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. 2012 Nov 13;21(1):167–174. doi: 10.1038/mt.2012.225

Figure 5.

Figure 5

Regulated interleukin (IL)2 activity enhanced the tumor-targeted immune response. Tumor and spleen were collected from female BALB/c mice bearing subcutaneous RENCA tumors 7 days after animals were injected systemically with PBS or 3 × 108 PFU of vaccinia virus double deleted (vvDD) or vvDD-L106P-IL2. Shield-1 was used to stabilize IL2 function immediately after the administration of virus or 72 hours after the administration of virus as before. (a) Splenic lymphocytes were stained using antibodies against CD3, CD4, CD8, Ki-67, and pS6. Levels of activated CD4+ and CD8+ T-cells increased for treatments with both constitutive (P = 0.019 and 0.017) and regulated IL2 (P = 0.018 and 0.013) relative to vvDD alone. (b) Tumor-infiltrating T-cell lymphocytes were stained using antibodies against CD3, CD4, and CD8. Constitutive IL2 expression did not significantly increase tumor levels of either CD8+ or CD4+ relative to vvDD (P = 0.99 and 1.02), whereas regulated IL2 significantly increased the levels of CD8+ T-cells (P = 0.049), while CD4+ increases trended higher (P = 0.054) n = 3. (c) Tumor-infiltrating regulatory T-cells after treatment with different viral strains. Tumors as before were also stained for T-reg after permeabilization of dissociated cells (CD4+CD25+FoxP3+), levels of T-reg in the tumors are shown. PBS, phosphate-buffered saline.