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B-hPD-1/hLAG3 mice
Strain Name C57BL/6-Pdcd1tm1(PDCD1)Lag3tm1(LAG3)/Bcgen Common Name  B-hPD-1/hLAG3 mice
Background C57BL/6 Catalog number  120520
Related Genes 
PD-1 (Programmed death-1) ;
LAG3
(Lymphocyte activation gene 3, CD223)

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-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 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. LAG3 (Lymphocyte activation gene 3, CD223) is a lymphocyte activation gene and a member of the Ig family. It is mainly expressed in activated T cells, NK cells, B cells and plasmacytoid DCs. LAG3 is a negative regulator of immunity and mainly binds to MHC class Ⅱ molecules to regulate the function of dendritic cells. The expression of LAG3 is associated with the negative immunoregulatory function of specific T cells. Inhibition of LAG3 function enhances the antitumor effect of specific CD8+ T cells, therefore LAG3 is a potential target for tumor immunotherapy.


Protein expression analysis




Strain specific PD-1 and LAG3 expression analysis in homozygous B-hPD-1/hLAG3 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hLAG3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-PD-1 and anti-LAG3 antibody. Mouse PD-1 and LAG3 were detectable in WT mice while mouse LAG3 was also detectable in homozygous B-hPD-1/hLAG3, due to the anti-mouse LAG3 antibody cross-reacts with human LAG3. Human PD-1 and LAG3 were exclusively detectable in homozygous B-hPD-1/hLAG3 but not WT mice.



Analysis of spleen leukocytes cell subpopulations in B-hPD-1/hLAG3 mice

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Analysis of spleen leukocyte subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 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, B, NK, Monocyte, DC and macrophage cells in homozygous B-hPD-1/hLAG3 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen.



Analysis of spleen T cell subpopulations in B-hPD-1/hLAG3 mice

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Analysis of spleen T cell subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 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/hLAG3 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in spleen. Values are expressed as mean ± SEM.


Analysis of blood leukocytes cell subpopulations in B-hPD-1/hLAG3 mice

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Analysis of blood leukocyte subpopulations by FACS

Blood were isolated from female C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 week-old) Flow cytometry analysis of the blood leukocytes 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, B, NK, Monocyte, DC and macrophage cells in homozygous B-hPD-1/hLAG3 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen.



Analysis of blood T cell subpopulations in B-hPD-1/hLAG3 mice

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Analysis of blood T cell subpopulations by FACS

Blood were isolated from female C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 week-old).  Flow cytometry analysis of the blood leukocytes 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/hLAG3 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in spleen. Values are expressed as mean ± SEM.




Blood routine test in B-hPD-1/hLAG3 mice

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Complete blood count (CBC). Blood from female C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 week-old) was collected and analyzed for CBC. There was no differences among any measurement between C57BL/6 and B-hPD-1/hLAG3 mice, indicating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change blood cell composition and morphology. Values are expressed as mean ± SEM.


Blood chemistry of B-hPD-1/hLAG3 mice

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Blood chemistry tests of B-hPD-1/hLAG3 mice. Serum from the C57BL/6 and B-hPD-1/hLAG3 mice (n=3, 6 week-old) was collected and analyzed for levels of ALT and AST. There was no differences on either measurement between C57BL/6 and B-hPD-1/hLAG3 mice, indicating that introduction of hPD-1/hLAG3 in place of its mouse counterpart does not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.




Combination therapy of anti-human PD-1 antibody and anti-human LAG3 antibody



Antitumor activity of anti-human PD-1 antibody combined with anti-human LAG3 antibody in B-hPD-1/hLAG3 mice. (A) Anti-human PD-1 antibody combined with anti-human LAG3 antibody inhibited MC38 tumor growth in B-hPD-1/hLAG3 mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hPD-1/hLAG3 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 pembrolizumab and anti-human LAG3 antibody with doses and schedules indicated in panel (B) Body weight changes during treatment. As shown in panel A, combination of pembrolizumab and anti-hLAG3 antibody shows more inhibitory effects than individual groups, demonstrating that the B-hPD-1/hLAG3 mice provides a powerful preclinical model for in vivo evaluating combination therapy efficacy of hPD-1 antibodies and hLAG3 antibodies. Values are expressed as mean ± SEM.