Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD2 mice.
The exons 1-4 of mouse Cd2 gene that encode the extracellular domain were replaced by human CD2 exons 1-4 in B-hCD2 mice.
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mRNA expression analysis
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Species-specific CD2 gene expression analysis in wild-type and humanized B-hCD2 mice by RT-PCR. Murine Cd2 mRNA was detected in splenocytes isolated from wild-type (+/+) mice, while human CD2 mRNA was exclusively detected in homozygous B-hCD2 (H/H) mice.
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Protein expression analysis
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Protein expression analysis in T cells
Species-specific CD2 protein expression analysis in humanized B-hCD2 mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hCD2 (H/H) mice, and analyzed by flow cytometry using species-specific anti-CD2 antibodies. Murine CD2 protein was detected in wild-type mice, while human CD2 protein was exclusively detected in B-hCD2 mice.
Protein expression analysis in B cells
Species-specific CD2 protein expression analysis in humanized B-hCD2 mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hCD2 (H/H) mice, and analyzed by flow cytometry using species-specific anti-CD2 antibodies. Murine CD2 protein was detected in wild-type mice, while human CD2 protein was exclusively detected in B-hCD2 mice.
Protein expression analysis in NK cells
Species-specific CD2 protein expression analysis in humanized B-hCD2 mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hCD2 (H/H) mice, and analyzed by flow cytometry using species-specific anti-CD2 antibodies. Murine CD2 protein was detected in wild-type mice, while human CD2 protein was exclusively detected in B-hCD2 mice.
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Analysis of spleen immune cells
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Analysis of spleen leukocyte subpopulations by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen cell types in spleen. Values are expressed as mean ± SEM.
Analysis of spleen T cell subpopulations by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3+ T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells and Tregs in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen T cell subtypes. Values are expressed as mean ± SEM.
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Analysis of lymph node immune cells
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Analysis of lymph node leukocyte subpopulations by flow cytometry. Leukocytes were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells and NK cells in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these lymph node cell subtypes. Values are expressed as mean ± SEM.
Analysis of lymph node T cell subpopulations by flow cytometry. Leukocytes were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3+ T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells and Tregs in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these lymph node T cell subtypes. Values are expressed as mean ± SEM.
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Analysis of blood immune cells
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Analysis of blood leukocyte subpopulations by flow cytometry. Blood cells were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these blood cell subtypes. Values are expressed as mean ± SEM.
Analysis of blood T cell subpopulations by flow cytometry. Blood cells were isolated from wild-type C57BL/6 and humanized B-hCD2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3+ T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells and Tregs in humanized B-hCD2 mice were similar to those in wild-type mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these blood T cell subtypes. Values are expressed as mean ± SEM.
