Figure 7. Treatment with a neutralizing monoclonal antibody against IL-17RB blocks tumor growth and metastasis, and extends survivals in a mouse orthotopic xenografted model. (A and B) Soft agar colony formation (A) and invasion (B) assays using CFPAC-1 and BxPC3 cells treated with control IgG or D9 antibody. All experimental data were verified in at least two independent experiments. (C) Relative sensitivity of both control and IL-17RB–KD clones to IgG and anti–IL-17RB in colony formation activities. (D) Schematic diagram of antibody treatment in orthotopically xenografted mice. (E and F) IVIS image (E) and tumor weight (F) of antibody-treated NOD/SCIDγ mice orthotopically implanted with CFPAC-1 cells on day 28. Cell dose: 2.5 × 105 cells per mouse. Eight mice were used for each group. Two mice from each group were sacrificed on day 28 for measurement of tumor weight and lung metastasis. (G) Lung metastasis of antibody-treated NOD/SCIDγ mice orthotopically injected with CFPAC-1 cells. Cell dose: 2.5 × 105 cells per mouse. Six mice were used for each group. (H) Immunoblot analysis of IL-17RB using tumors derived from antibody-treated mice xenografted with CFPAC-1 cells. GAPDH was used as a loading control. All experimental data verified in at least two independent experiments. (I) Representative pictures of the IHC analyses of IL-17RB in tumors from control IgG or mAb-treated xenografted mice. Bar, 100 µm. Boxes show the enlarged area. (J) mRNA expressions of CCL20, CXCL1, IL-8, and TFF1 were measured by qRT-PCR in tumors derived from antibody-treated mice xenografted with CFPAC-1 cells. β-actin was used as an internal control. Relative expression of chemokines in D9-treated cells relative to control IgG-treated cells is shown. (K) Representative pictures of the IHC analyses of TFF-1 in tumors from control IgG or mAb-treated xenografted mice. Bar, 100 µm. Boxes show the enlarged area. All experimental data verified in at least two independent experiments. (L) Comparison of the survival periods of the antibody-treated NOD/SCIDγ mice orthotopically injected with CFPAC-1 cells using the Kaplan-Meier method. Data shown are means ± SD. P < 0.001. All xenograft experiments were performed once with CFPAC-1 cells and similar results were observed using BxPC3 cells (not depicted).
Figure 7.

Treatment with a neutralizing monoclonal antibody against IL-17RB blocks tumor growth and metastasis, and extends survivals in a mouse orthotopic xenografted model. (A and B) Soft agar colony formation (A) and invasion (B) assays using CFPAC-1 and BxPC3 cells treated with control IgG or D9 antibody. All experimental data were verified in at least two independent experiments. (C) Relative sensitivity of both control and IL-17RB–KD clones to IgG and anti–IL-17RB in colony formation activities. (D) Schematic diagram of antibody treatment in orthotopically xenografted mice. (E and F) IVIS image (E) and tumor weight (F) of antibody-treated NOD/SCIDγ mice orthotopically implanted with CFPAC-1 cells on day 28. Cell dose: 2.5 × 105 cells per mouse. Eight mice were used for each group. Two mice from each group were sacrificed on day 28 for measurement of tumor weight and lung metastasis. (G) Lung metastasis of antibody-treated NOD/SCIDγ mice orthotopically injected with CFPAC-1 cells. Cell dose: 2.5 × 105 cells per mouse. Six mice were used for each group. (H) Immunoblot analysis of IL-17RB using tumors derived from antibody-treated mice xenografted with CFPAC-1 cells. GAPDH was used as a loading control. All experimental data verified in at least two independent experiments. (I) Representative pictures of the IHC analyses of IL-17RB in tumors from control IgG or mAb-treated xenografted mice. Bar, 100 µm. Boxes show the enlarged area. (J) mRNA expressions of CCL20, CXCL1, IL-8, and TFF1 were measured by qRT-PCR in tumors derived from antibody-treated mice xenografted with CFPAC-1 cells. β-actin was used as an internal control. Relative expression of chemokines in D9-treated cells relative to control IgG-treated cells is shown. (K) Representative pictures of the IHC analyses of TFF-1 in tumors from control IgG or mAb-treated xenografted mice. Bar, 100 µm. Boxes show the enlarged area. All experimental data verified in at least two independent experiments. (L) Comparison of the survival periods of the antibody-treated NOD/SCIDγ mice orthotopically injected with CFPAC-1 cells using the Kaplan-Meier method. Data shown are means ± SD. P < 0.001. All xenograft experiments were performed once with CFPAC-1 cells and similar results were observed using BxPC3 cells (not depicted).

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