B-hPD-1/hPD-L1/hTNFR2 mice_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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동물/세포 모델

동물/세포모델

Humanized Mouse Models

Humanizad Mouse Models

Text

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

Strain Name	C57BL/6-Pdcd1^{tm1(PDCD1)Bcgen}Cd274^{tm1(CD274)Bcgen}Tnfrsf1b^{tm1(TNFRSF1B)Bcgen}/Bcgen	Common Name	B-hPD-1/hPD-L1/hTNFR2 mice
Background	C57BL/6	Catalog number	130849
Related Genes	PD-1 (Programmed death-1) CD274 (CD274 antigen) TNFRSF1B (Tumor necrosis factor receptor superfamily, member 1b)
NCBI Gene ID	18566,60533,21938

Phenotypic analysis

Protein expression analysis in T cells

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Strain specific 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 antibody. Mouse PD-1, PD-L1 and TNFR2 were detectable in WT mice. Human PD-1, PD-L1 and TNFR2 were exclusively detectable in homozygous B-hPD-1/hPD-L1/hTNFR2 but not WT mice.

Protein expression analysis in Treg cells

Analysis of blood leukocytes cell subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice

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Analysis of spleen leukocytes cell subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice

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Analysis of lymph node leukocytes cell subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice

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

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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 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/hTNFR2 mice

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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. Percent of CD4+T, CD8+T and Tre cells in homozygous B-hPD-1/hPD-L1/hTNFR2 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 of B-hPD-1/hPD-L1/hTNFR2 mice

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Complete blood count (CBC). Blood from C57BL/6 and B-hPD-1/hPD-L1/hTNFR2 mice (n=5, 6 week-old, female) were collected and analyzed for CBC. Any measurement of B-hPD-1/hPD-L1/hTNFR2 mice in the panel were similar to C57BL/6, indicating that humanized mouse does not change blood cell composition and morphology. Values are expressed as mean ± SEM.

Blood chemistry of B-hPD-1/hPD-L1/hTNFR2 mice

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Blood chemistry tests of B-hPD-1/hPD-L1/hTNFR2 mice. Serum from C57BL/6 and B-hPD-1/hPD-L1/hTNFR2 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.

Combination therapy of anti-human PD-1 and anti-human TNFR2 antibody

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Antitumor activity of anti-human PD-1 antibody combined with anti-human TNFR2 antibody in B-hPD-1/hPD-L1/hTNFR2 mice. (A) Anti-human PD-1 antibody combined with anti-human TNFR2 antibody inhibited hPD-L1 MC38 tumor growth B-hPD-1/hPD-L1/hTNFR2 mice. All antibodies used in the experiments were prepared in-house. hPD-L1 MC38 cells were subcutaneously implanted into homozygous B-hPD-1/hPD-L1/hTNFR2 mice (female, 6-7-week-old, n=5). Mice were grouped when tumor volume reached approximately 100 mm³, at which time they were treated with human PD-1 antibody and human hTNFR2 antibodies (in house) with doses and schedules indicated in panel A. (B) Body weight changes during treatment. As shown in panel A, combination of PD-1 and hTNFR2 antibodies shows more inhibitory effects than individual groups, demonstrating that B-hPD-1/hPD-L1/hTNFR2 mice provide a powerful preclinical model for in vivo evaluating combination therapy efficacy of hTNFR2 antibodies and hPD-1 antibodies. Values are expressed as mean ± SEM.