B-hPD-1/hPD-L1/hTNFR2 mice

Basic Information

Strain Name
C57BL/6-Pdcd1tm1(PDCD1)Cd274tm1(CD274)Tnfrsf1btm1(TNFRSF1B)/Bcgen
Stock Number
130849
Common Name
B-hPD-1/hPD-L1/hTNFR2 mice
Background
C57BL/6
Related Genes
PD-1 (Programmed death-1) ; CD274 (CD274 antigen) TNFRSF1B (Tumor necrosis factor receptor superfamily, member 1b)

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-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.

TNFR2 is a member of the tumor necrosis factor receptor superfamily, which also contains TNFRSF1A. This protein and TNF-receptor 1 form a heterodimer that mediates the recruitment of two anti-apoptotic proteins, c-IAP1 and c-IAP2, which possess E3 ubiquitin ligase activity. The function of IAPs in TNF-receptor signaling is unknown, however, c-IAP1 is thought to potentiate TNF-induced apoptosis by the ubiquitination and degradation of TNF-receptor-associated factor 2 (TRAF2), which mediates anti-apoptotic signals. In the tumor microenvironment, TNFR2 antagonism is thought to activate effector T cells, resulting in tumor cell death.

Targeting Strategy

The exon 2 of mouse Pd-1 gene that  encodes the extracellular domain was replaced by human PD-1 exon 2 in B-hPD-1/hPD-L1/hTNFR2 mice. The exon 3 of mouse Pd-l1 gene that  encodes the extracellular domain was replaced by human PD-L1 exon 3 in B-hPD-1/hPD-L1/hTNFR2 mice. The exons 2~6 of mouse Tnfr2 gene that  encode the extracellular domain was replaced by human TNFR2 exons 2~6 in B-hPD-1/hPD-L1/hTNFR2 mice.

Details

Protein expression analysis in Tregs

PD-1, PD-L1 and TNFR2 expression analysis in homozygous B-hPD-1/hPD-L1/hTNFR2 (H/H) mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hPD-L1/hTNFR2 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-PD-1, anti-PD-L1 and anti-TNFR2 antibodies. Mouse PD-1, PD-L1 and TNFR2 were detectable in WT mice. Human PD-1, PD-L1 and TNFR2 were exclusively detected in homozygous B-hPD-1/hPD-L1/hTNFR2 but not WT mice.

Analysis of blood, spleen and lymph node leukocyte subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice

Analysis of blood, spleen and lymph node leukocytes cell subpopulations by FACS

Blood, spleen and lymph node leukocytes cell were isolated from female mice in the panel(n=3, 6 week-old). Flow cytometry analysis was performed to assess leukocyte subpopulations. Percent of T, B, NK, Granulocytes, Monocyte, DC and macrophage cells in homozygous B-hPD-1/hPD-L1/hTNFR2 mice were similar to those in C57BL/6 mice, demonstrating that the humanized mouse does not change the overall development, differentiation or distribution of these cell types in blood, spleen and lymph node.

Analysis of blood, spleen and lymph node T cell subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice

Analysis of blood, spleen and lymph node T cell subpopulations by FACS

Blood, spleen and lymph node leukocytes cell were isolated from female mice in the panel(n=3, 6 week-old). Flow cytometry analysis was performed to assess leukocyte subpopulations. Percentages of CD4+T, CD8+T and Foxp3+ Treg cells in homozygous B-hPD-1/hPD-L1/hTNFR2 mice were similar to those in C57BL/6 mice, demonstrating that the humanized mouse does not change the overall development, differentiation or distribution of these cell types in blood, spleen and lymph node.

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