B-hPD-1 mice Plus

Basic Information

Strain Name
C57BL/6-Pdcd1tm3(PDCD1)Bcgen/Bcgen
Stock Number
110019
Common Name
B-hPD-1 mice Mice plus
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
Pd-1 (Programmed death-1)
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 tumor escape from 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.

Targeting Strategy

Details

Phenotype

Protein Expression Analysis

Strain specific PD-1 expression analysis in homozygous B-hPD-1 mice plus by flow cytometry.

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

Strain specific PD-1 expression analysis in homozygous B-hPD-1 mice plus by flow cytometry.

Splenocytes were collected from WT and homozygous B-hPD-1 mice plus (H/H) and analyzed by flow cytometry with species-specific anti-PD-1 antibody. Mouse PD-1 was detected in WT mice. Human PD-1 was exclusively detected in homozygous B-hPD-1 mice plus but not WT mice.

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

Analysis of spleen leukocyte subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hPD-1 mice plus (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, monocytes, DCs, granulocytes and macrophages in homozygous B-hPD-1 mice plus mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1 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 T cell subpopulations in B-hPD-1 mice plus

Analysis of spleen T cell subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hPD-1 mice plus (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8, CD4, and Treg cells in homozygous B-hPD-1 mice plus were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1 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 lymph node T cell subpopulations in B-hPD-1 mice plus

Analysis of lymph node T cell subpopulations by FACS

lymph node T were isolated from female C57BL/6 and B-hPD-1 mice plus (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of NK, CD8, CD4, and Treg cells in homozygous B-hPD-1 mice plus were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1 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.

T cells in B-hPD-1 plus mice bind anti-human PD-1 antibody

Analysis of splenocytes of B-hPD-1 mice plus by FACS.

Splenocytes were isolated from female B-hPD-1 mice plus. Flow cytometry analysis of the splenocytes was performed to assess human PD-1 expression on T cells. Single live cells were gated for CD45 population and used for further analysis as indicated here. Human PD-1 expression was detectable on T cells in B-hPD-1 mice plus as evidenced by Nivolumab and Pembrolizumab binding vs isotype control.

Application

In vivo efficacy of anti-human PD-1 antibody

Antitumor activity of anti-human PD-1 antibody nivolumab in B-hPD-1 mice plus.

(A) Anti-human PD-1 antibody inhibited MC38 tumor growth in B-hPD-1 mice plus. Murine colon cancer MC38 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1 mice plus (male 8-week-old, n=5). Mice were grouped when tumor volume reached approximately 150 mm3, 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, anti-human PD-1 antibody was efficacious in controlling tumor growth in B-hPD-1 mice plus, demonstrating that the B-hPD-1 mice plus 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-1 antibodies

Antitumor activity of anti-human PD-1 antibodies Pembrolizumab in B-hPD-1 mice plus.

(A) Anti human PD-1 antibodies inhibited MC38 tumor growth in B-hPD-1 mice plus. Murine colon cancer MC38 cells (5×105) were subcutaneously implanted into homozygous B-hPD-1 mice plus (male 8-week-old, n=5). Mice were grouped when tumor volume reached approximately 150 mm3, at which time they were treated with three anti-human PD-1 antibodies with doses and schedules indicated in panel (B) Body weight changes during treatment. As shown in panel A, anti-human PD-1 antibodies were efficacious in controlling tumor growth in B-hPD-1 mice plus, demonstrating that the B-hPD-1 mice plus provide a powerful preclinical model for in vivo evaluation of anti-human PD-1 antibodies. Values are expressed as mean ± SEM.

References

1. Nat Commun. 2017 Feb 6;8:14369. doi: 10.1038/ncomms14369.
2. EMBO J. 1992 Nov;11(11):3887-95.
3. J Exp Med. 2000 Oct 2;192(7):1027-34.
4. Science. 2001 Jan 12;291(5502):319-22.

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