Journal: Cancers

Article Title: Targeting Proliferating Tumor-Infiltrating Macrophages Facilitates Spatial Redistribution of CD8 + T Cells in Pancreatic Cancer

doi: 10.3390/cancers14061474

Figure Lengend Snippet: Proliferating F4/80 + macrophages were reduced in Panc02 and KPC mouse models after clodronate liposomes treatment. ( A , B ) Representative immunofluorescence images of Ki-67 and F4/80 staining in Panc02 or KPC tumors and quantitation of Ki-67 and F4/80 immunofluorescence staining. ( C ) Analysis of BrdU + macrophages in Panc02 and KPC tumor tissues. The animals were injected with BrdU at 12 h and 24 h before sacrifice. ( D , E ) Staining of CD8 + T and Foxp3 + T cells in Panc02 or KPC tumors with or without clodronate liposomes treatment. Data are shown as the mean ± SEM. Statistical analyses were carried out by Student’s t test. *: p ≤ 0.05, **: p ≤ 0.01.

Article Snippet: The Abs used were purchased from Cell Signaling Technology: anti-CD68 (D4B9C), anti-CD86 (E2G8P), anti-CD163 (D6U1J), anti-pan-keratin (C11), anti-F4/80 (D2S9R), anti-Ki-67 (D3B5), and anti-CD8 (D4W2Z); an Opal 4-color Manual IHC Kit (PerkinElmer, Spokane, WA, USA) was used for mIHC.

Techniques: Immunofluorescence, Staining, Quantitation Assay, Injection

Journal: Cancers

Article Title: Targeting Proliferating Tumor-Infiltrating Macrophages Facilitates Spatial Redistribution of CD8 + T Cells in Pancreatic Cancer

doi: 10.3390/cancers14061474

Figure Lengend Snippet: The antitumor response induced by targeting macrophages is CD8 + T-cell-dependent. ( A ) Representative flow cytometric analysis of CD8 + T cells to confirm depletion was carried out. ( B , C ) Mice were treated with 200 μL (intraperitoneal injection) of control liposomes or clodronate liposomes on days −1 and 0 and every 3 days afterward, and/or with isotype control or anti-CD8 antibodies on days −1, 0, 5, 10, and 15 ( n = 5 mice/group). Left: tumor growth curves showing the volumes of subcutaneously implanted tumors in mice ( n = 5 per group) treated with control liposomes or clodronate liposomes. Right: final tumor weights. ( D , E ) Flow cytometric analysis of tumor-infiltrating CD8 + T cells ( D ) and Foxp3 + Tregs ( E ). Each data point represents a mouse. Data are shown as the mean ± SEM. Statistical analyses were performed by one-way ANOVA with Tukey’s correction for multiple comparisons. *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001.

Article Snippet: The Abs used were purchased from Cell Signaling Technology: anti-CD68 (D4B9C), anti-CD86 (E2G8P), anti-CD163 (D6U1J), anti-pan-keratin (C11), anti-F4/80 (D2S9R), anti-Ki-67 (D3B5), and anti-CD8 (D4W2Z); an Opal 4-color Manual IHC Kit (PerkinElmer, Spokane, WA, USA) was used for mIHC.

Techniques: Injection

Journal: Cancers

Article Title: Targeting Proliferating Tumor-Infiltrating Macrophages Facilitates Spatial Redistribution of CD8 + T Cells in Pancreatic Cancer

doi: 10.3390/cancers14061474

Figure Lengend Snippet: Targeting macrophages promoted CD8 + T cell spatial redistribution in tumors. Panc02 and KPC murine pancreatic tumor cells were subcutaneously injected into C57BL/6 mice, and the effects of treatment with control liposomes or clodronate liposomes on intratumoral T cell infiltration and spatial redistribution after macrophage targeting were evaluated. ( A , B ) Representative images of the edge (periphery) and center (core) of the tumor sections after anti-CD8 immunostaining are shown. For each tumor, the numbers of CD8 + T cells at the edge of the tumor (≤500 µm from the edge of the tumor) and the tumor center (>500 μm from the edge of the tumor) were quantified. The distance between the nuclear centers of a PanCK + tumor cells and their nearest CD8 + T cells was calculated for each PanCK + cell identified within the tumor center. ( C , D ) The average distances of CD8 + T cells to tumor cells in all tumor samples were calculated. ( C , D ) ( right ) The distribution of CD8 + T cells within 30 μm from tumor cells. Each group contained five mice, corresponding to the five points in the bar graph. We analyzed at least five views for each point and calculated the average density of cells at the center and edge to represent each slide. ( E ) Representative flow cytometric analysis of IFN-γ + CD8 + cells in CD8 + T cells and a statistical graph are shown. Data are shown as the mean ± SEM. Statistical analyses were carried out by Student’s t test. *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, ns: not significant ( p > 0.05).

Article Snippet: The Abs used were purchased from Cell Signaling Technology: anti-CD68 (D4B9C), anti-CD86 (E2G8P), anti-CD163 (D6U1J), anti-pan-keratin (C11), anti-F4/80 (D2S9R), anti-Ki-67 (D3B5), and anti-CD8 (D4W2Z); an Opal 4-color Manual IHC Kit (PerkinElmer, Spokane, WA, USA) was used for mIHC.

Techniques: Injection, Immunostaining