Basic Information

Strain Name
C57BL/6-Tpbgtm1(TPBG)/Bcgen
Common Name
B-hTPBG mice
Catalog Number
111048
Background
C57BL/6N
Related Gene
TPBG (Trophoblast glycoprotein), 5T4
NCBI Gene ID

Gene targeting strategy

Gene targeting strategy for B-hTPBG mice. The exon 2 of mouse Tpbg gene that encodes the full-length protein was replaced by human TPBG exon 2 in B-hTPBG mice.

mRNA expression analysis

Species-specific TPBG gene expression analysis in wild-type and humanized B-hTPBG mice by RT-PCR. Murine Tpbg mRNA was detected in splenocytes isolated from wild-type (+/+), while human TPBG mRNA was exclusively detected in homozygous B-hTPBG (H/H) mice.

Protein expression analysis

Species-specific TPBG protein expression analysis in wild-type and humanized B-hTPBG mice. Spinal tissue was isolated from wild-type and homozygous B-hTPBG (H/H) mice and analyzed by western blot using species-specific anti-TPBG antibodies. Murine TPBG protein was detected in wild-type mice (target band size is 63 kDa), while human TPBG protein was exclusively in B-hTPBG mice and hTPBG MC38 cells.

Immune cell profiling in humanized B-hTPBG mice

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

Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hTPBG 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 CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hTPBG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTPBG 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 leukocytes subpopulation in TPBG mice

Analysis of lymph node leukocyte subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hTPBG 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, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hTPBG mice were similar to those in the C57BL/6 mice, demonstrating that TPBG humanized 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-hTPBG mice

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

Analysis of leukocytes subpopulation in B-hTPBG mice

Analysis of blood leukocyte subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hTPBG mice (n=3, 6-week-old). Flow cytometry analysis of the blood cells 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-hTPBG mice were similar to those in the C57BL/6 mice, demonstrating that TPBG humanized 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-hTPBG mice

Analysis of blood T cell subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hTPBG mice (n=3, 6-week-old). Flow cytometry analysis of the blood cells 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-hTPBG mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTPBG 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.