Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: Comparison of Antitumor Effects of Combinations of Immune Checkpoint Inhibitors With Dendritic Cells Intratumorally Injected into Irradiated Mouse Adenocarcinoma

doi: 10.1097/CJI.0000000000000548

Figure Lengend Snippet: Schematic diagram showing the mouse treatment protocol. MC-38 cells were inoculated subcutaneously into the right thigh (8×10 5 cells) and left flank (4×10 5 cells) of C57BL/6 mice. When the thigh tumor size reached 8×8 mm, on day 12 after tumor inoculation, a first dose of 10 Gy radiation was administered to the right thigh, and iDCs (1×10 6 cells) were injected intratumorally 24 hours after irradiation. A second dose of 10 Gy radiation was irradiated on day 19 after tumor inoculation, and secondary iDCs were injected intratumorally 24 hours after a second irradiation. Antibodies against programmed cell death-1 (10 mg/kg) and programmed cell death-ligand 1 (10 mg/kg) were administered intraperitoneally 3 times per week for 2 weeks. Saline was administered to the control groups in the same manner. Mice were sacrificed 48 hours after the last antibody treatment. iDC indicates immature dendritic cells.

Article Snippet: Anti-PD-L1 antibody (R&D Systems Inc.; cat. no. MAB90781) was diluted 1:200 and incubated at 4°C overnight.

Techniques: Injection, Irradiation, Saline, Control

Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: Comparison of Antitumor Effects of Combinations of Immune Checkpoint Inhibitors With Dendritic Cells Intratumorally Injected into Irradiated Mouse Adenocarcinoma

doi: 10.1097/CJI.0000000000000548

Figure Lengend Snippet: PD-L1 expression of iDCs and distant tumor. (A) PD-L1 expression on the surface of iDCs was evaluated using flow cytometry FC500 at weeks 1 and 2 of iDCs administration. The overlay graph shows the expression of PD-L1 (red) along with the corresponding isotype control (green). (B) Immunohistochemistry staining of mouse flank tumor. Each photomicrograph shows DAB staining (brown) for PD-L1 and hematoxylin staining (blue) for nuclei in tumor sections (×400 magnification). Figures are representative of 5 mice/group. iDC indicates immature dendritic cells; PD-L1, programmed cell death-ligand 1.

Article Snippet: Anti-PD-L1 antibody (R&D Systems Inc.; cat. no. MAB90781) was diluted 1:200 and incubated at 4°C overnight.

Techniques: Expressing, Flow Cytometry, Control, Immunohistochemistry, Staining

Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: Comparison of Antitumor Effects of Combinations of Immune Checkpoint Inhibitors With Dendritic Cells Intratumorally Injected into Irradiated Mouse Adenocarcinoma

doi: 10.1097/CJI.0000000000000548

Figure Lengend Snippet: Tumor volume of tumor-bearing mice. Tumor sizes in the (A–D) right thigh and (E–H) left flank were measured by calipers. (B and F) The bar graphs in each panel show the tumor size measured on day 24, which is 24 hours after the final antibody administration (n=5–6). The graphs in panels (A) and (E) were separated into distinct figures for (C, G) aPD-1-related groups and (D, H) aPD-L1-related groups, respectively. Tumor volume was calculated using the modified ellipsoid formula “length × width 2 × 0.5.” Statistical significance was determined using the Brown-Forsythe and Welch ANOVA test (right thigh tumor) and 1-way ANOVA with the Tukey post hoc test (left flank tumor). * P <0.05, ** P <0.01, **** P <0.0001.

Article Snippet: Anti-PD-L1 antibody (R&D Systems Inc.; cat. no. MAB90781) was diluted 1:200 and incubated at 4°C overnight.

Techniques: Modification

Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: Comparison of Antitumor Effects of Combinations of Immune Checkpoint Inhibitors With Dendritic Cells Intratumorally Injected into Irradiated Mouse Adenocarcinoma

doi: 10.1097/CJI.0000000000000548

Figure Lengend Snippet: Kaplan-Meier survival curves and median survival time after combination treatment of irradiation/immature dendritic cells and immunotherapy (anti-programmed cell death-1 or anti-programmed cell death ligand-1; n=5). Statistical significance was determined using the log-rank test. * P <0.05, ** P <0.01.

Article Snippet: Anti-PD-L1 antibody (R&D Systems Inc.; cat. no. MAB90781) was diluted 1:200 and incubated at 4°C overnight.

Techniques: Irradiation