Figure 3.

In situ Flagrp170 immunotherapy induces GM-CSF-dependent antitumor response against mouse mammary tumor. (A) BALB/c mice (n=5) with established 4T1 tumors of 4~5 mm sizes were treated with an empty or Flagrp170-encoding adenovirus as indicated. GM-CSF antibodies (200 µg) was administrated intraperitonially 1 day before the first treatment and continued at 3-day intervals for four doses. (B) Gene transcription of ifng, il12a, and gmcsf in tumor tissues (n=3) was assayed 2 weeks after the Flagrp170 treatment. (C) Expression of IFN-γ and granzyme B in tumor-infiltrating CD8⁺ T cells or NK cells were examined by intracellular cytokine staining. Splenocytes (D) or draining lymph node cells (E) from treated mice (n=3) were stimulated with 4T1 cell lysates. Production of cytokines IFN-γ and IL-2 was assessed using ELISA. Data shown are representative of three independent experiments. *p<0.05, **p<0.01, ***p<0.001, using two-way repeated measures analysis of variance test (A) and Student’s t-test (B, D, E). GM-CSF, granulocyte macrophage colony-stimulating factor; IFN, interferon; IL, interleukin.