Basic Information
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Targeting Strategy
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Gene targeting strategy for B-hCD200/hCD200R mice.
Exons 1-5 of mouse Cd200 (encoding the full-length protein) were replaced by human CD200 exons 1-7 in B-hCD200/hCD200R mice.
Exons 2-5 of mouse Cd200r1 (encoding the extracellular domain) were replaced by human CD200R1 exons 2-5 in B-hCD200/hCD200R mice.
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Protein Expression
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Strain specific CD200 expression analysis in homozygous humanized B-hCD200/hCD200R mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD200/hCD200R mice (H/H; H/H), and analyzed by flow cytometry with species-specific anti-CD200 antibody. Mouse CD200 was detectable in WT mice (+/+). Human CD200 was exclusively detectable in homozygous B-hCD200/hCD200R mice (H/H;H/H) but not in WT mice (+/+).
Strain specific CD200R expression analysis in homozygous humanized B-hCD200/hCD200R mice by flow cytometry.
Peritoneal exudative macrophages were collected from wild type (WT) mice (+/+) and homozygous B-hCD200/hCD200R mice (H/H; H/H), and analyzed by flow cytometry with species-specific anti-CD200R antibody. Mouse CD200R was detectable in WT mice (+/+). Human CD200R was exclusively detectable in homozygous B-hCD200/hCD200R mice (H/H; H/H) but not in WT mice (+/+).
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Spleen Leukocyte Profiling
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Analysis of spleen leukocyte subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and humanized B-hCD200/hCD200R mice (n=3, 8 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 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-hCD200/hCD200R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD200/hCD200R 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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Spleen Lymphocyte Profiling
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Analysis of spleen T cell subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hCD200/hCD200R mice (n=3, 8 week-old). Flow cytometry analysis of the splenocytes was performed to assess T cell subsets. 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-hCD200/hCD200R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD200/hCD200R 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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