B-hVEGFR2 mice
| Strain Name |
C57BL/6-Kdrtm1(KDR)Bcgen/Bcgen
|
Common Name | B-hVEGFR2 mice |
| Background | C57BL/6 | Catalog number | 110113 |
|
Related Genes |
CD309, FLK1, VEGFR, KDR | ||
|
NCBI Gene ID |
16542 | ||
mRNA expression analysis
Strain specific analysis of VEGFR2 gene expression in wild type and B-hVEGFR2 mice by RT-PCR. Mouse Vegfr2 mRNA was detectable only in embryo of wild type (+/+) mice. Human VEGFR2 mRNA was detectable only in homozygous B-hVEGFR2 (H/H) mice but not in wild type mice.
Protein expression analysis
Strain specific VEGFR2 expression analysis in homozygous B-hVEGFR2 mice by flow cytometry. Embryonic lung endothelial cells were collected from wild type (+/+) and homozygous B-hVEGFR2 (H/H) mice and analyzed by flow cytometry with anti-VEGFR2 antibody. Anti-Mouse VEGFR2 can cross-react with human/mouse VEGFR2 protein. Combined with mRNA expression detection in wild-type and homozygous B-hVEGFR2 mice This could prove that mouse VEGFR2 was detectable in wild type mice . Human VEGFR2 was exclusively detectable in homozygous B-hVEGFR2 but not in wild type mice.
Analysis of spleen leukocytes cell subpopulations in B-hVEGFR2 mice
Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hVEGFR2 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 the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized 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 by FACS. Splenocytes were isolated from female C57BL/6 and B-hVEGFR2 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 TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the spleen. Values are expressed as mean ± SEM.
Analysis of lymph nodes leukocyte subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hVEGFR2 mice (n=3, 6-week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized does not change the overall development, differentiation or distribution of these cell types in the lymph node. Values are expressed as mean ± SEM.
Analysis of lymph nodes T cell subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hVEGFR2 mice (n=3, 6-week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the lymph node. Values are expressed as mean ± SEM.
Analysis of blood leukocyte subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hVEGFR2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized does not change the overall development, differentiation or distribution of these cell types in the blood. Values are expressed as mean ± SEM.
Analysis of blood T cell subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hVEGFR2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hVEGFR2 mice were similar to those in the C57BL/6 mice, demonstrating that VEGFR2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the blood. Values are expressed as mean ± SEM.
Antitumor activity of anti-human VEGFR2 antibody in B-hVEGFR2 mice. (A) Ramucirumab (in house) inhibited MC38 tumor growth in B-hVEGFR2 mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hVEGFR2 mice (female, 8-week-old, n=6). Mice were grouped when tumor volume reached approximately 100 mm3, at which time they were treated with Ramucirumab (in house) with doses and schedules in panel A. (B) Body weight changes during treatment. (C) Proportion of CD31-positive cell area of total area analyzed in tumor tissue. As shown in panel A, anti-human VEGFR2 antibody was efficacious in controlling tumor growth in B-hVEGFR2 mice, demonstrating that the B-hVEGFR2 mice provide a powerful preclinical model for in vivo evaluation of anti-human VEGFR2 antibodies. As shown in panel C, anti-human VEGFR2 antibody Ramucirumab inhibits vascular endothelial cell proliferation. Values are expressed as mean ± SEM.
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