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B-hTPBG mice
Strain Name C57BL/6-Tpbgtm1(TPBG)/Bcgen Common Name  B-hTPBG mice
Background C57BL/6 Catalog number  111048
Related Genes 
TPBG (Trophoblast glycoprotein), 5T4

Gene description

Trophoblast glycoprotein, also known as TPBG, 5T4, Wnt-Activated Inhibitory Factor 1 or WAIF1, is an antagonist of Wnt/β-catenin signaling pathway. TPBG is expressed in a number of carcinomas, and found in tumors including the colorectal, ovarian, and gastric. Its expression is used as a prognostic aid in these cases. It has very limited expression in normal tissue but is widespread in malignant tumors throughout their development. One study found that 5T4 was present in 85% of a cohort of 72 colorectal carcinomas and in 81% of a cohort of 27 gastric carcinomas. Its confined expression appears to give TPBG the potential to be a target for T cells in cancer immunotherapy. There has been extensive research into its role in antibody-directed immunotherapy through the use of the high-affinity murine monoclonal antibody, mAb5T4, to deliver response modifiers (such as staphylococcus aureus super-antigen) accurately to a tumor. TPBG is also the target of the cancer vaccine TroVax which is in clinical trials for the treatment of a range of different solid tumor types.


mRNA expression analysis


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Strain specific analysis of TPBG gene expression in WT and B-hTPBG mice by RT-PCR. Mouse Tpbg mRNA was detectable in splenocytes of wild-type (+/+) mice. Human TPBG mRNA was detectable only in H/H, but not in +/+ mice. 


Protein expression analysis

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Strain specific analysis of TPBG expression in WT and B-hTPBG mice by western blot. Spine were collected from WT mice and homozygous B-hTPBG (H/H) mice. Mouse TPBG was detectable in WT mice, the observed band size 63 kDa is the target band. Human TPBG was exclusively detectable in homozygous B-hTPBG and hTPBG overexpressed MC38 cells but not WT mice.


Analysis of leukocytes subpopulation in B-hTPBG mice


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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. The same results were observed in the spleen and lymph node (data was not shown).Values are expressed as mean ± SEM.


Analysis of blood T cell subpopulations in B-hTPBG mice

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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. The same results were observed in the spleen and lymph node (data was not shown). Values are expressed as mean ± SEM.


Blood routine test in B-hTPBG mice


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Complete blood count (CBC). Blood from female C57BL/6 and B-hTPBG mice (n=6, 6-8 week-old) was collected and analyzed for CBC. Results indicate that introduction of hTPBG in place of its mouse counterpart does not change blood cell composition and morphology. Values are expressed as mean ± SEM.


Blood chemistry of B-hTPBG mice
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Blood chemistry tests of B-hTPBG mice. Serum from the C57BL/6 and B-hTPBG mice (n=6, 6-8 week-old) was collected and analyzed. There was no differences on either measurement between C57BL/6 and B-hTPBG mice, indicating that introduction of hTPBG in place of its mouse counterpart does not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.