NK cells completely abrogated resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a tumor challenge. Depletion of CD4+ T or NK cells completely abrogated resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a tumor challenge. Depletion of CD4+ T or NK cells completely abrogated resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a +, CD8+, or NK cells with monoclonal antibodies before tumor challenge. Depletion of CD4+ T or NK cells completely abrogated resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a + T or NK cells completely abrogated resistance to tumor challenge induced by immunization with DNStat3- transfected C4HD cells (*p<0,01 and **p<0,001, respectively). However, depletion of CD8+ T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a + T cells did not affect C4HD tumor rejection. We demonstrated that splenocytes from mice injected with DNStat3-C4HD cells were effective in lysing C4HD in vitro. To determine whether NK cells were responsible for that cytotoxic effect, we isolated them from spleens of immunized mice and performed a 51Cr release assay. NK cells from mice immunized with DNStat3- transfected C4HD cells showed an increased cytotoxicity against YAC- 1 cell line and C4HD cells compared to mice immunized with empty vector-transfected C4HD cells (27,5 ± 2,5% vs 5,2 ± 1,9% and 15,7± 2,4 vs 5,3± 2,7, respectively, 5:1 effector-to-target ratio, p<0,05). We also identified the cell subsets responsible for this cytotoxic effect in vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ transfected C4HD cells showed an increased cytotoxicity against YAC- 1 cell line and C4HD cells compared to mice immunized with empty vector-transfected C4HD cells (27,5 ± 2,5% vs 5,2 ± 1,9% and 15,7± 2,4 vs 5,3± 2,7, respectively, 5:1 effector-to-target ratio, p<0,05). We also identified the cell subsets responsible for this cytotoxic effect in vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ transfected C4HD cells showed an increased cytotoxicity against YAC- 1 cell line and C4HD cells compared to mice immunized with empty vector-transfected C4HD cells (27,5 ± 2,5% vs 5,2 ± 1,9% and 15,7± 2,4 vs 5,3± 2,7, respectively, 5:1 effector-to-target ratio, p<0,05). We also identified the cell subsets responsible for this cytotoxic effect in vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ vitro by depleting NK cells, CD4+ or CD8+ T cell populations from mouse splenocytes by immunomagnetic separation before performing the 51Cr release assay. Depletion of NK cells from splenocytes of mice immunized with DNStat3-transfected C4HD cells resulted in total suppression of cytotoxic activity. In contrast, depletion of CD4+ or CD8+ immunized with DNStat3-transfected C4HD cells MENÚ Acerca del CONICET Autoridades Normativa Contacto Inicio Evaluación Convocatorias Red institucional Vinculación tecnológica Proyectos de investigación Comunicación Concursos y premios Contacto Términos y condiciones Consejo Nacional de Investigaciones Científicas y Técnicas. Av. Rivadavia 1917 (C1033AAJ) Ciudad Autónoma de Buenos Aires - República Argentina Tel: +5411 5983-1420 - +5411 4953-7230 www.conicet.gov.ar