Basic Information
-
Targeting Strategy
-
Gene targeting strategy for B-hCTLA4/hTIM3 mice. In B-hCTLA4 mice, exon 2 of mouse Ctla4 (which encodes the extracellular domain) was replaced by human CTLA4 exon 2. In B-hTIM3 mice, exon 2 of mouse Tim3 (which encodes the extracellular domain) was replaced by human TIM3 exon 2. The B-hCTLA4/hTIM3 double knock-in model was developed by breeding B-hCTLA4 mice and the B-hTIM3 mice. These B-hCTLA4/hTIM3 mice have a functional mouse immune system.
-
Expression Analysis
-
Protein expression analysis
Strain specific TIM3 expression analysis in homozygous B-hCTLA4/hTIM3 mice by flow cytometry.
Splenocytes were collected from WT and homozygous B-hCTLA4/hTIM3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-TIM3 antibody. Mouse TIM3 was detectable in WT mice. Human TIM3 was exclusively detectable in homozygous B-hCTLA4/hTIM3 but not WT mice.
Strain specific CTLA4 expression analysis in homozygous B-hCTLA4/hTIM3 mice by flow cytometry.
Splenocytes were collected from WT and homozygous B-hCTLA4/hTIM3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-CTLA4 antibody. Mouse CTLA4 was detectable in WT mice. Human CTLA4 was exclusively detectable in homozygous B-hCTLA4/hTIM3 but not WT mice.
-
References
-
- Cell Research (2018) 0:1–15; doi: 10.1038/s41422-018-0012-z
- Cancer Immunity (22 January 2013) Vol. 13, p. 5
- Journal of Immunological Methods. 18 November 2016. Doi: 10.1016/j.jim.2017.04.006
