B-hCD73 mice plus

Basic Information

Strain Name
C57BL/6-Nt5etm2(NT5E)/Bcgen
Stock Number
110047
Common Name
B-hCD73 mice plus
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
CD73 (Nt5e, 5′ nucleotidase, ecto)
Species
C57BL/6
Appearance
Black
Genotypes
Homozygous

Description

CD73 is expressed on the surface of multiple cells of the immune system including T-cells, B-cells, NK-cells and myeloid-derived suppressor cells. CD73 catalyzes the final hydrolysis of AMP to adenosine following CD39 activity. Increased expression of CD73 has been associated with several tumor signaling pathways stimulated by hypoxyia in the TME. Anti-CD73 therapy in combination with both anti-PD1 and anti-CTLA4 therapy was shown to synergistically reduce syngeneic tumor burden and extend survival via IFNγ-dependent, Th1 CD4+-driven expansion of tumor-specific CD8+ T cells.

Details

Protein expression analysis

Strain specific CD73 expression analysis in heterozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and heterozygous B-hCD73 (H/+) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in heterozygous B-hCD73 and WT mice. Human CD73 was exclusively detectable in heterozygous B-hCD73 but not WT mice.

Strain specific CD73 expression analysis in heterozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and heterozygous B-hCD73 (H/+) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in heterozygous B-hCD73 and WT mice. Human CD73 was exclusively detectable in heterozygous B-hCD73 but not WT mice.

Strain specific CD73 expression analysis in heterozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and heterozygous B-hCD73 (H/+) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in heterozygous B-hCD73 and WT mice. Human CD73 was exclusively detectable in heterozygous B-hCD73 but not WT mice.

mRNA expression analysis

Strain specific analysis of CD73 gene expression in WT and B-hCD73 mice(v2) by RT-PCR. Mouse Cd73 mRNA was detectable in splenocytes of wild-type (+/+) mice. Human CD73 mRNA was detectable only in H/H, but not in +/+ mice.

Protein expression analysis

Strain specific CD73 expression analysis in homozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in and WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice. MEDI9447 (in house) is a cross-reactivity antibody between mouse and human.

Strain specific CD73 expression analysis in homozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in and WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice. MEDI9447 (in house) is a cross-reactivity antibody between mouse and human.

Strain specific CD73 expression analysis in homozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in and WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice. MEDI9447 (in house) is a cross-reactivity antibody between mouse and human.

Strain specific CD73 expression analysis in homozygous B-hCD73 mice plus by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice plus, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in and WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice. MEDI9447 (in house) is a cross-reactivity antibody between mouse and human.

Phenotypic analysis

Analysis of spleen leukocytes cell subpopulations in B-hCD73 mice plus

Analysis of spleen leukocyte subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hCD73 mice(v2) (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-hCD73 mice(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 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-hCD73 mice plus

Analysis of spleen T cell subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hCD73(v2) (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-hCD73(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 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.

Phenotypic analysis

Analysis of lymph node leukocytes cell subpopulations in B-hCD73 mice plus

Analysis of lymph node leukocyte subpopulations by FACS

Leukocytes were isolated from female C57BL/6 and B-hCD73 mice(v2) (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 CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T, B and NK cells in homozygous B-hCD73 mice(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node.

Analysis of lymph node T cell subpopulations in B-hCD73 mice plus

Analysis of lymph node T cell subpopulations by FACS

Leukocytes were isolated from female C57BL/6 and B-hCD73 mice(v2) (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 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-hCD73 mice(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in lymph node. Values are expressed as mean ± SEM.

Phenotypic analysis

Analysis of blood leukocytes cell subpopulations in B-hCD73 mice plus

Analysis of blood leukocyte subpopulations by FACS

Blood were isolated from female C57BL/6 and B-hCD73 mice(v2) (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 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-hCD73 mice(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood.

Analysis of blood T cell subpopulations in B-hCD73 mice plus

Analysis of blood T cell subpopulations by FACS

Blood were isolated from female C57BL/6 and B-hCD73(v2) (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 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-hCD73(v2) were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD73 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in blood. Values are expressed as mean ± SEM.

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