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• TGFBR2 humanized mice are genetically engineered mouse models in which the chimeric coding sequence containing the mouse signal peptide, human extracellular domain, and mouse transmembrane/intracellular domains is inserted into exon 2 of mouseTgfbr2 locus, enabling expression of human TGFBR2 in vivo. 
• Transforming growth factor-beta receptor II (TGFBR2) is a critical receptor in the TGF-β signaling pathway involved in immune regulation, cell growth, and tissue homeostasis. Aberrant TGF-β/TGFBR2 signaling is implicated in cancer, fibrosis, and immune diseases, making TGFBR2 a key target in translational research. 
• Phenotypic analyses confirm that human TGFBR2 mRNA and protein are detectable only in homozygous TGFBR2 humanized mice but not in wild-type controls. Leukocyte and T cell subpopulation profiles are comparable to wild-type animals. These findings demonstrate that humanization of Tgfbr2 does not disrupt immune homeostasis and supports functional studies relevant to human physiology. 

Key Advantages

• Physiological Human TGFBR2 Expression: Human TGFBR2 is expressed under native regulatory control, facilitating translational relevance.
• Preserved Immune Homeostasis: Comprehensive flow cytometry shows no significant changes in leukocyte, T cell, B cell, or NK cell distributions compared with wild-type mice. 
• Suitable for Functional Studies: Validated expression enables investigation of human TGF-β pathway function in vivo.
• Translatable for Therapeutic Evaluation: Can be used to assess human-specific effects of TGF-β-targeted therapeutic agents or pathway modulators.
• Stable Genetic Model: Human receptor expression stability supports longitudinal and mechanistic studies.

Validation

• mRNA Expression Validation: RT-PCR analysis shows mouse Tgfbr2 mRNA is detectable only in wild-type splenocytes, whereas human TGFBR2 mRNA is exclusively found in homozygous TGFBR2 humanized mice. 
• Protein Expression Validation: Flow cytometry with a species-specific TGFBR2 antibody confirms exclusive human TGFBR2 detection in homozygous TGFBR2 humanized mice. 
• Immunophenotyping Validation: FACS analyses of spleen, lymph node, and blood leukocytes show that percentages of T cells, B cells, NK cells, monocytes, and macrophages are similar between TGFBR2 humanized mice and C57BL/6 controls, demonstrating normal immune cell development and distribution. 

Application

• Translational Pathway Studies: Ideal for investigating human TGF-β/TGFBR2 signaling in physiological and pathological contexts.
• Functional Evaluation of Therapeutic Candidates: Useful for preclinical assessment of TGF-β pathway modulators, antibodies, or small molecules targeting human TGFBR2.
• Immunology Research: Supports studies in immune regulation and tolerance mechanisms where TGF-β signaling is critical.
• Disease Modeling: Applicable to research in fibrosis, cancer progression, and immune-related disorders associated with TGF-β signaling dysregulation.

The chimeric coding sequence—comprising the mouse signal peptide, human extracellular domain, and mouse transmembrane and intracellular domains—was inserted into exon 2 of the mouse Tgfbr2 locus to generate TGFBR2 humanized mice.

Strain-specific analysis of TGFBR2 gene expression was performed in wild-type (WT) mice (+/+) and homozygous TGFBR2 humanized mice by RT-PCR. Mouse Tgfbr2 mRNA was detectable only in splenocytes of WT mice (+/+). Human TGFBR2 mRNA was detectable exclusively in homozygous TGFBR2 humanized mice but not in WT mice (+/+).

Strain-specific TGFBR2 expression analysis was performed in homozygous TGFBR2 humanized mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous TGFBR2 humanized mice, and analyzed using species-specific TGFBR2 antibodies. As the mouse TGFBR2 antibody cross-reacts with human TGFBR2, mouse TGFBR2 was detectable in both wild-type mice and homozygous TGFBR2 humanized mice. Human TGFBR2 was exclusively detectable in homozygous TGFBR2 humanized mice but not in wild-type mice.

Splenocytes were isolated from female C57BL/6 mice and homozygous TGFBR2 humanized mice (n = 3, 9 weeks old). Flow cytometry analysis was performed to assess leukocyte subpopulations. (A) Representative FACS plots. Single live cells were gated for the CD45⁺ population and used for further analysis. (B) Percentages of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes, and macrophages in homozygous TGFBR2 humanized mice were similar to those in C57BL/6 mice, demonstrating that introduction of human TGFBR2 does not alter leukocyte development, differentiation, or distribution in spleen. Values are expressed as mean ± SEM.

Splenocytes were isolated from female C57BL/6 mice and homozygous TGFBR2 humanized mice (n = 3, 9 weeks old). Flow cytometry was performed to assess T cell subpopulations. (A) Representative FACS plots. Single live CD45⁺ cells were gated for the CD3⁺ T cell population. (B) Percentages of CD8⁺ T cells, CD4⁺ T cells, and regulatory T cells (Tregs) in homozygous TGFBR2 humanized mice were similar to those in C57BL/6 mice, indicating that humanization of TGFBR2 does not affect T cell development or distribution in spleen. Values are expressed as mean ± SEM.

Leukocytes were isolated from lymph nodes of female C57BL/6 mice and homozygous TGFBR2 humanized mice (n = 3, 9 weeks old). Flow cytometry analysis was performed to assess leukocyte subpopulations. (A) Representative FACS plots. Single live cells were gated for the CD45⁺ population. (B) Percentages of T cells, B cells, and NK cells in homozygous TGFBR2 humanized mice were similar to those in C57BL/6 mice, demonstrating that introduction of human TGFBR2 does not alter leukocyte development or distribution in lymph nodes. Values are expressed as mean ± SEM.

Leukocytes were isolated from lymph nodes of female C57BL/6 mice and homozygous TGFBR2 humanized mice (n = 3, 9 weeks old). Flow cytometry was performed to assess T cell subpopulations. (A) Representative FACS plots. Single live CD45⁺ cells were gated for CD3⁺ T cells. (B) Percentages of CD4⁺ T cells, CD8⁺ T cells and Tregs in homozygous TGFBR2 humanized mice were similar to those in C57BL/6 mice, indicating preserved T cell development and distribution in lymph nodes. Values are expressed as mean ± SEM.

Blood cells were isolated from female C57BL/6 mice and homozygous TGFBR2 humanized mice (n = 3, 9 weeks old). Flow cytometry analysis was performed to assess blood T cell subpopulations. (A) Representative FACS plots. Single live CD45⁺ cells were gated for CD3⁺ T cells. (B) Percentages of CD8⁺ T cells, CD4⁺ T cells, and Tregs in homozygous TGFBR2 humanized mice were similar to those in C57BL/6 mice, demonstrating that introduction of human TGFBR2 does not alter T cell development or distribution in peripheral blood. Values are expressed

Strain-specific TGFBR2 expression analysis was performed in homozygous TGFBR2 humanized mice by flow cytometry. Spleens were collected from wild-type mice (+/+) and homozygous TGFBR2 humanized mice and analyzed using an anti-TGFBR2 antibody. Human TGFBR2 was exclusively detectable in homozygous TGFBR2 humanized mice but not in wild-type mice.

Q1: What are TGFBR2 humanized mice?

TGFBR2 humanized mice are genetically engineered mice that express human TGFBR2 receptor in place of the mouse counterpart, enabling translational studies of human TGF-β signaling in vivo.

Q2: How is human TGFBR2 expression confirmed in this model?

Human TGFBR2 expression is validated by RT-PCR and flow cytometry with species-specific antibodies, showing presence only in homozygous TGFBR2 humanized mice and not in wild-type controls. 

Q3: Does humanization of TGFBR2 affect immune cell populations?

No. Flow cytometry analyses indicate that leukocyte and T cell subpopulation distributions in spleen, lymph nodes, and blood are similar between TGFBR2 humanized mice and wild-type mice, indicating preserved immune development. 

Q4: What research applications are suitable for TGFBR2 humanized mice?

This model is suitable for mechanistic exploration of human TGF-β signaling, functional evaluation of human-specific therapeutics, and research in conditions where TGF-β pathways play a central role, such as fibrosis, cancer, and immune diseases.