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>The mouse model and efficacy evaluation of immunological checkpoint

Gene description

OX40, also known as Tnfrsf4 (Tumor necrosis factor receptor super family, member 4), is mainly expressed on the surface of activated CD4+ and CD8+ T cells, and its binding with the OX40 ligand can stimulate CD8+ T cell activation. The coactivation of OX40/OX40L enhances T cell function, including cytokine production, proliferation and T cell survival. An OX40 agonist can reduce Regulatory T cells (Tregs) and improve anti-tumor activity.

mRNA expression analysis

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Fig 1. RT-PCR analysis of OX40 gene. The hOX40, but not mOX40, mRNA was detected in splenocytes of the homozygous B-hOX40 mice.

Protein expression analysis

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Fig 2. Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hOX40 mice were analyzed by flow cytometry. Mouse OX40+ T cells were detected in the WT mice, while human OX40+ T cells were detected in the homozygous B-hOX40 mice.

Human OX40 mAb efficacy evaluation (MC38 cell line)

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Fig 3. Murine colon cancer cells (MC38) were subcutaneously implanted into heterozygous B-hOX40 mice. Mice were divided into control and experimental groups (n=5) when the tumor size was about 100 mm3. Anti-hOX40 antibody significantly inhibited tumor growth in the heterozygous B-hOX40 mice at 3 different doses, suggesting that the B-hOX40 mouse model is an effective tool for in vivo hOX40 antibody efficacy studies. (A) Tumor average volume ± SEM, (B) Mice average weight ± SEM.

OX40 Ab efficacy evaluation (MC38 cell line)

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Fig 4. Murine colon cancer cells (MC38) were subcutaneously implanted into homozygous B-hOX40 mice. Mice were divided into control and experimental groups (n=5) when the tumor size was about 100 mm3. The aAnti-hOX40 antibody significantly inhibited tumor growth in the homozygous B-hOX40 mice, suggesting that the B-hOX40 mouse model is an effective tool for in vivo hOX40 antibody efficacy studies. (A) Tumor average volume ± SEM, (B) Mice average weight ± SEM.

Human OX40 (MOXR0916 Analog) mAb efficacy evaluation (MC38 cell line)

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Fig 5. Human OX40 antibody (MOXR0916 analog) efficacy experiment was performed with B-hOX40 mice.

Colon cancer cells were subcutaniously implanted into the homozygous B-hOX40 mice. The mice were divided into control and treatment groups as the tumor volume was 150±50mm3 (n=5). The result indicated that B-hOX40 mouse model was an effective tool for in vivo hOX40 antibody efficacy study. The average±SEM of tumor sizes are shown in the figure.


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