B-hPD-1/hPD-L1 mice

Basic Information

Strain Name
C57BL/6-Pdcd1tm1(PDCD1) Cd274tm1(CD274)/Bcgen
Stock Number
120522
Common Name
B-hPD-1/hPD-L1 mice
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
PDCD1 (Programmed death-1,as known as PD-1) ; CD274 (Programmed cell death ligand-1,also known as B7-H1, PD-L1)
Species
C57BL/6
Appearance
Black
Genotypes
Homozygous

Description

PD-1 (Programmed death-1) is mainly expressed on the surface of T cells and primary B cells. The two PD-1 ligands, PD-L1 and PD-L2, are widely expressed on antigen-presenting cells (APCs). PD-1 interacts with its ligands and plays an important role in the negative regulation of the immune response. PD-L1 protein expression is detected in many human tumor tissues. PD-L1 expression in tumor cells could be induced by the microenvironment of tumor cells. PD-L1 expression is favorable for tumorigenesis and growth, for induction of anti-tumor T Cell Apoptosis, and for escaping responses by the immune system. Inhibition of PD-1 binding to its ligand can result in tumor cells that are exposed to the killing version of the immune system, and thus is a target for cancer treatments.

PD-L1 (Programmed cell death ligand-1), also known as B7-H1 and CD274, is mainly expressed in antigen-presenting cells (APCs) and activated T cells, and is one of the two ligands of PD-1. The interaction between PD1 and PD-L1 plays an important role in the negative regulation of the immune response. PD-L1 is highly expressed in a variety of solid tumors. PD-1 and PD-L1 interactions can reduce T Cell Activation and promote tumor immune escape. The PD-1/PD-L1 signaling pathway can be blocked and antitumor immune response can be restored by using by anti-PD-1 or anti-PD-L1 antibodies to block the binding of PD1 to PD-L1.

Targeting Strategy

Gene targeting strategy for B-hPD-1/hPD-L1 mice. The exon 2 of mouse PD-1 gene that encode the extracellular domain was replaced by human PD-1 exon 2 in B-hPD-1/hPD-L1 mice. The exon 3 of mouse Pdl1 gene that encode the extracellular domain was replaced by human PD-L1 exon 3 in B-hPD-1/hPD-L1 mice. This double knock-in model, was developed by breeding the B-hPD-1 mice and the B-hPD-L1 mice.

Details

Phenotype

mRNA Expression Analysis

B-hPD-1/PD-L1 Mice

Strain specific analysis of PD-1 and PD-L1 gene expression in WT and B-hPD-1/hPD-L1 mice by RT-PCR. Mouse Pd-1 and PD-L1 mRNA was detected in splenocytes of wild-type (+/+) mice. Human PD-1 and PD-L1 mRNA was detected in H/H but not in +/+ mice.

Protein Expression Analysis

Strain specific PD1 and PD-L1 expression analysis in homozygous B-hPD-1/hPD-L1 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hPD-L1 (H/H) mice stimulated with anti-CD3ε in vivo , and analyzed by flow cytometry with species-specific anti-PD-1 and anti-PD-L1 antibodies. Mouse PD-1 and PD-L1 were exclusively detectable in WT mice. Human PD-1 and PD-L1 were exclusively detectable in homozygous B-hPD-1/hPD-L1 mice but not in WT mice.

Blood routine test in B-hPD-1/hPD-L1 mice

Complete blood count (CBC)

Blood from female C57BL/6 and B-hPD-1/hPD-L1 mice (n=5, 6 weeks-old) was collected and analyzed by CBC. There was no differences among any measurement between C57BL/6 and B-hPD-1/hPD-L1 mice.

Blood chemistry of B-hPD-1/hPD-L1 mice

Blood chemistry tests of B-hPD-1/hPD-L1 mice.Serum from the C57BL/6 and B-hPD-1/hPD-L1 mice (n=3, 6 week-old) was collected and analyzed for levels of biochemistry. There was no differences on AST, ALB, GLU, CHOL, CREA measurement between C57BL/6 and B-hPD-1/hPD-L1. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human PD-L1 antibody

Antitumor activity of anti-human PD-L1 antibody in B-hPD-1/hPD-L1 mice. (A) Anti-human PD-L1 antibody atezolizumab inhibited MC38-hPD-L1 tumor growth in B-hPD-1/hPD-L1 mice. Murine colon cancer MC38-hPD-L1 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1/hPD-L1 mice (male, 5-week-old, n=5). Mice were grouped when tumor volume reached approximately 100 mm3, at which time they were treated with anti-human PD-L1 antibody with doses and schedules indicated in panel. (B) Body weight changes during treatment. As shown in panel A, different doses of human PD-L1 antibody inhibited tumor growth in different levels, demonstrating that the B-hPD-1/hPD-L1 mice provide a powerful preclinical model for in vivo evaluation of anti-human PD-L1 antibodies. Values are expressed as mean ± SEM.

Combination therapy of anti-human PD-1 Ab and anti-human PD-L1 Ab

Antitumor activity of anti-human PD-1 antibody pembrolizumab combined with anti-human PD-L1 antibody atezolizumab in B-hPD-1/hPD-L1 mice. (A) Pembrolizumab combined with atezolizumab inhibited MC38-hPD-L1 tumor growth in B-hPD-1/hPD-L1 mice. Murine colon cancer MC38-hPD-L1 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1/hPD-L1 mice mice (female, 5-7 weeks old, n=6). Mice were grouped when tumor volume reached approximately 150 mm3, at which time they were treated with Pembrolizumab and atezolizumab with doses and schedules indicated in panel. (B) Body weight changes during treatment. As shown in panel A, pembrolizumab alone gives anti-tumor effect, low dose of pembrolizumab combination of atezolizumab did not show significant difference compared with atezolizumab alone at tested condition. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human PD-1 antibody

Antitumor activity of anti-human PD-1 antibody in B-hPD-1/hPD-L1 mice. (A) Anti-human PD-L1 antibody Pembrolizumab (in house) inhibited MC38-hPD-L1 tumor growth in B-hPD-1/hPD-L1 mice. Murine colon cancer MC38-hPD-L1 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1/hPD-L1 mice (male, 8-week-old, n=5). Mice were grouped when tumor volume reached approximately 80mm3, at which time they were treated with anti-human PD-1 antibody with doses and schedules indicated in panel. (B) Body weight changes during treatment. As shown in panel A, different doses of human PD-1 antibody inhibited tumor growth in different levels, demonstrating that the B-hPD-1/hPD-L1 mice provide a powerful preclinical model for in vivo evaluation of anti-human PD-1 antibodies. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human PD-L1 antibody

Antitumor activity of anti-human PD-L1 antibody in B-hPD-1/hPD-L1 mice. (A) Anti-human PD-L1 antibody atezolizumab (in house) inhibited MC38-hPD-L1 tumor growth in B-hPD-1/hPD-L1 mice. Murine colon cancer MC38-hPD-L1 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1/hPD-L1 mice (male, 8-week-old, n=5). Mice were grouped when tumor volume reached approximately 80mm3, at which time they were treated with anti-human PD-L1 antibody with doses and schedules indicated in panel. (B) Body weight changes during treatment. As shown in panel A, different doses of human PD-L1 antibody inhibited tumor growth in different levels, demonstrating that the B-hPD-1/hPD-L1 mice provide a powerful preclinical model for in vivo evaluation of anti-human PD-L1 antibodies. Values are expressed as mean ± SEM.

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