Journal: Oncoimmunology

Article Title: Timing of CSF-1/CSF-1R signaling blockade is critical to improving responses to CTLA-4 based immunotherapy

doi: 10.1080/2162402X.2016.1151595

Figure Lengend Snippet: Blockade of CSF-1R signaling reprograms the tumor microenvironment (A) Mice were injected with 4T1 tumor cells and treated with αCSF-1R or IgG. Frequencies of CD11b+Ly6G−/lowLy6Chigh. MO-MDSCs, CD11b+Ly6G+Ly6C−/low G-MDSCs and total CD11b+ myeloid cells of total CD45+ cells in tumors after three and six doses of treatment. (B) Expression of CSF-1R within the CD11b+Ly6G−/lowLy6Chigh and CD11b+Ly6G+Ly6C−/low gated populations. (C) CD11b+Ly6G−/lowLy6Chigh and CD11b+Ly6G+Ly6C−/low subsets from 4T1 tumors were evaluated for expression of CD11c, MHC class II and F4/80 markers (filled histograms) against their matched isotype controls (open histograms). Data are mean +/– SEM.

Article Snippet: One day later, cells were collected and stained with anti-CD14-eFlour450 (clone M5E2), anti-HLA-ABC-FITC, anti-CSF-1R-PE (Clone 9-4D2-1E4), anti-HLA-DR-PE Texas Red (clone G46-6), anti-CD11b-PerCP-Cy5.5 (clone M1/70), anti-CD33-PE-Cy7 (clone P67.6), and anti-CD45-APC eFluor 780 (clone 2D1) antibodies, all purchased from BD Biosciences, eBioscience or Invitrogen.

Techniques: Injection, Expressing

Journal: Oncoimmunology

Article Title: Timing of CSF-1/CSF-1R signaling blockade is critical to improving responses to CTLA-4 based immunotherapy

doi: 10.1080/2162402X.2016.1151595

Figure Lengend Snippet: The combination of αCTLA-4 and αCSF-1R modifies the tumor microenvironment and increases intra tumor inflammation. (A) 4T1 tumors were harvested on 15 (after treatment with αCSF-1R and/or αCTLA-4) and 23 (after sequential or concurrent treatment with αCSF-1R and αCTLA-4) post-tumor inoculation and analyzed for tumor-infiltrating immune cells. Treatment schedule for each group: Group A) IgG, Group B) Sequential αCSF-1R and αCTLA-4, Group C) Sequential αCTLA-4 and αCSF-1R, and Group D) Concurrent αCSF-1R + αCTLA-4. (B) Results for day 15: Frequency of CD11b+Ly6G−/lowLy6Chigh. MO-MDSCs of total CD45+ cells and representative dot plots, mean tumor weight, and absolute number of CD8+ T cells and CD4+ Teff cells per gram of tumor. (C) Results for day 23: Frequency of CD11b+Ly6G−/lowLy6Chigh. MO-MDSCs of total CD45+ cells, mean tumor weight, and absolute number of CD8+ T cells and CD4+ Teff cells per gram of tumor. (D–E) Data is derived from nanoString analysis using the 750 gene Mouse Immunology Panel. Differential gene expression using the Broad GenePattern “Comparative Marker Selection” tool identified a set of 200 genes down-regulated in CS7 versus control with p value < 0.05. These 200 were removed, leaving 550 genes, which were reanalyzed in Comparative Marker Selection using “one-versus-all” as a comparison mode. From this a total of 64 genes were found to be differentially expressed at the p value < 0.05 level, of which two were shared with CS7 and two with CTLA4, leaving 60 unique genes differentially expressed versus the aggregate of control, CS7, and CTLA4. Data are mean +/– SEM.

Article Snippet: One day later, cells were collected and stained with anti-CD14-eFlour450 (clone M5E2), anti-HLA-ABC-FITC, anti-CSF-1R-PE (Clone 9-4D2-1E4), anti-HLA-DR-PE Texas Red (clone G46-6), anti-CD11b-PerCP-Cy5.5 (clone M1/70), anti-CD33-PE-Cy7 (clone P67.6), and anti-CD45-APC eFluor 780 (clone 2D1) antibodies, all purchased from BD Biosciences, eBioscience or Invitrogen.

Techniques: Derivative Assay, Expressing, Marker, Selection