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>The mouse model and efficacy evaluation of immunological checkpoint

Gene description

TNFRSF9 (TNF receptor super family member 9), also called CD137 or 4-1BB, is mainly expressed on the surface of activated T, NK, and mononuclear cells. CD137 is activated by its ligand CD137L or an activating CD137 monoclonal antibody, and stimulates T cell activation, proliferation and cytokine production. This co-stimulatory signal can also multiply antigen presenting cells and result in cell factor secretion. Experiments show that CD137 co-stimulation regulates T cell and antigen presenting cell function for antitumor immunity, providing new targets for tumor immunological therapies.

mRNA expression analysis

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Fig 1. RT-PCR analysis of 4-1BB gene.

Theh4-1BB, but not m4-1BB, mRNA  was detected in splenocytesof the homozygous B-h4-1BB mice.

Protein expression analysis

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Fig 2. Splenocytes from both wild type (WT) C57BL/6 and homozygous B-h4-1BB mice were analyzed by flow cytometry. Mouse 4-1BB+ T cells were detected in the WT mice, while human 4-1BB+ T cells were detected in the homozygous B-h4-1BB mice.

Human 4-1BB Ab (Urelumab) mAb efficacy evaluation (MC38 cell line) 

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Fig3. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-h4-1BB mice. Mice were divided into control and treatment groups (n=6), when the tumor size was approximately 150±50 mm3. Three doses Urelumab showed substantially inhibitory effects in the homozygous B-h4-1BB mice, suggesting that B-h4-1BB mouse model is a useful tool for in vivo h4-1BB antibody efficacy study. The average ± SEM of tumor sizes are shown in the figure.

Humna 4-1BB (Urelumab Analog) mAb toxicity evaluation

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Fig4.The blood routine examination was performed with anti-human 4-1BB antibody on B-h4-1BB mice at the termination of efficacy evaluation experiment.


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Humna 4-1BB (Urelumab Analog) mAb toxicity evaluation


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Fig5.The blood biochenmical index was examined with anti-human 4-1BB antibody on B-h4-1BB mice at the termination of efficacy evaluation experiment.

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Human 4-1BB (Utomilumab Analog) mAb Efficacy Evaluation (MC38 cell line)

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Fig6. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-h4-1BB mice. Mice were divided into control and treatment groups (n=6), when the tumor size was approximately 150±50 mm3. The result indicated that substantially inhibitory effects in the homozygous B-h4-1BB mice, suggesting that B-h4-1BB mouse model is a useful tool for in vivo h4-1BB antibody efficacy study. The average ± SEM of tumor sizes are shown in the figure.

Human 4-1BB (Utomilumab Analog) mAb toxicity Evaluation

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Fig7.The blood biochenmical index was examined with anti-human 4-1BB antibody on B-h4-1BB mice at the termination of efficacy evaluation experiment.

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