agonist anti ox40  (Bio X Cell)

Journal: Journal of Translational Medicine

Article Title: Enhanced systemic tumor suppression by in situ vaccine combining radiation and OX40 agonist with CpG therapy

doi: 10.1186/s12967-023-04504-w

Figure Lengend Snippet: CpG and radiotherapy upregulated the expression of OX40 on tumor infiltrating CD4+ T cells in the tumor microenvironment. A The correlation between OX40 expression and TLR9 or TLR9 downstream elements in SKCM was analyzed by an online tool GEPIA 2. B , C 48 h after treated with PBS, CpG intratumoral injection or 10 Gy irradiation followed by CpG intratumoral injection, tumors were excised and OX40 expression on the CD45+CD4+ T cell subset was analyzed by flow cytometry (OX40+ subset gate on CD45+CD4+ cells). Data are represented as mean ± s.e.m. n = 4. Student’s t test was used for statistical analysis. * p < 0.05

Article Snippet: Anti-OX40 agonistic antibody (αOX40, Clone OX-86) was provided by GenScript USA Inc. Murine anti-PD-1 antibody (G4C2) was provided by Shanghai Junshi Biosciences Co.,Ltd (Suzhou, China).Monoclonal antibodies (mAbs) used for flow cytometry were listed as flow: FITC anti-mouse CD45 (Biolegend, USA), PE/DazzleTM anti-mouse CD3 (Biolegend, USA), FITC anti-mouse CD8 (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse CD4 (Biolegend, USA), FITC anti-mouse CD11c (Biolegend, USA), PE anti-mouse CD86 (Biolegend, USA), PE/Cyanine7 anti-mouse CD80 (Biolegend, USA), PE anti-mouse CD44 (Biolegend, USA), PE/Cyanine7 anti-mouse CD62L (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse PD-1 (Biolegend, USA), PE/Cyanine7 anti-mouse OX40 (Biolegend, USA), FITC anti-mouse CD11b (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse F4/80 (Biolegend, USA), APC anti-mouse CD206 (Biolegend, USA), Mouse Regulatory T cell staining kit (eBioscience, USA).

Techniques: Expressing, Injection, Irradiation, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Enhanced systemic tumor suppression by in situ vaccine combining radiation and OX40 agonist with CpG therapy

doi: 10.1186/s12967-023-04504-w

Figure Lengend Snippet: Systemic immune response induced by triple combining in situ vaccine. A , B Proportion of effector memory T cells (TEMs, CD44+CD62L– gate on CD3+CD8+ cells) in A abscopal tumors and B spleens of C57BL/6 mice in each group a week after the last administration. Data are represented as mean ± s.e.m. n = 5–6. Student’s t test was used for statistical analysis. * p < 0.05, ** p < 0.01. C Spleen cells of mice in control group and RT + CpG + αOX40 group were incubated with CFSE labeled (indicating live cells) B16F10 melanoma cells at effector-to-target ratio (E:T) of 5:1, 10:1, 20:1 and 30:1. PI was added after 5 h-incubation and the percentage of dead cells was analyzed by flow cytometry. Data are represented as mean ± s.e.m. n = 4. Student’s t test was used for statistical analysis. * p < 0.05. D Representative flow cytometry plots indicating proportions of dead cells of B16F10 melanoma cells (CFSE + PI + gate on CFSE + cells). E , F Proportion of E PD-1+ CTLs (CD8+PD-1+ gate on CD3+ cells) and F regulatory T cells (Tregs, CD25+FoxP3+ gate on CD3+CD4+ cells) in abscopal tumors of C57BL/6 mice in each group a week after the last administration. Data are represented as mean ± s.e.m. n = 3–5. Student’s t test or corrected Student’s t test was used for statistical analysis. * p < 0.05

Article Snippet: Anti-OX40 agonistic antibody (αOX40, Clone OX-86) was provided by GenScript USA Inc. Murine anti-PD-1 antibody (G4C2) was provided by Shanghai Junshi Biosciences Co.,Ltd (Suzhou, China).Monoclonal antibodies (mAbs) used for flow cytometry were listed as flow: FITC anti-mouse CD45 (Biolegend, USA), PE/DazzleTM anti-mouse CD3 (Biolegend, USA), FITC anti-mouse CD8 (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse CD4 (Biolegend, USA), FITC anti-mouse CD11c (Biolegend, USA), PE anti-mouse CD86 (Biolegend, USA), PE/Cyanine7 anti-mouse CD80 (Biolegend, USA), PE anti-mouse CD44 (Biolegend, USA), PE/Cyanine7 anti-mouse CD62L (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse PD-1 (Biolegend, USA), PE/Cyanine7 anti-mouse OX40 (Biolegend, USA), FITC anti-mouse CD11b (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse F4/80 (Biolegend, USA), APC anti-mouse CD206 (Biolegend, USA), Mouse Regulatory T cell staining kit (eBioscience, USA).

Techniques: In Situ, Control, Incubation, Labeling, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Enhanced systemic tumor suppression by in situ vaccine combining radiation and OX40 agonist with CpG therapy

doi: 10.1186/s12967-023-04504-w

Figure Lengend Snippet: RNA-seq analysis of control tumors and tumors receiving in situ triple vaccine. A Heat map revealing in situ vaccine-induced changes in the gene-expression profile of the tumor microenvironment. n = 3 for both control and RT + CpG + αOX40 group. Color bars indicate normalized expression, the color from blue to red indicates that the gene expression from low to high. B GO enrichment analysis of up- and downregulated genes. Gene ratio (shown in green) and − log10(Q value) of all GO terms are shown. C KEGG enrichment analysis of up- and downregulated genes. Gene ratio (shown in green) and − log10(Q value) of all KEGG signaling pathway are shown. D Representative GSEA plot showing down-regulated and up-regulated enriched gene sets of all detected genes in tumors receiving in situ triple vaccine

Article Snippet: Anti-OX40 agonistic antibody (αOX40, Clone OX-86) was provided by GenScript USA Inc. Murine anti-PD-1 antibody (G4C2) was provided by Shanghai Junshi Biosciences Co.,Ltd (Suzhou, China).Monoclonal antibodies (mAbs) used for flow cytometry were listed as flow: FITC anti-mouse CD45 (Biolegend, USA), PE/DazzleTM anti-mouse CD3 (Biolegend, USA), FITC anti-mouse CD8 (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse CD4 (Biolegend, USA), FITC anti-mouse CD11c (Biolegend, USA), PE anti-mouse CD86 (Biolegend, USA), PE/Cyanine7 anti-mouse CD80 (Biolegend, USA), PE anti-mouse CD44 (Biolegend, USA), PE/Cyanine7 anti-mouse CD62L (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse PD-1 (Biolegend, USA), PE/Cyanine7 anti-mouse OX40 (Biolegend, USA), FITC anti-mouse CD11b (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse F4/80 (Biolegend, USA), APC anti-mouse CD206 (Biolegend, USA), Mouse Regulatory T cell staining kit (eBioscience, USA).

Techniques: RNA Sequencing, Control, In Situ, Gene Expression, Expressing