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Design of the hPD‑1/hCTLA-4 mouse
The double-humanized PD-1 and CTLA-4 mouse model (hPD-1/hCTLA-4) was generated by intercrossing hPD-1 and hCTLA-4 mice.
hPD-1 has been developed by inserting, within the mouse PD-1 locus, a chimeric PD-1 with a human extracellular domain, a murine transmembrane domain and a murine intracellular domain. The design of hCTLA-4, developed by Knockin at the mouse CTLA-4 locus, enables the expression of: a) the full-length chimeric CTLA-4 isoform (human extracellular domain and mouse intracellular domain); and b) the soluble isoform (human extracellular domain).
Both hPD-1 and hCTLA-4 expressions are regulated by endogenous mouse promoters.
hPD-1/hCTLA-4 features- The PD-1 and CTLA-4 extracellular domains are entirely humanized
- Physiological regulation and expression pattern of the human PD-1 and CTLA-4
- Preservation of the target-ligand interaction
- Fully functional mouse immune system
- Lack of expression of the murine target gene, thus avoiding cross-reactivity
Splenocytes from hPD-1/hCTLA-4 double homozygous mice were activated with αCD3/αCD29. Expressions of hPD-1 and mPD-1 (upper panel), and hCTLA and mCTLA-4 (lower panel) were evaluated on conventional CD4 (viable, CD3+CD4+Foxp3-) T cells at indicated time points.
PD-1 and CTLA-4 expression on TILs from mice bearing MCA205 tumors. Mouse and human PD-1 and CTLA-4 expression were analyzed on wild-type and hPD-1/hCTLA-4 homozygous mice.
