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The vaccine consisted of 106 -irradiated CT26 or C26GM cells injected s.c. For ACT experiments, mice were vaccinated on days –14 and –7 before ACT in all four limbs with 106 -irradiated C26GM. The CD8+ T cells were negatively purified using the CD8+ isolation kit (Miltenyi Biotec), and 3 x 106 CD8+ T cells were transferred per mouse. In the 4T1-HA experiments, HA-specific CD8+ naive T cells were isolated from spleens and lymph nodes of pCL4-TCR mice.
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"There is an association between these two conditions, but we still don't know the exact mechanism," Gao said. His team said further research is warranted.
"If a man says 'no' to you with a lot of irritability, I still wouldn't take it personally or feel rejected, because it's very likely he's suffering some depression -- and again, it's not you," says Foley.
Although the hallmark feature of MDSCs is immunosuppression, emerging data reveal that the degree of immunosuppression varies among populations of MDSCs isolated from different organs, with intratumoral MDSCs being the most immunosuppressive. Interestingly, these MDSCs express greater levels of NOS2 and ARG1 than their splenic counterparts (5). ARG1 expression is mainly regulated by the STAT-6–IL-4R pathway (30). We recently correlated IL-4R expression on CD11b+/Gr-1+ with an immunosuppressive phenotype (29), and our in vitro data (Fig. 5) indicate that sildenafil down-regulates IL-4R on MDSCs. We then asked whether in vivo PDE5 inhibition reduced ARG1 and NOS2 and down-regulated IL-4R in tumor-associated MDSCs. BALB/c mice were challenged with CT26WT, and half were treated with sildenafil. Mice were killed 15 d later, and intratumoral MDSCs were obtained. Sildenafil increased cGMP (Fig. 6 A), reduced IL-4R expression (Fig. 6 B), and down-regulated NOS2 and ARG1 expression and reduced their enzymatic activity in the intratumoral MDSCs (Fig. 6, C and D). Considering that ARG1 and NOS2 are key enzymes in MDSC suppressive pathways (8, 31), these findings support the hypothesis that PDE5 inhibition is a novel pharmacologic approach to regulate MDSC-mediated immunosuppressive pathways.
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