The double-humanized CTLA-4 and LAG3 mouse model (hCTLA-4/hLAG3) was generated by intercrossing hCTLA-4 and hLAG3 mice.
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).
The hLAG3 humanized model, developed by Knockin at the mouse Lag3 locus, expresses the full length chimeric LAG3 isoform (human signal peptide, human extracellular domains, and mouse transmembrane and intracellular domains).
Both hCTLA-4 and hLAG3 expressions are regulated by endogenous mouse promoters.
Splenocytes from hCTLA-4/hLAG3 double-homozygous mice were activated with coated αCD3/αCD28 and mIL-2 for 2 days (A, B) or indicated time (C,D). hCTLA-4 and mCTLA-4 expressions were evaluated on A) Tregs (viable, CD3+CD4+CD25+Foxp3+), B) conventional CD4+ (viable, CD3+CD4+Foxp3-), and C, D) CD8+ (viable, CD3+CD8+) T cells.
Splenocytes from wild-type or hCTLA-4/hLAG3 double-homozygous mice were activated with coated αCD3/αCD28 and mIL-2 for 2 days (A, B) or indicated time (C, D). hLAG3 and mLAG3 expression on A) Tregs (viable, CD3+CD4+CD25+Foxp3+), B) conventional CD4+ (viable, CD3+CD4+Foxp3-), and C, D) CD8+ (viable, CD3+CD8+) T cells.