Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD16B mice. The human gene coding sequence was inserted to mouse Hipp11(H11) allele in B-hCD16B mice.
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Protein expression analysis in blood and spleen granulocytes
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Strain specific CD16B expression analysis in homozygous B-hCD16B mice by flow cytometry. Blood (A) and spleen (B) were collected from wild-type mice (+/+) and homozygous B-hCD16B mice (H/H), and analyzed by flow cytometry with species-specific anti-CD16 antibody, respectively. Human CD16B was exclusively detectable in granulocytes of homozygous B-hCD16B mice. Note: the clone of anti-human CD16B antibody is 3G8.
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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 homozygous B-hCD16B 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, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hCD16B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD16B 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 homozygous B-hCD16B 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-hCD16B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD16B 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 blood
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Analysis of blood leukocyte subpopulations by FACS. Blood cells were isolated from female C57BL/6 and homozygous B-hCD16B mice (n=3, 7-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-hCD16B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD16B 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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Summary
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Protein expression analysis:
Human CD16B was exclusively detectable in homozygous B-hCD16B mice but not wild-type mice.Leukocytes profile:
Percentages of T, B, NK cells, monocyte/macrophages, and DC were similar in homozygous B-hCD16B mice and wild-type mice, demonstrating that introduction of hCD16B does not change the overall development, differentiation, or distribution of these cell types in spleen, lymph node leukocytes and blood.
