Basic Information
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Targeting strategy
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Gene targeting strategy for B-hOSMR mice. The chimerism CDS composed with human OSMR extracellular domain and mouse Osmr intracellular domain was insert into mouse Osmr gene exon 1 in B-hOSMR mice.
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mRNA and protein expression analysis
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Strain specific analysis of OSMR gene expression in wild-type mice and B-hOSMR mice by RT-PCR and western blot. (A) mRNA expression. Mouse Osmr mRNA was exclusively detectable in the kidney of wild-type mice (+/+). Human OSMR mRNA was exclusively detectable only in homozygous B-hOSMR mice (H/H). (B) Protein expression. Kidney was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hOSMR mice (H/H), and analyzed by western blot with anti-OSMR antibody. The OSMR protein can be detected in homozygous B-hOSMR mice.
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Analysis of leukocytes cell subpopulation in spleen
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Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hOSMR 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 the 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-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that OSMR humanized 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 T cell subpopulation in spleen
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Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hOSMR 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 TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOSMR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.
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Analysis of leukocytes cell subpopulation in lymph node
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Analysis of lymph node leukocyte subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hOSMR mice (n=3, 6-week-old). Flow cytometry analysis of the lymph node 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, B cells, and NK cells in homozygous B-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that OSMR humanized 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 T cell subpopulation in lymph node
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Analysis of lymph node T cell subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hOSMR mice (n=3, 6-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOSMR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.
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Analysis of leukocytes cell subpopulation in blood
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Analysis of blood leukocyte subpopulations by FACS. Blood were isolated from female C57BL/6 and B-hOSMR mice (n=3, 6-week-old). Flow cytometry analysis of the blood 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, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that OSMR humanized 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 T cell subpopulation in blood
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Analysis of blood T cell subpopulations by FACS. Blood were isolated from female C57BL/6 and B-hOSMR mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hOSMR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOSMR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.
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Blood routine test in B-hOSMR mice
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Complete blood count (CBC). Blood from female C57BL/6 and B-hOSMR mice (n=6, 7 week-old) was collected and analyzed for CBC. The red blood cells, hemoglobin, and platelet count were decreased in B-hOSMR mice. Values are expressed as mean ± SEM.
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Summary
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mRNA and protein expression analysis
Human OSMR mRNA was exclusively detectable only in homozygous B-hOSMR mice (H/H). The OSMR protein can be detected in homozygous B-hOSMR mice.
Leukocytes cell subpopulation analysis：
OSMR humanized does not change the overall development, differentiation or distribution of immune cell types in spleen, lymph node and blood.
Complete blood count (CBC).
The red blood cells, hemoglobin, and platelet count were decreased in B-hOSMR mice.
