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>The mouse model and efficacy evaluation of immunological checkpoint

Gene description

TIM3 (T-cell immunoglobulin domain and mucindomain-3) is an activation-induced inhibitory molecule involved in immune tolerance and T-cell exhaustion in chronic viral infection and cancers. TIM3 maturation and cell surface expression is facilitated by forming a heterodimeric interaction with CD66a. Co-blockade of CD66a and TIM3 enhanced anti-tumor immune responses, and eliminated tumors in mouse colorectal cancer models.

mRNA expression analysis

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Fig 1. RT-PCR analysis of TIM3 gene.

The hTIM3, but not mTIM3, mRNA  was detected in splenocytesof the homozygous B-hTIM3 mice.

Protein expression analysis

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Fig 2. Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hTIM3 mice were analyzed by flow cytometry. Mouse TIM3+ cells were detected in non-T and non-B cells from the WT mice, while human TIM3+ cells were detected in non-T and non-B cells from the homozygous B-hTIM3 mice.

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Fig 3. Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hTIM3 mice were analyzed by flow cytometry. Mouse TIM3+ T cells were detected in the WT mice, while human TIM3+T cells were detected in the homozygous B-hTIM3 mice.

Human TIM3 mAb efficacy evaluation (MC38 cell line)

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Fig 4. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hTIM3 mice. Mice were divided into control and treatment groups (n = 5) when the tumor volume was about 100 mm3. The anti-human TIM3 antibody significantly inhibited tumor growth in homozygous B-hTIM3 mice, suggesting that the B-hTIM3 mouse model is an effective tool for in vivo hTIM3 antibody efficacy studies. (A) Tumor average volume ± SEM, (B) Mice average weight ± SEM.


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