Journal: Frontiers in Immunology
Article Title: Antagonistic Antibody Targeting TNFR2 Inhibits Regulatory T Cell Function to Promote Anti-Tumor Activity
Figure Lengend Snippet: AN3025 inhibits TNFα-TNFR2 mediated cell death of hTNFR2 overexpressing Jurkat cell and enhances T effector cell function in Tregs/Teff co-culture assay. (A) Expression of human TNFR2 on wildtype Jurkat cells. (B) Viability of wildtype Jurkat cells after stimulation with titrated human TNFα. (C) Expression of human TNFR2 on established hTNFR2 overexpressing Jurkat single cell clone. (D) Viability of hTNFR2 overexpressing Jurkat cells after stimulation with titrated human TNFα. (E) Viability of hTNFR2 overexpressing Jurkat cells after stimulation with constant 0.5ng/mL human TNFα in the presence of titrated AN3025 or titrated human IgG 1 , κ. (F) Competitive ELISA binding assay of AN3025 with biotinylated human TNFα to human TNFR2 on plate. (G, H) Human iTregs were co-cultured with CFSE-labeled autologous CD4 + Teff cells for 4 days. (G) CFSE lo T effector cell percentage was quantified by flow cytometry. (H) IFNγ in the supernatants was detected by ELISA assay. Values were expressed as Mean ± SEM. **P < 0.01; ***P < 0.001.
Article Snippet: Human TNFR2 (Acro Biosystem, TN2-H5227), human TNFR1 (Acro Biosystem, TN1-H5222), cynomolgus TNFR2 (Sino Biological,90102-C08H), mouse TNFR2 (R&D systems, 426-R2-050/CF) or rat TNFR2 (R&D systems, 8348-R2-050) was coated on high binding polystyrene flat bottom micro-titer plates (Thermo Scientific, 3455).
Techniques: Cell Function Assay, Co-culture Assay, Expressing, Competitive ELISA, Binding Assay, Cell Culture, Labeling, Flow Cytometry, Enzyme-linked Immunosorbent Assay