Immune Checkpoint Mouse Models

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Applied StemCell’s off-shelf mouse catalog offers >100 humanized immune checkpoint mouse models, with single, double, and triple humanized gene models.

  • Knock-in of humanized counterparts of immune checkpoint gene into the endogenous locus.
  • Proprietary human transgene knock-in strategy to enable screening of many different antibodies and immuno-therapy compounds.
  • In vivo and in vitro validated for physiological expression of human version of the genes (in-house and through collaborators).
  • Fully functional murine immune system  
  • The humanized immune checkpoint mice have been generated in the most widely used C57BL/6J background used in immunology and oncology studies.
  • Ideal for immunology and antibody/ immuno-oncology therapy efficacy studies and drug discovery research.

Validation Data for Humanized PD-L1 Mouse

Strain Name: C57BL/6J-Cd274em1(hPD-L1)Asc        Strain Background: C57BL/6J

Programmed cell death 1 ligand (PD-L1), also known as cluster of differentiation 274(CD274) or B7 homolog 1 (B7H1), is a 40kDa transmembrane protein encoded by the gene CD274 in human. The binding of PD-L1 to the PD-1 receptors expressed on the surface of activated T cells transmits a negative regulatory signal. While under normal circumstances the PD-L1 pathway acts as a type of "off switch" that helps keep the T cells from attacking other cells, the high expression level of PD-L1 in the tumor microenvironment inhibits the function of tumor-infiltrating T cells, thereby allowing tumors cells to escape immune surveillance.

Construction Strategy

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Figure 1. Generation strategy of humanized PD-L1 mice. The full-length protein coding sequence for human PD-L1 was placed immediately downstream of the start codon of the mouse endogenous Pd-l1 gene, followed by a poly(A) site, so that the expression of endogenous Pd-l1 in the mouse was replaced by the expression of fully humanized PD-L1 protein. This mouse model was generated in the C57BL/6J genetic background.

Validation Data

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Figure 2. hPD-L1 expression in spleen T cells after stimulation. Flow cytometry analysis of spleen lymphocytes collected from homozygous humanized PD-L1 mice and wild-type C57BL/6J mice. The results showed that the expression of human PD-L1 can be detected in both T cells and B cells collected from the spleen of homozygous humanized PD-L1 mice. (Data provided by collaborators).

 appnote-animalmodel-mouse-humanizedicp-invivo

Figure 3. In vivo validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized PD-L1 mice. Mouse colon cancer cells MC38 engineered to express human PD-L1 were implanted subcutaneously into homozygous, humanized PD-L1 mice. The mice were randomly assigned into two groups when the tumor volume reached 100 mm3, with one group receiving human IgG as a control and the other receiving a human-specific, PD-L1 antibody (n=5). The human PD-L1 blocking antibody significantly inhibited tumor growth in the homozygous humanized mice (Left) without affecting overall body weight (Right), suggesting that the humanized PD-L1 mice represent an ideal model for evaluating the efficacy of therapeutic antibodies targeting human PD-L1. (TGI: tumor growth inhibition; 67%; p < 0.001), demonstrating that the humanized PD-L1 mice are a good in vivo model for validating the efficacy of antibodies targeting human PD-L1.

 


Single Humanized
Immune-checkpoint Mice
 
 Double Humanized
Immune-checkpoint Mice
Triple Humanized
Immune-checkpoint Mice
 
CTLA4  CD81 PD-1 & GITR  PD-1 & PD-L1 & OX40
CTLA4 CEACAM1  PD-L1 & GITR  PD-1 & PD-L1 & LAG3
PD-1 CSF1  LAG3 & CTLA4  PD-1 & PD-L1 & PDCD1LG2
PD-L1 CSF2  OX40 & CTLA4  PD-1 & PD-L1 & IDO1
PD-L1 CSF3  PD-1 & CTLA4  PD-1 & TIGIT & TIM3
GITR CXCR2  PD-1 & LAG3  SIRPA & CD47 & PD-1
OX40 Dpp4  PD-L1 & CTLA4  
OX40L ENPP1  LAG3 & TIM3  
VISTA FcRn  CTLA4 & ICOS    
CD3E GITR  PD-1 & CD27  
BTLA HGF PD-1 & CD47   
CD276 ICOS  PD-1 & NT5E  
CD27 ICOS  PD-L1 & CD27  
CD38 ICOSL  PD-L1 & CD47  
CSF1R IL17F  TIM3 & CEACAM1   
IL6 IL17RA  ICOS & ICOSL   
Nt5e IL1B  PD-1 & TIGIT   
TNFSF15 IL23A  PD-L1 & TIGIT   
VISTA IL2  CD19 & CD3E   
4-1BBL IL2RA  CTLA4 & 4-1BB  
CD4 IL3  IL3 & CSF2   
CD80 IL3  IL6 & IL6R   
CD86 IL4RA  PD-1 & 4-1BB  
IDO1 II5  PD-1 & CD3e   
IL17A IL5RA  PD-1 & CD40  
IL6R IL7  PD-1 & OX40  
OX40 IL9  PD-1 & PD-L1   
PSGL-1 KDR PD-1 & SEMA4D   
TNFRSF25 KLRK1  PD-1 & SIRPA   
VTCN1 LAG3  PD-1 & TIM3   
4-1BB  PCSK9 PD-1 & TLR9   
APOE2 PDCD1LG2  PD-L1 & 4-1BB   
APOE3 PDCD1LG2  PD-L1 & CD40   
APOE4 PVR  PD-L1 & LAG3   
BTLA  SEMA4D  PD-L1 & OX40   
CCR2 SIGLEC15  PD-L1 & SEMA4D   
CCR4 SIRPA  PD-L1 & TIM3   
CCR8 SIRPA  SIRPA & CD47   
CD160 SLAMF7  TLR9 & OX40   
CD19 TIGIT    
CD27 TIGIT     
CD28 TIM3     
CD30 TLR7     
CD36 TLR8    
CD38 TLR9    
CD40 TMEM173    
CD47 TNFRSF1B    
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