m1650 mouse anti tnfr2  (Hycult Biotech)

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: Characterizations of the different PLGA nanoparticles

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Zeta Potential Analyzer, Encapsulation

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: Characterization of PLGA-ADR, ISO-PLGA-ADR, and TNFR2-PLGA-ADR nanoparticles. (A to C) Size distribution and scanning electron microscopy image of PLGA-ADR (A), ISO-PLGA-ADR (B), and TNFR2-PLGA-ADR (C) nanoparticles. Scale bar, 200 nm. (D) In vitro release curves of ADR from ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles in PBS at pH 7.4 ( n = 3). (E) Particle size of ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles at 4°C for different periods determined by DLS ( n = 3). (F) Size of ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles in PBS (pH 7.4) containing 10% FBS at 37°C for different periods determined by DLS ( n = 3).

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Electron Microscopy, In Vitro

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: Binding of ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles with CT26 and MC38 colon cancer cells. CT26 and MC38 cells were incubated at 37°C for 4 h with ISO-PLGA-ADR, TNFR2-PLGA-ADR, or TNFR2-PLGA-ADR plus anti-TNFR2 antibody (25 μg/ml) pretreated for 1 h (equivalent ADR concentration of 1 μg/ml). The cellular binding was determined by the expression of ADR by using flow cytometry. Representative FACS plots (A) and summary of mean fluorescence intensity [MFI; (B)] are shown. (C) CT26 and MC38 cells were incubated at 37°C for 4 h with ISO-PLGA-ADR, TNFR2-PLGA-ADR, or TNFR2-PLGA-ADR plus anti-TNFR2 antibody (25 μg/ml) pretreated for 1 h, followed by staining with Hoechst 33258 (blue). The intracellular and nuclear accumulation of ADR (red) was determined and photographed with a confocal microscope. Data [means ± SEM (standard error of the mean), n = 3] shown are representatives of 3 separate experiments with similar results. By comparison with the indicated group, ** P < 0.01, *** P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Binding Assay, Incubation, Concentration Assay, Expressing, Flow Cytometry, Fluorescence, Staining, Microscopy, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The cytotoxic effects of ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles on CT26 and MC38 tumor cells. (A and B) Cell viability of CT26 (A) and MC38 (B) cells was examined by MTT assay after treatment with PLGA-PEG, ISO-PLGA-ADR, and TNFR2-PLGA-ADR nanoparticles at different concentrations for 24 h. (C and D) Apoptosis of CT26 and MC38 cells analyzed by FACS after treatments with ISO-PLGA-ADR and TNFR2-PLGA-ADR nanoparticles (ADR: 1 μg/ml) for 24 h. The typical FACS plots (C) and summary of the proportion of apoptotic cells in each group (D). Data (means ± SEM, n = 5) shown are representatives of 3 separate experiments with similar results. By comparison with the ISO-PLGA-ADR group, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: MTT Assay, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The distribution of ISO-PLGA-DiR and TNFR2-PLGA-DiR in mouse colon cancer. (A) Distribution of ISO-PLGA-DiR and TNFR2-PLGA-DiR. CT26 tumor-bearing BALB/c mice and MC38 tumor-bearing C57BL/6 mice were intravenously injected with fluorescent dye-labeled ISO-PLGA-DiR and TNFR2-PLGA-DiR as described in Materials and Methods when the volume of tumor reached 300 to 500 mm 3 , respectively. The distribution of ISO-PLGA-DiR and TNFR2-PLGA-DiR was examined and recorded with an in vivo imaging system for 1 to 48 h after injection. (B and C) Distribution of ISO-PLGA-DiR and TNFR2-PLGA-DiR in different organs. The mice were sacrificed 48 h after injection. The heart, liver, spleen, lungs, kidneys, and tumor were harvested, and the distribution of ISO-PLGA-DiR and TNFR2-PLGA-DiR was recorded using an in vivo imaging system (B). The average fluorescent intensity was quantified using the Maestro software (C). Data (mean ± SEM, n = 3) shown are representatives of 3 separate experiments with similar results. By comparison with ISO-PLGA-DiR, *** P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Injection, Labeling, In Vivo Imaging, Software, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The colocalization of ISO-PLGA-ADR and TNFR2-PLGA-ADR with tumor-infiltrating T regs . Foxp3 EGFP/DTR C57BL/6-Tg mice were inoculated in the right flank with MC38 cells (500,000 cells in 0.1 ml of PBS). When the size of tumor reached ~400 mm 3 , MC38 tumor-bearing Foxp3 EGFP/DTR C57BL/6-Tg mice were intravenously injected with ADR, ISO-PLGA-ADR, or TNFR2-PLGA-ADR. After 24 h, the frozen section of different tumor tissue was stained with Hoechst 33258. (A) Levels of ADR (red) and EGFP (green, T regs ) were measured using a confocal microscope (Zeiss 810). (B) Semiquantitative analysis for colocalization. Scale bar, 50 μm. Data shown are representatives of 3 separate experiments with similar results.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Injection, Staining, Microscopy

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: TNFR2-PLGA-ADR nanoparticles inhibit growth of mouse colon cancers. BALB/c mice were inoculated with CT26 cells (200,000 cells in 0.1 ml of PBS), and C57BL/6 mice were inoculated with MC38 cells (500,000 cells in 0.1 ml of PBS). On day 6 after tumor inoculation, CT26 tumor-bearing BALB/c mice and MC38 tumor-bearing C57BL/6 mice were randomly divided into 4 groups and intravenously injected with PBS, free ADR, ISO-PLGA-ADR, or TNFR2-PLGA-ADR every 3 d for 5 times, respectively. (A) The tumor size was monitored every other day. Tumor weight (B) was measured at the end of the experiment. Cell apoptosis (C) in the tumor tissue was evaluated by the TUNEL assay. Scale bars, 100 nm for CT26 tumor and 75 nm for MC38 tumor. (D) Semiquantitative analysis for cell apoptosis. The experiment was repeated for 3 times with similar results. Data shown are mean ± SEM ( n = 6). By comparison with the PBS group, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. By comparison with the indicated group, # P < 0.05, ## P < 0.01, ### P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Injection, TUNEL Assay, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The in vivo effects of TNFR2-PLGA-ADR nanoparticles on T regs . CT26 tumor-bearing BALB/c mice and MC38 tumor-bearing C57BL/6 mice were treated as described in Fig. . (A to C) The proportion of T regs in CD4 + cells and the expression of TNFR2 by T regs were analyzed by FACS, gating for Foxp3 + cells. Representative FACS plots (A) and summary of proportion of T regs (B). (C) Abundance of TNFR2 on the surface of T regs in the tumor. The data (mean ± SEM, n = 6) shown in (B) and (C) are representative of 3 separate experiments with similar results. By comparison with the PBS group, * P < 0.05, ** P < 0.01, *** P < 0.001. By comparison with the indicated group, # P < 0.05, ### P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: In Vivo, Expressing, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The in vivo effects of TNFR2-PLGA-ADR nanoparticles on tumor-infiltrating IFNγ + CD8 + CTLs. CT26 tumor-bearing BALB/c mice and MC38 tumor-bearing C57BL/6 mice were treated as described in Fig. . Single cells from tumor were restimulated in vitro, and intracellular IFNγ expression by CD8 + T cells was analyzed by FACS. Representative FACS plots (A) and summary (B). Bars (mean ± SEM, n = 6) shown are representative of 3 separate experiments with similar results. By comparison with the PBS group, * P < 0.05, ** P < 0.01, *** P < 0.001. By comparison with the indicated group, # P < 0.05, ### P < 0.001.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: In Vivo, In Vitro, Expressing, Comparison

Journal: Research

Article Title: Anti-TNFR2 Antibody-Conjugated PLGA Nanoparticles for Targeted Delivery of Adriamycin in Mouse Colon Cancer

doi: 10.34133/research.0444

Figure Lengend Snippet: The systemic toxicity of TNFR2-PLGA-ADR nanoparticles in tumor-bearing mice. CT26 tumor-bearing BALB/c mice and MC38 tumor-bearing C57BL/6 mice were treated as described in Fig. . (A) Body weight of mice. (B and C) Forty-eight hours after the last injection, serum levels of AST (B) and ALT (C) were measured by using a commercial assay kit. (D) Histologic results of representative liver tissues from mice treated with PBS, ADR, ISO-PLGA-ADR, or TNFR2-PLGA-ADR (200×). Scale bar, 100 μm. Data (mean ± SEM, n = 6) shown are representative of 3 separate experiments with similar results. By comparison with the PBS group, * P < 0.05, ** P < 0.01. By comparison with the indicated group, # P < 0.05, ## P < 0.01.

Article Snippet: InVivoMAb anti-mouse TNFR2 (CD120b, BE0247) and InVivoMAb polyclonal Armenian hamster IgG (isotype control, BE0091) were purchased from BioXCell. mPEG2k-PLGA5k (PLGA, copolymer ratio 50:50) and NHS-PEG2k-PLGA5k (PLGA, copolymer ratio 50:50) were purchased from Shanghai Mao Kang Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Injection, Comparison

Journal: bioRxiv

Article Title: TNFR2 loss leads to decreased TOX expression in T cells without affecting TIM3 and improves responses to tumor and chronic LCMV

doi: 10.1101/2024.07.12.603311

Figure Lengend Snippet: (A and B) TNFR1 (A) and TNFR2 (B) expression on CD8 + T cells harvested from the tumor, spleen, or tumor-draining lymph nodes from tumor-bearing mice over time, across tumor types and locations. N = 10-35 per time point. ǂ represents p < 0.0001 between tumor and spleen and between tumor and tumor-draining lymph node. Ɵ represents p < 0.01 between tumor and tumor-draining lymph node and p < 0.0001 between tumor and spleen. (C and D) Correlation of TNFR2 expression on tumor-infiltrating CD8 + T cells with expression of canonical markers of exhaustion (PD1, SLAMF6, TIM3, and TOX) in subcutaneous E0771 (C) and intracranial CT2A (D). Tumors were harvested at similar timepoints as displayed in . A simple linear regression model was performed to determine significance and R values. (E) Expression of TNFR2 on CD8 + T cells in the spleen of mice infected with chronic LCMV at day 7 and 14 post-infection compared to naïve spleen. One-way ANOVA was performed. (F-H) Expression of PD1 (F), TIM3 (F), and TOX (H) on TNFR2- and TNFR2+ CD8 + T cells in the spleen of mice infected with chronic viral infection at peak viral titers (D14). Paired t-test was performed. (I-K) Single cell RNA sequencing was performed on CD3 + T cells sorted from intracranial CT2A at day 13 and day 19 post tumor implantation. Following QC, CD8 + T cells clustered into 6 populations (I). Trajectory analysis was performed on the clusters identified in I (J). Using the pathway identified in J, expression of Pdcd1 (gene encoding PD1), Tcf7 (gene encoding progenitor-associated transcription factor TCF1), Havcr2 (gene encoding TIM3), and Tnfrsf1b (gene encoding TNFR2) was determined in the clusters Tex_prog, Tex_int, and Tex_late. ns not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: To determine the role of CD8 + T cells in the tumor growth phenotype observed in TNFR2 KO mice, CD8 + T cells were depleted through intraperitoneal injections of 200µg anti-CD8a (BioXCell, clone 2.43) on days - 3, -1, and every 6 days post tumor implantation.

Techniques: Expressing, Infection, RNA Sequencing Assay, Tumor Implantation

Journal: bioRxiv

Article Title: TNFR2 loss leads to decreased TOX expression in T cells without affecting TIM3 and improves responses to tumor and chronic LCMV

doi: 10.1101/2024.07.12.603311

Figure Lengend Snippet: (A) Experimental design to assess cytokine production in antigen-experienced (PD1 + ) TNFR2 - and TNFR2 + CD8 + T cells. Each tumor was harvested at a time point that produced sufficient TNFR2 - and TNFR2 + CD8 + T cells to allow sorting of each population. (B) Representative flow plots of IFNγ + TNF + (polyfunctional), IL-2, and granzyme B production of PD1 + CD8 + T cells. (C-D) Assessment of the progenitor-like functions polyfunctionality (IFNγ + TNF + ) (C) and IL-2 expression (D) in TNFR2 - and TNFR2 + CD8 + T cells harvested from tumors at time points displayed in (A) following stimulation with PMA, Ionomycin, GolgiStop, and GolgiPlug for 4 hours at 37°C. Paired t-test was performed. (E-F) Assessment of the terminal-like functions IFNγ + single positive (TNF - ) (E) and granzyme B (F) in TNFR2 - and TNFR2 + CD8 + T cells harvested from tumors at time points displayed in (A) following stimulation with PMA, Ionomycin, GolgiStop, and GolgiPlug for 4 hours at 37°C. Paired t-test was performed. ns not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: To determine the role of CD8 + T cells in the tumor growth phenotype observed in TNFR2 KO mice, CD8 + T cells were depleted through intraperitoneal injections of 200µg anti-CD8a (BioXCell, clone 2.43) on days - 3, -1, and every 6 days post tumor implantation.

Techniques: Produced, Expressing

Journal: bioRxiv

Article Title: TNFR2 loss leads to decreased TOX expression in T cells without affecting TIM3 and improves responses to tumor and chronic LCMV

doi: 10.1101/2024.07.12.603311

Figure Lengend Snippet: (A) Frequency of progenitor (SLAMF6 + TIM3 - ) and terminally exhausted (SLAMF6 - TIM3 + ) CD8 + TILs from wild type (WT) and TNFR2 knock out (KO) in subcutaneous models of CT2A (D35, top), E0771 (D25, middle), and cLCMV (D14, bottom). Two-way repeated measures ANOVA was performed. (B) Representative flow plot of TOX expression in PD1 - (green) and PD1 + TIM3 + (purple) in CD8 + TILs (left) and a histogram comparing TOX expression in WT (gray) and TNFR2 KO (blue) in PD1 + TIM3 + CD8 + TILs (right) (C) Mean fluorescence intensity (MFI) of TOX in WT and TNFR2 KO TIM3 + CD8 + T cells harvested from tumor for sc E0771 (D25), ic CT2A (D21), sc CT2A (D35), sc Yummer (D28), ic Yummer (D25), and spleen for cLCMV (D14). Unpaired t-test was performed. (D-E) Experimental design to assess T cell intrinsic requirement of TNFR2 (D). CT2A-Trp2 was implanted subcutaneously into the flank of CD45.1 + CD45.2 + mice. After 21 days, Trp2-specific T cells generated from WT (CD45.1) and TNFR2 KO (CD45.2) mice were injected intravenously at a 1:1 ratio. Tumors were harvested on day 28 and transferred Trp2-specific WT (CD45.1) and TNFR2 KO (CD45.2) T cells were assessed for TOX expression (E). Paired t-test was performed. (F-I) Polyfunctionality (IFNγ + TNF + ) (F), IL-2 (G), Granzyme B (H), and IFNγ + single positive (I) was assessed on CD8 + T cells from splenocytes of WT and TNFR2 KO mice following chronic LCMV infection (D14) following stimulation with PMA, Ionomycin, GolgiStop, and GolgiPlug for 4 hours at 37°C. Unpaired t-test was performed. ns not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: To determine the role of CD8 + T cells in the tumor growth phenotype observed in TNFR2 KO mice, CD8 + T cells were depleted through intraperitoneal injections of 200µg anti-CD8a (BioXCell, clone 2.43) on days - 3, -1, and every 6 days post tumor implantation.

Techniques: Knock-Out, Expressing, Fluorescence, Generated, Injection, Infection

Journal: bioRxiv

Article Title: TNFR2 loss leads to decreased TOX expression in T cells without affecting TIM3 and improves responses to tumor and chronic LCMV

doi: 10.1101/2024.07.12.603311

Figure Lengend Snippet: (A) Experimental design to assess transcriptional program of WT and TNFR2 KO CD8 + TIM3 + T cells. CD8 + PD1 + SLAMF6 - TIM3 + splenocytes were sorted from WT and TNFR2 KO mice 14 days post infection with cLCMV. (B) PCA plot representing distribution of WT and TNFR2 KO CD8 + TIM3 + T cells based on gene expression profile. (C) MA-plot highlighting significantly downregulated (blue) and significantly upregulated genes in TNFR2 KO CD8 + TIM3 + T cells. (D) Gene module enrichment analysis was performed based on the mouse NanoString nCounter® Immune Exhaustion Panel (genes listed in supplementary table 2) . Pathways highlighted in blue were statistically downregulated using a corrected p-value <0.1. (E) Heatmaps displaying z-scores for exhausted-related genes. (F) Gene expression levels for Tox in WT and TNFR2 KO CD8 + TIM3 + T cells. Unpaired t-test was performed. (G-H) Heatmaps displaying z-scores for AP1-transcription factors and TOX regulators (G), and epigenetic modifiers (H) for each WT and TNFR2 KO CD8 + TIM3 + sample. * p < 0.05, ** p < 0.01

Article Snippet: To determine the role of CD8 + T cells in the tumor growth phenotype observed in TNFR2 KO mice, CD8 + T cells were depleted through intraperitoneal injections of 200µg anti-CD8a (BioXCell, clone 2.43) on days - 3, -1, and every 6 days post tumor implantation.

Techniques: Infection, Expressing

Journal: bioRxiv

Article Title: TNFR2 loss leads to decreased TOX expression in T cells without affecting TIM3 and improves responses to tumor and chronic LCMV

doi: 10.1101/2024.07.12.603311

Figure Lengend Snippet: (A-B) Tumor growth curves for WT and TNFR2 KO mice following subcutaneous implantation of E0771 (A) and CT2A (B). Tumors were measured every 3 days starting on day 9. Two-way repeated ANOVA was performed. (C) Experimental design to determine the role of CD8 T cell depletion on tumor control in TNFR2 KO mice. CD8 T cells were depleted with a depletion antibody day -3, day -1, and every 6 days post tumor implantation. CT2A was implanted subcutaneously on day 0. (D) Tumor growth curves for WT, TNFR2 KO, and CD8-depleted TNFR2 KO mice following subcutaneous implantation of CT2A. Tumors were measured every 3 days starting on day 9. Two-way repeated ANOVA was performed. (E-F) Tumor growth for WT mice dosed with anti-PD1, anti-TNFR2, or a combination of anti-PD1 and anti-TNFR2 following subcutaneous implantation of E0771 (E) or CT2A-Trp2 (F). Mice were injected with anti-PD1 (200ug), anti-TNFR2 (250ug), or both every 3 days starting on day 9. Tumors were measured every 3 days. Two-way repeated ANOVA was performed. (G-H) Time to humane endpoint for mice displayed in E and F following subcutaneous implantation of E0771 (G) and CT2A-Trp2 (H). Log-rank test was performed between each group for a total of 6 comparisons. A corrected p-value of less than 0.0083 (p < 0.05/6 comparisons) was used as a cutoff for significance. Mice were followed for 80 days. If no tumor was detected at day 80, the mouse was considered a long-term survivor. * p < 0.05, ** p < 0.01, *** p<0.001, **** p < 0.0001

Article Snippet: To determine the role of CD8 + T cells in the tumor growth phenotype observed in TNFR2 KO mice, CD8 + T cells were depleted through intraperitoneal injections of 200µg anti-CD8a (BioXCell, clone 2.43) on days - 3, -1, and every 6 days post tumor implantation.

Techniques: Control, Tumor Implantation, Injection