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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Strain specific analysis of CD2 gene expression in wild type (WT) mice and B-hCD2 mice by RT-PCR.
Mouse Cd2 mRNA was detectable only in splenocytes of WT mice (+/+). Human CD2 mRNA was detectable only in homozygous B-hCD2 mice (H/H) but not in WT mice (+/+).
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Protein expression analysis in T cells
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Strain specific CD2 expression analysis in homozygous B-hCD2 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD2 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD2 antibody. Mouse CD2 was detectable in WT mice (+/+). Human CD2 was exclusively detectable in homozygous B-hCD2 mice (H/H) but not in WT mice (+/+).
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Protein expression analysis in B cells
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Strain specific CD2 expression analysis in homozygous B-hCD2 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD2 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD2 antibody. Mouse CD2 was detectable in WT mice (+/+). Human CD2 was exclusively detectable in homozygous B-hCD2 mice (H/H) but not in WT mice (+/+).
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Protein expression analysis in NK cells
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Strain specific CD2 expression analysis in homozygous B-hCD2 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD2 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD2 antibody. Mouse CD2 was detectable in WT mice (+/+). Human CD2 was exclusively detectable in homozygous B-hCD2 mice (H/H) but not in WT mice (+/+).
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Analysis of spleen leukocytes cell subpopulations in B-hCD2 mice
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Analysis of spleen leukocyte subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hCD2 mice (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 cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.
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Analysis of spleen T cell subpopulations in B-hCD2 mice
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Analysis of spleen T cell subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hCD2 mice (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+ T cells, CD4+ T cells and Treg cells in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 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.
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Analysis of lymph node leukocytes cell subpopulations in B-hCD2 mice
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Analysis of lymph node leukocyte subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hCD2 mice (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 cells, B cells and NK cells in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.
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Analysis of lymph node T cell subpopulations in B-hCD2 mice
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Analysis of lymph node T cell subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hCD2 mice (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+ T cells, CD4+ T cells and Treg cells in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 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.
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Analysis of blood leukocytes cell subpopulations in B-hCD2 mice
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Analysis of blood leukocyte subpopulations by FACS.
Blood cells were isolated from female C57BL/6 and B-hCD2 mice (n=3, 6 week-old). Flow cytometry analysis of the blood 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 cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.
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Analysis of blood T cell subpopulations in B-hCD2 mice
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Analysis of blood T cell subpopulations by FACS.
Blood cells were isolated from female C57BL/6 and B-hCD2 mice (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+ T cells, CD4+ T cells and Treg cells in homozygous B-hCD2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD2 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.
