ox86 agonist antibody  (Bio X Cell)

Journal: Neuro-Oncology Advances

Article Title: Enhanced glioblastoma immunotherapy via SMAC mimetic dose escalation and TGFβ blockade

doi: 10.1093/noajnl/vdaf253

Figure Lengend Snippet: High dose SMC remodels the GBM microenvironment . A) Experimental outline for IHC and flow cytometric analysis of IC GL261 tumors following high dose LCL161 treatments. Mice bearing IC GL261 tumors were treated orally 2×/week or 5×/week for two weeks with 100mg/kg LCL161. B) At day 18 post-implant, brains were processed for immunohistochemical staining of IBA1, TMEM119, and Arginase-1 (ARG1). Scale bar: 500 µm. C–G) Total IBA1 + cells (C) and TMEM119 subsets (D–G) within identified tumor margins were quantified and plotted as a percentage of total DAPI detections or as a fraction of total IBA1 + cells. H) Mice bearing IC GL261 tumors were stained for CD3, CD8, PD-1, TIM3 and OX40 expression. I–L) Total CD3 + cell counts, CD8 + subsets, and expression of TIM3 were quantified and plotted as either percent of total DAPI detections within tumor margins or as a proportion of total CD3+ events. N = 2 vehicle, N = 4 2d/wk, N = 5 5d/wk LCL161 treated animals per group *P < 0.05; ***P < 0.001; via one-way ANOVA using Tukey’s HSD multiple comparison test.

Article Snippet: Anti-mouse OX40 , 1:50, AR pH6 , Cell Signaling , 61637S.

Techniques: Immunohistochemical staining, Staining, Expressing, Comparison

Journal: Neuro-Oncology Advances

Article Title: Enhanced glioblastoma immunotherapy via SMAC mimetic dose escalation and TGFβ blockade

doi: 10.1093/noajnl/vdaf253

Figure Lengend Snippet: SMCs upregulate T-cell activation markers in peripheral lymphoid organs but not in brain tumors . A) Flow cytometric analysis of CD3 (BV786), CD4 (AF700) and CD8 (FITC) in spleen from mice bearing GL261 tumors and treated with vehicle or 100mg/kg LCL161 2d/week. B) Quantification of live cells CD3 + in spleen and lymph nodes across indicated treatment groups. N = 2 for unimplanted, N = 5 per remaining treatment groups. *P < 0.05; **P < 0.01 via two-way ANOVA using Tukey’s HSD multiple comparison test. C) Proportion of CD3 + CD4 + cells in indicated organ expressing CD25 receiving indicated treatments. D, E) MFI of OX40 (D) and CD69 (E) on CD3 + CD4 + and CD3 + CD8 + cells in indicated organ receiving indicated treatment. F) PD-1 expression of splenic CD8+ T-cells from mice with or without IC GL261 tumors, quantified in (G). H) MFI of LAG3 expression in CD3 + cells in spleen. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 via two-way ANOVA using Tukey’s HSD multiple comparison test. I–L) Flow cytometric analysis of CD3 positivity and subsets in resected GL261 tumors following treatment with vehicle or 100mg/kg LCL161 2d/wk. J) Quantification of proportion of live cells CD3 + . L) Ratio of CD4: CD8 cells among CD3 + populations.

Article Snippet: Anti-mouse OX40 , 1:50, AR pH6 , Cell Signaling , 61637S.

Techniques: Activation Assay, Comparison, Expressing

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Immunohistochemical and flow cytometric analysis of gastric cancer tissue. (A) H&E histochemical staining of tumor-free area (left), tumor margin (middle), and gastric tumor site (right). (B) Representative IHC images for CD3 + , CD4 + , CD8 + , FOXP3, OX40, and OX40L in tumor-free area (left column), tumor margin (middle column), and gastric tumor site (right column) sections. (C) Immunofluorescence staining for the co-expression of CD3 + and OX40 in gastric cancer with anti-CD3 and anti-OX40 antibodies and DAPI for nuclear staining. (D) Flow cytometry analysis of OX40 expression in CD3 cells obtained from PBMC of gastric cancer patients (upper panel) and OX40 expression on CD3 on TILs in gastric cancer patients (low panel). Statistical analysis of all data were performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; ***, P<0.001. ANOVA, analysis of variance; DAPI, 4',6-diamidino-2-phenylindole; H&E, hematoxylin & eosin; IHC, immunohistochemistry; OX40L, OX40 ligand; PBMC, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Expressing, Flow Cytometry, Standard Deviation, Immunohistochemistry

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: OX40L/IL-2 stimulation of PBMCs and TILs. (A) Flow cytometry analyses of T cell expression upon stimulation with different concentration of OX40L/IL-2. (B) Flow cytometry analyses of PBMCs and TILs upon stimulation with IL-2, OX40L, and OX40L/IL-2. (C) Expression at mRNA and secreted protein level of TNF-α, IFN-γ, Perforin, and Granzyme B after OX40L/IL-2 stimulation in PBMCs. (D) Expression at mRNA and secreted protein level of TNF-α, IFN-γ, Perforin, and Granzyme B after OX40L/IL-2 stimulation in TILs. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01; ***, P<0.001. ANOVA, analysis of variance; IFN-γ, interferon-gamma; IL-2, interleukin-2; OX40L, OX40 ligand; PBMCs, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Flow Cytometry, Expressing, Concentration Assay, Standard Deviation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Apoptotic effect of OX40L and IL-2 on co-culture of PBMCs or TILs from gastric cancer patients with gastric cancer primary cells. (A) H&E staining and immunohistochemistry labeling with a cytokeratin antibodies of primary gastric cancer cells. (B) Flow cytometry analyses of apoptosis in PBMCs co-cultured with primary gastric cancer cells alone, and after stimulation with either IL-2, or OX40L, or OX40L/IL-2. (C) Flow cytometry analyses of apoptosis in TILs co-cultured with primary gastric cancer cells alone and after stimulation with either IL-2, or OX40L, or OX40L/IL-2. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01. ANOVA, analysis of variance; H&E, hematoxylin & eosin; IL-2, interleukin-2; OX40L, OX40 ligand; PBMCs, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Co-Culture Assay, Staining, Immunohistochemistry, Labeling, Flow Cytometry, Cell Culture, Standard Deviation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Apoptotic effects of infectious adenovirus pAd-IL-2-OX40L on co-cultured gastric cancer patients TILs and gastric cancer primary cells. (A) Schematic presentation of adenoviral vector construction expressing OX40L/IL-2 and transfection of HEK293K cells with subsequent production of infectious AdV vector particles. (B) Flow cytometry analyses of gastric cancer primary cells, gastric cancer primary cells infected with pAd-IL-2-OX40L, gastric cancer primary cells and TILs, and co-culture of gastric cancer primary cells infected with pAd-IL-2-OX40L and TILs. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01. ANOVA, analysis of variance; IL-2, interleukin-2; OX40L, OX40 ligand; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Cell Culture, Plasmid Preparation, Expressing, Transfection, Flow Cytometry, Infection, Co-Culture Assay, Standard Deviation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Immunohistochemical and flow cytometric analysis of gastric cancer tissue. (A) H&E histochemical staining of tumor-free area (left), tumor margin (middle), and gastric tumor site (right). (B) Representative IHC images for CD3 + , CD4 + , CD8 + , FOXP3, OX40, and OX40L in tumor-free area (left column), tumor margin (middle column), and gastric tumor site (right column) sections. (C) Immunofluorescence staining for the co-expression of CD3 + and OX40 in gastric cancer with anti-CD3 and anti-OX40 antibodies and DAPI for nuclear staining. (D) Flow cytometry analysis of OX40 expression in CD3 cells obtained from PBMC of gastric cancer patients (upper panel) and OX40 expression on CD3 on TILs in gastric cancer patients (low panel). Statistical analysis of all data were performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; ***, P<0.001. ANOVA, analysis of variance; DAPI, 4',6-diamidino-2-phenylindole; H&E, hematoxylin & eosin; IHC, immunohistochemistry; OX40L, OX40 ligand; PBMC, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Expressing, Flow Cytometry, Standard Deviation, Immunohistochemistry

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: OX40L/IL-2 stimulation of PBMCs and TILs. (A) Flow cytometry analyses of T cell expression upon stimulation with different concentration of OX40L/IL-2. (B) Flow cytometry analyses of PBMCs and TILs upon stimulation with IL-2, OX40L, and OX40L/IL-2. (C) Expression at mRNA and secreted protein level of TNF-α, IFN-γ, Perforin, and Granzyme B after OX40L/IL-2 stimulation in PBMCs. (D) Expression at mRNA and secreted protein level of TNF-α, IFN-γ, Perforin, and Granzyme B after OX40L/IL-2 stimulation in TILs. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01; ***, P<0.001. ANOVA, analysis of variance; IFN-γ, interferon-gamma; IL-2, interleukin-2; OX40L, OX40 ligand; PBMCs, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Flow Cytometry, Expressing, Concentration Assay, Standard Deviation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Apoptotic effect of OX40L and IL-2 on co-culture of PBMCs or TILs from gastric cancer patients with gastric cancer primary cells. (A) H&E staining and immunohistochemistry labeling with a cytokeratin antibodies of primary gastric cancer cells. (B) Flow cytometry analyses of apoptosis in PBMCs co-cultured with primary gastric cancer cells alone, and after stimulation with either IL-2, or OX40L, or OX40L/IL-2. (C) Flow cytometry analyses of apoptosis in TILs co-cultured with primary gastric cancer cells alone and after stimulation with either IL-2, or OX40L, or OX40L/IL-2. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01. ANOVA, analysis of variance; H&E, hematoxylin & eosin; IL-2, interleukin-2; OX40L, OX40 ligand; PBMCs, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Co-Culture Assay, Staining, Immunohistochemistry, Labeling, Flow Cytometry, Cell Culture, Standard Deviation

Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Apoptotic effects of infectious adenovirus pAd-IL-2-OX40L on co-cultured gastric cancer patients TILs and gastric cancer primary cells. (A) Schematic presentation of adenoviral vector construction expressing OX40L/IL-2 and transfection of HEK293K cells with subsequent production of infectious AdV vector particles. (B) Flow cytometry analyses of gastric cancer primary cells, gastric cancer primary cells infected with pAd-IL-2-OX40L, gastric cancer primary cells and TILs, and co-culture of gastric cancer primary cells infected with pAd-IL-2-OX40L and TILs. Statistical analysis of all data was performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; **, P<0.01. ANOVA, analysis of variance; IL-2, interleukin-2; OX40L, OX40 ligand; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Cell Culture, Plasmid Preparation, Expressing, Transfection, Flow Cytometry, Infection, Co-Culture Assay, Standard Deviation