Basic Information

Strain Name
C57BL/6-Cd3etm1(CD3E)Cd3dtm1(CD3D)Cd3gtm1(CD3G)/Bcgen
Common Name
B-hCD3EDG mice
Background
C57BL/6
Catalog Number
110039
Related Genes
CD3E, CD3D, CD3G

Description

The protein encoded by CD3-epsilon 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. This complex plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. The genes encoding the epsilon, gamma and delta polypeptides are located in the same cluster on chromosome 11. The epsilon polypeptide plays an essential role in T-cell development. T-cell surface glycoprotein CD3 delta chain is a protein that in humans is encoded by the CD3D gene. CD3D has been shown to interact with CD8A. T-cell surface glycoprotein CD3 gamma chain is a protein that in humans is encoded by the CD3G gene, defects in CD3G are associated with T cell immunodeficiency.

Targeting Strategy

Gene targeting strategy for B-hCD3EDG mice. The exons 2-8 of mouse Cd3e gene that  encode the Full-length protein domain were replaced by human CD3E exons 2-9 in B-hCD3EDG mice. The exons 1-5 and exons 1-6 of mouse Cd3d, Cd3g gene that  encode the Full-length protein domain were replaced by human CD3D exons 1-5 and CD3G exons 1-7  respectively in B-hCD3EDG mice.

Details

mRNA expression analysis

Strain specific analysis of CD3EDG gene expression in WT and hCD3EDG mice by RT-PCR. Mouse Cd3edg mRNA was detectable in thymocytes of wild-type (+/+) and heterozygous (H/+) mice. Human CD3EDG mRNA was detectable only in H/+ but not in +/+ mice.

Thymocytes and splenocyte number in B-hCD3EDG mice

(A) Size of thymuses and spleens from C57BL/6 and B-hCD3EDG mice (n=2).  (B) The number of thymocytes in C57BL/6 and  B-hCD3EDG  mice was similar, but splenocyte in B-hCD3EDG mice was higher than that in C57BL/6.

Phenotypic analysis 

Analysis of spleen leukocytes cell subpopulations in B-hCD3EDG mice

A

B

Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hCD3EDG mice (n=3, 5 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, B, NK, DC, Granulocyte, Monocyte, and macrophage cells in heterozygous B-hCD3EDG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3EDG in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen.

 

Analysis of spleen T cell subpopulations in B-hCD3EDG mice

A

B

Analysis of spleen T cell subpopulations by FACS.Splenocytes were isolated from female C57BL/6 and B-hCD3EDG mice (n=3, 5 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, CD4, and Treg cells in heterozygous B-hCD3EDG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3EDG 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.

 

Analysis of thymus leukocytes cell subpopulations in B-hCD3EDG mice

A

B

Analysis of thymus leukocyte subpopulations by FACS. Thymocytes were isolated from female C57BL/6 and B-hCD3EDG mice (n=3, 5 week-old) . Flow cytometry analysis of the thymocytes 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 CD4/CD8  cells, B cells in heterozygous B-hCD3EDG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3EDG in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in thymus.

 

Analysis of  thymus T cell subpopulations in B-hCD3EDG mice

A

B

 

Analysis of thymus T cell subpopulations by FACS. Thymocytes were isolated from female C57BL/6 and B-hCD3EDG mice (n=3, 5 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, CD4, and Treg cells in heterozygous B-hCD3EDG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hCD3EDG in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in thymus. Values are expressed as mean ± SEM.

 

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