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The humanized GITR/Foxp3 model was generated by intercrossing hGITR and Foxp3-IRES-mRFP (FIR) reporter mice. The FIR's bicistronic reporter expressing a red fluorescent protein (RFP) has been knocked into the endogenous Foxp3 locus and faithfully mirrors Foxp3 expression.

 

Strain Background: C57BL/6J×C57BL/6N

 

Strain Features:

  •   The GITR extracellular domain is entirely humanized
  •   Physiological regulation and expression pattern of the human GITR
  •   Fully functional mouse immune system
  •   Lack of expression of the murine GITR, thus avoiding cross-reactivity
  •   RFP reporter faithfully mirrors Foxp3 expression
  •   The expression of endogenous Foxp3 is not affected by the inserted gene

 

Model Validation:

Figure 1. hGITR expression pattern in hGITR/Foxp3 mice recapitulates mGITR expression in wild-type mice.
Expression of human and mouse GITR on αCD3/αCD28-stimulated splenocytes, analyzed by flow cytometry on Tregs and conventional CD4+ (CD4conv) T cells.

Figure 2. RFP expression mirrors Foxp3 protein expression. Expression of mRFP on non-permeabilized (A) or permeabilized (B) freshly isolated splenocytes (CD3+CD4+) from hGITR/Foxp3 mice. mRFP+ cells were sorted and permeabilized to allow Foxp3 detection (C).

Figure 3. Suppressive function of Treg is preserved. Isolated CD4+CD25- Teff cells were labeled with CTV and cultured with various concentrations of isolated RFP+ Treg cells in presence of antigen-presenting cells treated with mitomycin and αCD3 for 4 days. Proliferation was assessed by flow cytometry (CTV+ cells) on viable cells.

Figure 4. In humanized GITR (hGITR) mice, treatment with hGITR monoclonal antibody can exert an anti-tumor effect. Homozygous hGITR mice were implanted with MB49 bladder cancer cells, and tumor growth was inhibited after the treatment of anti-hGITR.

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