Basic Information

Strain Name
C57BL/6-Nt5etm1(NT5E)/Bcgen
Stock Number
110027
Common Name
B-hCD73 mice
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
NT5E (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 homozygous B-hCD73 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice.

 

Strain specific CD73 expression analysis in homozygous B-hCD73 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice.

 

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

 

Strain specific CD73 expression analysis in homozygous B-hCD73 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hCD73 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-CD73 antibody. Mouse CD73 was detectable in WT mice. Human CD73 was exclusively detectable in homozygous B-hCD73 but not WT mice.

 

Antibody binding assay

Analysis of B-CAG-hCD73 MC38 by FACS. Flow cytometry analysis of the B-CAG-hCD73 MC38 was performed to assess anti-human CD73 Ab. Single live cells were gated and used for further analysis as indicated here. Human CD73 expression was detectable on B-CAG-hCD73 MC38 as evidenced by Hu101-28 (in house) and Oleclumab (in house) binding vs isotype control.

 

Analysis of splenocytes of B-hCD73 mice by FACS. Splenocytes were isolated from female B-hCD73 mice (n=3, 6 week-old). Flow cytometry analysis of the splenocytes was performed to assess anti-human CD73 Ab binding with Treg cells. Single live cells were gated for CD45+Foxp3+ population and used for further analysis as indicated here. Human CD73 expression was detectable on Foxp3 Treg cells in B-hCD73 mice as evidenced by Hu101-28 (in house) and Oleclumab (in house) binding vs isotype control.

 

In vivo efficacy of anti-human CD73 antibodies

Antitumor activity of anti-human CD73 antibodies in B-hCD73 mice. (A) Anti-human CD73 antibodies inhibited hCD73-MC38 tumor growth in B-hCD73 mice. Murine colon cancer hCD73-MC38 cells were subcutaneously implanted into homozygous B-hCD73 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 anti-human CD73 antibodies with doses and schedules indicated in panel (B) Body weight changes during treatment. As shown in panel A, two anti-human CD73 antibodies were efficacious in controlling tumor growth in B-hCD73 mice, demonstrating that the B-hCD73 mice provide a powerful preclinical model for in vivo evaluation of anti-human CD73 antibodies. Values are expressed as mean ± SEM.

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