B-hPD-1/hPD-L1/hCD73 plus mice

Basic Information

Strain Name
C57BL/6-Pdcd1tm1(PDCD1)Cd274tm1(CD274)Nt5etm2(NT5E)/Bcgen
Stock Number
131036
Common Name
B-hPD-1/hPD-L1/hCD73 plus mice
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
PD-1 (Programmed death-1) ; CD274 (CD274 antigen) CD73 (Nt5e, 5′ nucleotidase, ecto)
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-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. CD73 is expressed on the surface of multiple cells of the immune system including T-cells, B-cells, NK-cells and myeloid-derived suppressor cells. CD73 catalyzes the final hydrolysis of AMP to adenosine following CD39 activity. Increased expression of CD73 has been associated with several tumor signaling pathways stimulated by hypoxyia in the TME. Anti-CD73 therapy in combination with both anti-PD1 and anti-CTLA4 therapy was shown to synergistically reduce syngeneic tumor burden and extend survival via IFNγ-dependent, Th1 CD4+-driven expansion of tumor-specific CD8+ T cells.

Details

Protein expression analysis

Strain specific PD-1, PD-L1 and CD73 expression analysis in homozygous B-hPD-1/hPD-L1/hCD73 plus mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hPD-L1/hCD73 plus mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-PD-1, anti-PD-L1 and anti-CD73 antibody. Mouse PD-1, PD-L1 and CD73 were detectable in WT mice. Human PD-1, PD-L1 and CD73 were exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD73 plus but not WT mice. MEDI9447 (in house) is a cross-reactivity antibody between mouse and human.

Analysis of blood leukocytes cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus mice

Analysis of spleen leukocytes cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus mice

Analysis of lymph node leukocytes cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus mice

Analysis of blood, spleen and lymph node leukocytes cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus 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/hCD73 plus mice were similar to those in the 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, lymph node T cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus mice

Analysis of blood, spleen and lymph node T cell subpopulations in B-hPD-1/hPD-L1/hCD73 plus 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. Percent of CD4+T, CD8+T and Tre cells in homozygous B-hPD-1/hPD-L1/hCD73 plus mice were similar to those in the 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.

Blood routine test results

Complete blood count (CBC). Blood from C57BL/6 and B-hPD-1/hPD-L1/hCD73 plus mice (n=5, 6 week-old, female) were collected and analyzed for CBC. Any measurement of B-hPD-1/hPD-L1/hCD73 plus mice in the panel were similar to C57BL/6, and there was no differences between male and female mice, indicating that humanized mouse does not change blood cell composition and morphology. Values are expressed as mean ± SEM.

Blood chemistry results

Blood chemistry tests of B-hPD-1/hPD-L1/hCD73 plus mice. Serum from C57BL/6 and B-hPD-1/hPD-L1/hCD73 plus mice (n=5, 6 week-old, female) were collected and analyzed for levels of ALT, AST and other indicators in the panel. There was no differences on either measurement between C57BL/6 and humanized mouse, indicating that humanized mouse does not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.

 

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