Journal: Journal for Immunotherapy of Cancer

Article Title: Brain tumors induce immunoregulatory dendritic cells in draining lymph nodes that can be targeted by OX40 agonist treatment

doi: 10.1136/jitc-2025-011548

Figure Lengend Snippet: Treatment with an anti-OX40 agonist enhances immune responses in cervical TdLN. Schematic illustration of the experimental layout ( A ) used to obtain data shown in ( B–F ): groups of 5–7 mice were inoculated with the glioma cell line SB28-OVA, followed by i.p. administration of either an anti-OX40 agonist or IgG control on days 4, 7, and 10 post-tumor inoculation. On day 12 post-tumor inoculation, the TdLN-c were harvested, and single-cell suspensions were collected for immune profile analysis using flow cytometry. ( B ) H2b-SIINFEKL-positive DCs in TdLN on day 12 after treatment with OX40 agonist or control antibody. ( C ) Activation markers, CD40 and CD86, expressed by DCs. ( D ) Regulatory T cells (CD4+CD25+ Foxp3+). ( E ) Regulatory T cell (CD127+/-). ( F ) Memory T-cell response for CD4+ and CD8+ central memory (CM, CD44+CD62L+) and effector memory (EFM, CD44+CD62L-). ( G, I ) Tumor growth was evaluated using bioluminescence imaging. ( H ) H&E staining of brain tissue from the endpoint. ( J ) Kaplan–Meier survival curves of tumor-bearing mice treated with OX40 agonist or control IgG. The graph shows combined survival curves obtained from two independent experiments. Bar graphs show the mean±SD. One-way ANOVA multiple comparisons test *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; i.p., intraperitoneally; TdLN, tumor-draining lymph nodes.

Article Snippet: The following stimuli were added: Poly-I:C 5 μg/mL (Adipogen Life Sciences), αCD40 (FGK45), αIL-6 (MP5-20F3), αIL-4 (BVD6-24G2), αOX40 (OX-86), αCD200 (OX-90), and αPDL-1 (10F.9G2TM) (BioXcell) at a concentration of 100 ng/mL.

Techniques: Control, Flow Cytometry, Activation Assay, Imaging, Staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Brain tumors induce immunoregulatory dendritic cells in draining lymph nodes that can be targeted by OX40 agonist treatment

doi: 10.1136/jitc-2025-011548

Figure Lengend Snippet: CCR7+OX40+ dendritic cells (DCs) in cervical lymph nodes correlate with anti-OX40 agonist immunotherapy. DCs from TdLNs were isolated and cultured in the presence of the labeled peptide DQ-OVA, which allowed for quantification of antigen uptake, processing, and presentation. (A, B) Representative histograms and percentages of DQ-OVA signals in different DC populations. (C) DCs from TdLN were cultured as in (A) and incubated in the presence of anti-OX40 agonist, Poly-I:C or IgG control. The percentages of CD40 (C), CD86 (D), and PDL-1 (E) were calculated. (F) The percentage of CCR7+OX40+ DCs was calculated from TdLNs harvested 12 days post-tumor inoculation. (G) Representative immunofluorescence staining images of brain sections showing LAMP3+ (*), CD4+ (+), and OX40+ (¤). (H) The graph shows the average number of positive cells across different regions of interest (ROIs, n=392). (I, J) Kaplan-Meier survival curves of EG7-OVA tumor-bearing mice treated with agonistic OX40 antibody or IgG control, either alone or combined with FTY720, were analyzed using the log-rank test (n=5–7 mice/group test). Statistical analyses were conducted using the Mann-Whitney U test. Bar graphs show the mean±SD; two-way ANOVA with Tukey’s correction: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance TdLNs, tumor-draining lymph nodes.

Article Snippet: The following stimuli were added: Poly-I:C 5 μg/mL (Adipogen Life Sciences), αCD40 (FGK45), αIL-6 (MP5-20F3), αIL-4 (BVD6-24G2), αOX40 (OX-86), αCD200 (OX-90), and αPDL-1 (10F.9G2TM) (BioXcell) at a concentration of 100 ng/mL.

Techniques: Isolation, Cell Culture, Labeling, Incubation, Control, Immunofluorescence, Staining, MANN-WHITNEY

Journal: Cancer research

Article Title: Mucosal HPV E6/E7 Peptide Vaccination in Combination with Immune Checkpoint Modulation Induces Regression of HPV + Oral Cancers

doi: 10.1158/0008-5472.CAN-18-0892

Figure Lengend Snippet: Intranasal HPV peptide vaccination enhances the therapeutic efficacy of α4–1BB and αCD40 antibodies. A, Mice were injected subcutaneously with mEER cells (1 × 106) and when the tumors were palpable, received intranasal HPV peptide vaccination on days 6 and 12 along with intraperitoneal injections of α4–1BB, αCD40, αCTLA-4, αPD-1, or αOX40 at days 6, 9, and 12. The tumor growth (B) and survival (C) were monitored over time. The percentage of mice showing tumor regression is noted for each treatment (B). A Mantel–Cox test was performed to determine the significance of survival for each of the treatment groups relative to untreated group; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005. Results represent pooled data from multiple experiments (n = 7–27 mice/group).

Article Snippet: The following antibodies for immunotherapy were purchased from BioXcell and used at the concentrations shown: α4–1BB (LOB12.3 at 350 μg per dose), αCD40 (FGK4.5 at 100 μg per dose), αCTLA-4 (9H10 at 100 μg per dose), αPD-1 (RMP1–14 at 250 μg per dose), and αOX-40 (OX-86 at 100 μg per dose).

Techniques: Injection

Journal: Cancer research

Article Title: Mucosal HPV E6/E7 Peptide Vaccination in Combination with Immune Checkpoint Modulation Induces Regression of HPV + Oral Cancers

doi: 10.1158/0008-5472.CAN-18-0892

Figure Lengend Snippet: Combination treatment consisting of α4–1BB and αCTLA-4 in the presence and absence of HPV peptide vaccine does not exhibit liver toxicities. Serum samples from mice treated with different vaccine and immunotherapy combinations were collected at day 14 after tumor challenge and analyzed for AST (A; n = 5–16 mice/group) and ALT levels (B). C, The weight of mice was monitored before and after tumor challenge and throughout treatment (n = 5–15). D, The percentage of weight loss was calculated at day 7 after tumor challenge. E, The therapeutic index for the different treatments was calculated by dividing the percentage regression of mice observed per group and the average AST number and multiplying this by 100. Results represent pooled data from multiple experiments. Data for individual mice are shown along with mean ± SEM. Statistical significance was calculated using one-way ANOVA. *, P < 0.05; ***, P < 0.005; ****, P < 0.0005.

Article Snippet: The following antibodies for immunotherapy were purchased from BioXcell and used at the concentrations shown: α4–1BB (LOB12.3 at 350 μg per dose), αCD40 (FGK4.5 at 100 μg per dose), αCTLA-4 (9H10 at 100 μg per dose), αPD-1 (RMP1–14 at 250 μg per dose), and αOX-40 (OX-86 at 100 μg per dose).

Techniques:

Journal: Cancer research

Article Title: Mucosal HPV E6/E7 Peptide Vaccination in Combination with Immune Checkpoint Modulation Induces Regression of HPV + Oral Cancers

doi: 10.1158/0008-5472.CAN-18-0892

Figure Lengend Snippet: Combination of α4–1BB and αCTLA-4 provides tumor protection by increasing the ratios of CD8+ T cells to immunosuppressive populations. Mice bearing subcutaneous mEER tumors were either untreated or treated with intranasal HPV peptide vaccine, α4–1BB, or αCTLA-4 individually or in combinations as described in Fig. 1 legend. Tumors were harvested at day 15 and the isolated leukocytes were characterized by flow cytometry after staining for different surface and intracellular markers. The figure shows percentages of CD8+T cells (A), frequencies of CD8+ T cells as number of cells per tumor area (B), percentages of CD8+ T cells expressing GrzB and KLRG1 (C), frequencies of CD8+GrzB+KLRG1+ cells (D), percentages of CD4+FoxP3+ Tregs (E), frequencies of CD4+FoxP3+ Tregs (F), CD8+ T cells to Treg ratio (G), percentages of CD11b+Gr1+ MDSCs (H), frequencies of CD11b+Gr1+ MDSCs (I), and the CD8+ T cells to MDSC ratio (J). Data are represented as means ± SEM. Results represent pooled data from multiple experiments (n = 2–11 mice/group). Statistical significance was calculated using one-way ANOVA. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.00005.

Article Snippet: The following antibodies for immunotherapy were purchased from BioXcell and used at the concentrations shown: α4–1BB (LOB12.3 at 350 μg per dose), αCD40 (FGK4.5 at 100 μg per dose), αCTLA-4 (9H10 at 100 μg per dose), αPD-1 (RMP1–14 at 250 μg per dose), and αOX-40 (OX-86 at 100 μg per dose).

Techniques: Isolation, Flow Cytometry, Staining, Expressing

Journal: Cancer research

Article Title: Mucosal HPV E6/E7 Peptide Vaccination in Combination with Immune Checkpoint Modulation Induces Regression of HPV + Oral Cancers

doi: 10.1158/0008-5472.CAN-18-0892

Figure Lengend Snippet: Combination of HPV peptide vaccine, α4–1BB and αCTLA-4 induces regression of HPV+ tongue tumors. Mice were challenged with mEER tumor cells (40,000) in the tongue and treated with the different vaccine and immunotherapy combinations starting on day 5 as described in Fig. 1. A representative MRI (T2-weighted sagittal image) of mouse tongue (A) and tumor volume measured by MRI (B) at day 19 after tumor implantation are shown (data shown are means ± SEM; n = 5–12 mice/group). Survival curves of mice bearing tongue tumors treated with different immunotherapy combinations are shown (C; n = 10 – 23 mice/group). Statistical significance was calculated using one-way ANOVA. *, P < 0.05. Results represent pooled data from multiple experiments.

Article Snippet: The following antibodies for immunotherapy were purchased from BioXcell and used at the concentrations shown: α4–1BB (LOB12.3 at 350 μg per dose), αCD40 (FGK4.5 at 100 μg per dose), αCTLA-4 (9H10 at 100 μg per dose), αPD-1 (RMP1–14 at 250 μg per dose), and αOX-40 (OX-86 at 100 μg per dose).

Techniques: Tumor Implantation