Journal: Pharmaceutics

Article Title: Curcumin Enhances the Abscopal Effect in Mice with Colorectal Cancer by Acting as an Immunomodulator

doi: 10.3390/pharmaceutics15051519

Figure Lengend Snippet: Biodistribution results of 111 In-anti-OX40 mAb in mice receiving different treatments. Mice were euthanized 48 h after being injected with 111 In-anti-OX40 mAb, designated organs/tissues were collected, and their radioactivity was counted using a gamma counter. Except for blood, the liver, spleen, and tumors showed higher relative radioactivity than other organs. The COMB group had the highest spleen and tumor uptakes among all the groups. a, compared with the CTRL group; b, compared with the Curcumin group; c, compared with the RT group; 1, p < 0.05; 2, p < 0.01.

Article Snippet: Prior to the indium-111 radiolabeling, anti-OX40 mAb (#BE0031, clone OX86, BioXCell, Lebanon, NH, USA) was resuspended in PBS at 0.5 mg/mL and then incubated with fivefold excess DOTA-NHS ester (#HY-128890, MedChemExpress, Monmouth Junction, NJ, USA) at 4 °C overnight.

Techniques: Injection, Radioactivity

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 1. Combined treatment with OX40 agonist and PD1 inhibitor antibodies following VTP therapy suppresses primary tumor growth and improves survival. Percent survival of mice plotted by Kaplan-Meier methods (p < 0.0001).

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques:

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 2. Adding PD-1 inhibitor and OX40 agonist to VTP therapy increases T cell infiltration and activation into MB-49 UTUC allografts at 7 days post-treatment and increases CD4+ and CD8+ T cell infiltration into MB-49 UTUC allografted tumors. (A) Heat map showing the mean frequency of each cell population as measured by flow cytometry in control and treated tumors. Each box represents the mean percentage of positive cells among the parent cell population, i.e., CD45+ among live cells, CD4+ or CD8+ cells among CD45+ cells, Foxp3+ Tregs or Teff among CD4+ cells, CD25+/Foxp3+

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques: Activation Assay, Cytometry, Control

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 3. Adding PD-1 Inhibitor and OX40 agonist to VTP therapy increases T cell infiltration and activation compared to VTP alone, into MB-49 UTUC allografts at 7 days post-treatment. Representative flow cytometry plots showing the percentage of CD4+ and CD8+ cells among CD45+ cells (top row), Ki67+ in CD4+ (second row), Ki67+ in CD8+ (third row), Foxp3+ Tregs and Foxp3−Teff in CD4+ (fourth row), and GrzB+ in CD8+ T cells (bottom row) in control and treated tumors analyzed 7 days after treatment. A fluorescent minus one (FMO) sample was included for each marker as a negative control for gating.

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques: Activation Assay, Flow Cytometry, Control, Marker, Negative Control

Journal: The Journal of Clinical Investigation

Article Title: Imaging activated T cells predicts response to cancer vaccines

doi: 10.1172/JCI98509

Figure Lengend Snippet: (A) Histograms (fluorescence intensity) for OX40, CD44, and CD25 expression following murine T cell activation with PMA/ionomycin or CD3/CD28-specific Ab–coated Dynabeads, and histograms (fluorescence intensity) for human T cells following activation. (B) 3D flow cytometric plots of OX40 expression on resting and activated CD4+ and CD8+ murine T cell subsets. Scale bar: purple = low OX40 expression; yellow = high OX40 expression. (C) Quantitation of OX40 mean fluorescence intensity (MFI) of resting, PMA/ionomycin-stimulated, or Dynabead-activated murine T cells (n = 3 mice/group). (D) Corresponding 64Cu-DOTA-AbOX40 tracer uptake (% dose/105 cells) in resting, PMA/ionomycin-stimulated, or Dynabead-activated murine T cells (n = 6 replicates/group) for unblocked, blocked, and KO groups from 2 independent experiments. (E) 64Cu-DOTA-AbOX40 tracer uptake (% dose/105 cells) in Dynabead-activated murine T cells compared with resting T cells and various murine cancer cell lines (n = 3 mice/group). A20, B cell lymphoma; GL26, glioma; NDL, mammary carcinoma; 4T1, mammary carcinoma; CT26, colorectal carcinoma. All values represent the mean ± SEM unless otherwise specified. ****P < 0.0001, by 2-way ANOVA with Bonferroni’s post test for group comparisons (D) or by 1-way ANOVA (C and E).

Article Snippet: Two types of blocking studies were performed: (a) Anti-mouse OX40 mAb (100 μg; BioXCell) was administered intravenously 30 minutes before radiotracer administration ( n = 3 CpG-treated mice); and (b) a separate cohort of mice ( n = 4 per treatment group) received anti-HRP rat IgG1 isotype control (100 μg; BioXCell) 15 minutes prior to radiotracer administration to further demonstrate specificity of the probe and rule out the contribution of the radiotracer binding to Fc receptors rather than to the OX40 target, since CpG treatment leads to increased Fc receptor expression ( 41 ).

Techniques: Fluorescence, Expressing, Activation Assay, Quantitation Assay

Journal: The Journal of Clinical Investigation

Article Title: Imaging activated T cells predicts response to cancer vaccines

doi: 10.1172/JCI98509

Figure Lengend Snippet: (A) Schematic representation of the in situ vaccination strategy and a hypothetical immune response. (B) Treated tumor (TT) and untreated tumor (UT) growth curves (volume, mm3) based on caliper measurements for CpG-treated (n = 7) and vehicle-treated (n = 8) cohorts. Values represent the mean ± SEM. ****P < 0.0001 and **P < 0.01, by 2-way ANOVA with Bonferroni’s post test. (C) Top: Fold change (normalized to vehicle) in the frequency of OX40+CD3+ T cells in CpG-treated versus untreated TDLNs and (D) tumors at early (n = 5 mice/group) and late (n = 4 mice/group) time points. Bottom: Fold change in the frequency of CD3+ T cells. Gray dashed line demarcates unity. Values represent the mean ± SEM. Statistical analysis was performed using a 2-way ANOVA. P values represent probability differences over time in marker expression due to random chance. (E) Representative viSNE clustering of CD4+, CD8+, and OX40+ markers at the early time point in the CpG-treated cohort. (F) Images of spleens (late time point) from CpG- and vehicle-treated cohorts, presented side by side. Graph shows the fold change (normalized to vehicle) in the frequency of OX40+CD3+ T cells in the CpG-treated cohort at early (n = 5 mice) and late (n = 4 mice) time points. ***P < 0.001, by unpaired Student’s t test. (G) Heatmap of log2 fold change (normalized to control mice) of cytokine expression at both early and late time points in vehicle- and CpG-treated cohorts. Columns represent mice; rows represent cytokines. Scale bars: blue = 0, yellow >4 log2 fold change.

Article Snippet: Two types of blocking studies were performed: (a) Anti-mouse OX40 mAb (100 μg; BioXCell) was administered intravenously 30 minutes before radiotracer administration ( n = 3 CpG-treated mice); and (b) a separate cohort of mice ( n = 4 per treatment group) received anti-HRP rat IgG1 isotype control (100 μg; BioXCell) 15 minutes prior to radiotracer administration to further demonstrate specificity of the probe and rule out the contribution of the radiotracer binding to Fc receptors rather than to the OX40 target, since CpG treatment leads to increased Fc receptor expression ( 41 ).

Techniques: In Situ, Marker, Expressing