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B-hPD-1/hB7-H3 mice
Strain Name C57BL/6-Pdcd1tm1(PDCD1)Cd276tm1(CD276)/Bcgen Common Name  B-hPD-1/hB7-H3 mice
Background C57BL/6 Catalog number  121152
Related Genes 
PD-1, PDCD1, 4Ig-B7-H3, B7-H3, B7H3, B7RP-2

Gene 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. CD276 (Cluster of Differentiation 276) is a human protein encoded by the CD276 gene and belongs to the immunoglobulin superfamily. The transcript of B7-H3 is ubiquitously expressed in normal tissues and solid tumors, the protein is preferentially expressed only in tumor tissues. B7H3 is expressed on immune cells (such as antigen-presenting cells or macrophages) and has inhibitory roles on T cells, contributing to tumor cell immune evasion. Importantly, B7-H3 is highly overexpressed on a wide range of human solid cancers and often correlates with both negative prognosis and poor clinical outcome in patients. Recent studies have shown that B7H3 is a crucial player in tumor growth, invasion, migration, angiogenesis and metastasis beyond the immune regulatory roles. Due to its role in immune evasion, B7-H3 has become an interesting target for new immunotherapeutic treatments. B7-H3 and PD-L1 induce an inhibitory effect on T-cells and change their microenvironment to escape the anti-tumor immune response. B7-H3 and CTLA4 may act synergistically with one another, just as B7-H3 does in the PD-1 pathway. 


Protein expression analysis in T cells

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Strain specific PD-1 expression analysis in homozygous B-hPD-1/hB7-H3  (H/H) mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hB7-H3  (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-PD-1 antibody. Mouse PD-1 was detectable in WT mice. Human PD-1 was exclusively detectable in homozygous B-hPD-1/hB7-H3 but not WT mice.


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Strain specific analysis of B7-H3 expression in WT and B-hPD-1/hB7-H3 mice by western blot. Epididymis were collected from WT (+/+) mice and homozygous B-hPD-1/hB7-H3 mice. Mouse B7-H3 was detectable in WT mice. Human B7-H3 was detectable in the epididymis of B-hPD-1/hB7-H3 mice due to the cross-reactivity of antibodies.


Analysis of spleen leukocyte subpopulations in B-hPD-1/hB7-H3 mice

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Analysis of splenic leukocyte subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hPD-1/hB7-H3 mice (n=3, 6 weeks-old) and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative FACS plots gated on single live CD45+ cells for further analysis. (B) Results of FACS analysis. Percentages of T, B, NK cells, monocytes/macrophages, and DC were similar in homozygous B-hPD-1/hB7-H3 mice and C57BL/6 mice, demonstrating that introduction of hPD-1/hB7-H3 in place of its mouse counterpart does not change the overall development, differentiation, or distribution of these cell types in spleen. Values are expressed as mean ± SEM.


Analysis of spleen leukocyte subpopulations in B-hPD-1/hB7-H3 mice

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Analysis of splenic T cell subpopulations by FACS
Splenocytes were isolated from female C57BL/6 and B-hPD-1/hB7-H3 mice (n=3, 6 weeks-old) and analyzed by flow cytometry for T cell subsets. (A) Representative FACS plots gated on TCRβ+ T cells and further analyzed. (B) Results of FACS analysis. Percentages of CD8+, CD4+, and Treg cells were similar in homozygous B-hPD-1/hB7-H3 and C57BL/6 mice, demonstrating that introduction of hPD-1/hB7-H3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.

Analysis of blood leukocyte subpopulations in B-hPD-1/hB7-H3 mice

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Analysis of blood leukocyte subpopulations by FACS
Blood were isolated from female C57BL/6 and B-hPD-1/hB7-H3 mice (n=3, 6 weeks-old) and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative FACS plots gated on single live CD45+ cells for further analysis. (B) Results of FACS analysis. Percentages of T, B, NK cells, monocytes/macrophages, and DC were similar in homozygous B-hPD-1/hB7-H3 mice and C57BL/6 mice, demonstrating that introduction of hPD-1/hB7-H3 in place of its mouse counterpart does not change the overall development, differentiation, or distribution of these cell types in blood. Values are expressed as mean ± SEM.

Analysis of blood leukocyte subpopulations in B-hPD-1/hB7-H3 mice

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Analysis of blood T cell subpopulations by FACS
Blood were isolated from female C57BL/6 and B-hPD-1/hB7-H3 mice (n=3, 6 weeks-old) and analyzed by flow cytometry for T cell subsets. (A) Representative FACS plots gated on TCRβ+ T cells and further analyzed. (B) Results of FACS analysis. Percentages of CD8+, CD4+, and Treg cells were similar in homozygous B-hPD-1/hB7-H3 and C57BL/6 mice, demonstrating that introduction of hPD-1/hB7-H3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.


Combination therapy of anti-human PD-1 antibody and anti-human B7-H3 antibody


Antitumor activity of Tislelizumab combined with Enoblituzumab in B-hPD-1/hB7-H3 mice. (A) Anti-human PD-1 antibody Tislelizumab combined with anti-human B7-H3 antibody Enoblituzumab (both of the antibodies were produced in house) inhibited B-CAG-hB7-H3 EL4 cells tumor growth in B-hPD-1/hB7-H3 mice. B-CAG-hB7-H3 EL4 cells were subcutaneously implanted into homozygous B-hPD-1/hB7-H3 mice (female, 6-8 week-old, n=6). Mice were grouped when tumor volume reached approximately 90-100 mm3, at which time they were treated with hPD-1 and hB7-H3 antibodies. (B) Body weight changes during treatment. As shown in panel A, combination of hPD-1 and hB7-H3 antibodies were more efficacious in controlling tumor growth in B-hPD-1/hB7-H3 mice. B-hPD-1/hB7-H3 mice provide a powerful preclinical model for in vivo evaluation of anti-human PD-1 and B7-H3 antibodies. Values are expressed as mean ± SEM.