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B-hCD3E/hCD28 mice
Strain Name C57BL/6-Cd3etm2(CD3E)Cd28tm1(CD28) /Bcgen Common Name  B-hCD3E/hCD28 mice
Background C57BL/6 Catalog number  121248
Related Genes 
CD3E,CD3e molecule, IMD18, T3E, TCRE; CD28, CD28 molecule, Tp44

Gene description


The CD3E molecule, epsilon encoded by CD3E gene is a polypeptide, which together with CD3-gamma, -delta and -zeta, and the T-cell receptor alpha/beta and gamma/delta heterodimers, forms the T cell receptor-CD3 complex. The CD3 complex, a common surface marker on T cells, has important functions not only as an essential component in forming the T cell receptor (TCR)-CD3 complex, but also as an external signal transducer; therefore, the CD3 complex is one of the target molecules to modulate T cell functions. The epsilon polypeptide plays an essential role in T-cell development.
CD28 (CD28 molecule) is one of the proteins expressed on T cells that provide co-stimulatory signals required for T cell activation and survival. T cell stimulation through CD28 in addition to the T-cell receptor (TCR) can provide a potent signal for the production of various interleukins (IL-6 in particular). CD28 is the only B7 receptor constitutively expressed on naïve T cells. Association of the TCR of a naive T cell with MHC:antigen complex without CD28:B7 interaction results in a T cell that is anergic. The agonistic antibody of CD28 is now under phase II clinical trials. 

Protein expression analysis

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Strain specific CD3E expression analysis in homozygous B-hCD3E/hCD28 mice by flow cytometry. 
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3E/hCD28 mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD3E antibody. Mouse CD3E was detectable in WT mice (+/+). Human CD3E was exclusively detectable in homozygous B-hCD3E/hCD28 mice (H/H;H/H) but not in WT mice (+/+).

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Strain specific CD28 expression analysis in homozygous B-hCD3E/hCD28 mice by flow cytometry. 
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3E/hCD28 mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD28 antibody. Mouse CD28 was detectable in WT mice (+/+). Human CD28 was exclusively detectable in homozygous B-hCD3E/hCD28 mice (H/H;H/H) but not in WT mice (+/+).

Analysis of spleen leukocytes cell subpopulations in B-hCD3E/hCD28 mice



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Analysis of spleen leukocyte subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.

Analysis of spleen T cell subpopulations in B-hCD3E/hCD28 mice

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Analysis of spleen T cell subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.

Analysis of thymus T cell subpopulations in B-hCD3E/hCD28 mice

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Analysis of thymus T cell subpopulations by FACS.
Thymocytes were isolated from female C57BL/6 and B-hCD3E/hCD28 mice (n=3, 6 week-old). Flow cytometry analysis of the thymocytes 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in thymus. Values are expressed as mean ± SEM. 

Analysis of lymph node leukocytes cell subpopulations in B-hCD3E/hCD28 mice

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Analysis of lymph node leukocyte subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.

Analysis of lymph node T cell subpopulations in B-hCD3E/hCD28 mice



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Analysis of lymph node T cell subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.

Analysis of blood leukocytes cell subpopulations in B-hCD3E/hCD28 mice

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Analysis of blood leukocyte subpopulations by FACS.
Blood cells were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.

Analysis of blood T cell subpopulations in B-hCD3E/hCD28 mice

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Analysis of blood T cell subpopulations by FACS.
Blood cells were isolated from female C57BL/6 and B-hCD3E/hCD28 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-hCD3E/hCD28 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3E/hCD28 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.