Basic Information

Strain name
C57BL/6-Pdcd1tm3(PDCD1)Bcgen Cd274tm1(CD274)Bcgen Pdcd1lg2tm3(PDCD1LG2)Bcgen/Bcgen
Common name
B-hPD-1 plus/hPD-L1/hPD-L2 mice
Background
C57BL/6
Catalog number
112710
Aliases
CD279, PD-1, PD1, SLEB2, hPD-1, hPD-l, hSLE1; B7-H, B7H1, PD-L1, PDCD1L1, PDCD1LG1, PDL1, hPD-L1; B7DC, Btdc, CD273, PD-L2, PDCD1L2, PDL2, bA574F11.2
NCBI gene ID
Application
B-hPD-1 plus/hPD-L1/hPD-L2 mice is a powerful mouse model for pharmacodynamics and safety evaluation of drugs targeted PD-1/PD-L1/PD-L2 signaling pathways.

Targeting strategy

Gene targeting strategy for B-hPD-1 plus/hPD-L1/hPD-L2 mice. 

Human PD-1 gene encoding the extracellular region and mouse PD-1 gene encoding the transmembrane and cytoplasmic region were inserted after the initiation codon ATG of mouse PD-1 gene in B-hPD-1 mice plus. The exon 3 of mouse Pdl1 gene that encodes the IgV domain was replaced by human PD-L1 exon 3 in B-hPD-L1 mice. B-hPD-1 plus/hPD-L1 mice was developed by cross-mating the B-hPD-1 mice plus and the B-hPD-L1 mice together. The exons 3-4 of human PD-L2 encoding the extracellular region were knocked-in to replace the exon 3-4 sequences of mouse PD-L2 in B-hPD-1 plus/hPD-L1 mice. Thus, the B-hPD-1 plus/hPD-L1/hPD-L2 mice were obtained.

mRNA expression analysis in humanized B-hPD-1 plus/hPD-L1/hPD-L2 mice

Species specific analysis of PD-L2 gene expression in wild-type C57BL/6 mice and homozygous humanized B-hPD-1 plus/hPD-L1/hPD-L2 mice by RT-PCR. Thymus was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1 plus/hPD-L1/hPD-L2 mice (H/H). Mouse PD-L2 mRNA was detectable only in wild-type C57BL/6 mice. Human PD-L2 mRNA was only detectable in homozygous B-hPD-1 plus/hPD-L1/hPD-L2 mice, but not in wild-type C57BL/6 mice.

 Protein expression analysis in spleen

Strain specific PD-L2 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hPD-1 plus/hPD-L1/hPD-L2 mice by flow cytometry. Bone marrow was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1 plus/hPD-L1/hPD-L2 mice (H/H) and cultured in vitro to induce the bone marrow-derived dendritic cells (BMDCs). The successfully induced BMDCs were stimulated with or without LPS in vitro. Expression of PD-L2 on BMDCs was analyzed by flow cytometry. Mouse PD-L2 was only detectable in wild-type C57BL/6 mice. Human PD-L2 was exclusively detectable in homozygous B-hPD-1 plus/hPD-L1/hPD-L2 mice, but not in wild-type C57BL/6 mice.

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