Basic Information
-
Targeting strategy
-
Gene targeting strategy for B-hIL2/hIL2R mice.
The mouse Il2 gene that encodes the full coding sequence was replaced by human IL2 full coding sequence.
The mouse Il2ra gene that encodes the extracellular domain were replaced by human IL2RA counterpart gene sequences.
The mouse Il2rb gene that encodes the extracellular domain were replaced by human IL2RB counterpart gene sequences.
The mouse Il2rg gene that encodes the full coding region sequences were replaced by human IL2RG counterpart gene sequences.
B-hIL2/hIL2R mice was obtained by B-hIL2/hIL2RA/hIL2RB mice cross breeding with B-hIL2RB/hIL2RG mice.
-
Validation data of B-hIL2/hIL2RA/hIL2RB mice and B-hIL2RB/hIL2RG mice
-
Protein expression analysis
Strain specific IL2 expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by ELISA. Serum was collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice stimulated with anti-mCD3 and anti-mCD28 in vivo, and analyzed by ELISA with species-specific IL2 ELISA kit. Mouse IL2 was detectable in wild-type mice. Human IL2 was exclusively detectable in homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice.(ND: Not detectable)
Strain specific IL2RA expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice, and analyzed by flow cytometry with the species-specific anti-IL2RA antibody. Human IL2RA was exclusively detectable in T cells, CD4+ T cells and Treg cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice. However, IL2RA was not detectable in CD8+ T cells in both these two mice.
Strain specific IL2RA expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with the species-specific anti-IL2RA antibody. Human IL2RA was exclusively detectable in T cells, CD4+ T cells, CD8+ T cells and Treg cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice.
Strain specific IL2RB expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice, and analyzed by flow cytometry with the species-specific anti-IL2RB antibody. Human IL2RB was exclusively detectable in T cells, CD4+ T cells and CD8+ T cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice.
-
Validation data of B-hIL2/hIL2R mice
-
Analysis of spleen leukocyte subpopulations by flow cytometry. Splenocytes were isolated from male C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-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 and monocytes decreased in humanized mice, while percent of B cells increased in humanized mice. NK cells, dendritic cells, neutrophils and macrophages in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of spleen T cell subpopulations by flow cytometry. Splenocytes were isolated from female C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-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. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
-
Summary
-
- Protein expression analysis:
Human IL2, IL2RA and IL2RB were detectable in B-hIL2/hIL2RA/hIL2RB mice.
Human IL2RB and IL2RG were detectable in B-hIL2RB/hIL2RG mice.
- Phosphorylation of STAT5 induced by IL2 and IL15/IL15RA complex:
Human IL2 and IL15/IL15RA complex induce pSTAT5 in B-hIL2RB/hIL2RG mice. The IL2RB and IL2RG humanization did not change the signaling pathway.
- Leukocytes cell subpopulation analysis:
- Percent of T cells and monocytes were decreased, while B cells was increased in humanized mice compared to that in C57BL/6 mice.
- NK cells in spleen, blood and lymph node of humanized mice were similar to that in C57BL/6 mice.
- T cell subtypes in thymus, spleen, blood and lymph node of humanized mice were similar to that in C57BL/6 mice.
