B-hTIM3 mice

Basic Information

Targeting Strategy

Gene targeting strategy for B-hTIM3 mice. The exon 2 of mouse Tim3 gene that encode the Ig-like V-type domain were replaced by human TIM3 exon 2 in B-hTIM3 mice.

Details

Phenotype

mRNA expression analysis

Strain specific analysis of TIM3 gene expression in WT and hTIM3 mice by RT-PCR. Mouse Tim3 mRNA was detectable in splenocytes of wild-type (+/+) mice. Human TIM3 mRNA was detectable only in H/H but not in +/+ mice.

Protein Expression Analysis

Strain specific TIM3 expression analysis in homozygous B-hTIM3 mice by flow cytometry. Cells in ascites were collected from WT and homozygous B-hTIM3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-TIM3 antibody. Mouse Tim3 was exclusively detectable in non-T and non-B cells from WT mice. Human TIM3 was exclusively detectable in non-T and non-B cells from homozygous B-hTIM3 but not WT mice.

Strain specific TIM3 expression analysis in homozygous B-hTIM3 mice by flow cytometry.

Splenocytes were collected from WT and homozygous B-hTIM3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-TIM3 antibody. Mouse Tim3 was exclusively detectable in WT mice. Human TIM3 were exclusively detectable in homozygous B-hTIM3 but not WT mice.

Application

In vivo efficacy of anti-human TIM3 antibodies

Antitumor activity of anti-human TIM3 antibodies in B-hTIM3 mice.

 (A) Anti-human TIM3 antibodies inhibited MC38 tumor growth in B-hTIM3 mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hTIM3 mice (male, 7 week-old, n=5). Mice were grouped when tumor volume reached approximately 100 mm³, at which time they were treated with anti-human TIM3 antibody with doses and schedules indicated in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human TIM3 antibody was efficacious in controlling tumor growth in B-hTIM3 mice, demonstrating that the B-hTIM3 mice provide a powerful preclinical model for in vivo evaluation of anti-human TIM3 antibodies. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human TIM3 antibodies

Antitumor activity of anti-human TIM3 antibodies in B-hTIM3 mice.

(A) Anti-human TIM3 antibodies inhibited MC38 tumor growth in B-hTIM3 mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hTIM3 mice (female, 4 week-old, n=5). Mice were grouped when tumor volume reached approximately 100 mm³, at which time they were treated with two anti-human TIM3 antibodies with doses and schedules indicated in panel (B) Body weight changes during treatment. As shown in panel A, anti-human TIM3 Ab2 was efficacious in controlling tumor growth in B-hTIM3 mice, demonstrating that the B-hTIM3 mice provide a powerful preclinical model for in vivo evaluation of anti-human TIM3 antibodies. Values are expressed as mean ± SEM.

Publications using B-hTIM3 mice

1. Zhang D, Jiang F, Zaynagetdinov R, Huang H, Sood VD, Wang H, Zhao X, Jenkins MH, Ji Q, Wang Y, Nannemann DP, Musil D, Wesolowski J, Paoletti A, Bartholomew T, Derner MG, An Q, Iffland C, Halle JP. Identification and characterization of M6903, an antagonistic anti-TIM-3 monoclonal antibody. Oncoimmunology. 2020 Apr 1;9(1):1744921. doi: 10.1080/2162402X.2020.1744921. PMID: 32313722; PMCID: PMC7153820.

References

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4. J Immunol. 2010 Feb 15;184(4):1918-30. doi: 10.4049/jimmunol.0903059. Epub 2010
Jan 18.