Basic Information
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Targeting strategy
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B-hIL2RB/hIL2RG/hIL15/hIL15RA mice was obtained by crossing B-hIL2RB/hIL2RG mice with B-hIL15/hIL15RA mice.
The targeting strategy of each gene are as follows.
- Exons 2~8 of the mouse Il2rb gene, which encodes the extracellular domain, were replaced by human IL2RB counterpart gene sequences.
- Exons 1~8 of the mouse Il2rg gene that encodes the full coding region sequences were replaced by human IL2RG exons 1~8.
- The mouse Il15 gene that encodes the full coding sequence was replaced by human IL15 counterpart gene sequences.
- The mouse Il15ra gene that encodes the extracellular region was replaced by human IL15RA counterpart gene sequences.
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Protein expression analysis of T cells
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Strain specific IL2RB and IL2RG expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG/hIL15/hIL15RA mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB and IL2RG were detectable in T cells of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
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Protein expression analysis of CD4+ T cells
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Strain specific IL2RB and IL2RG expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG/hIL15/hIL15RA mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB and IL2RG were detectable in CD8+ T cells of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
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Protein expression analysis of CD8+ T cells
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Strain specific IL2RB and IL2RG expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG/hIL15/hIL15RA mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB and IL2RG were detectable in NK cells of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
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Protein expression analysis of NK cells
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Strain specific IL2RB and IL2RG expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG/hIL15/hIL15RA mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB and IL2RG were detectable in NK cells of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
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Protein expression analysis
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Strain specific IL15 expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by ELISA. (A) Mouse IL15 and IL15RA complex; (B) Human IL15; Serum was collected from wild-type mice (+/+) and homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice (H/H), and analyzed by ELISA. Human IL15 was only detectable in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice but not in wild-type mice. Mouse IL15/IL15RA complex was only detectable in wild-type mice but not in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
IL15RA expression analysis in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice by flow cytometry. Dendritic cells were collected from wild-type mice (+/+) and homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice (H/H), and analyzed by flow cytometry with species-specific anti-IL15RA antibody. Mouse IL15RA was detectable in wild-type mice but not in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice. Human IL15RA was exclusively detectable in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice but not in wild-type mice.
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Summary
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Protein expression analysis:
Human IL2RB and IL2RG were detectable in T cells and NK cells of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
Human IL15 was only detectable in serum of homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
Human IL15RA was exclusively detectable in homozygous B-hIL2RB/hIL2RG/hIL15/hIL15RA mice.
Leukocytes cell subpopulation analysis：
The humanization of these genes does not change the overall development, differentiation or distribution of immune cell types in spleen, blood and lymph node.
