hGITR/hFoxp3 - 模型资源 - 基锘威

Design of the hGITR/Foxp3 mouse

The humanized GITR/Foxp3 model was generated by intercrossing hGITR and Foxp3-IRES-mRFP (FIR) reporter mice.

The hGITR is developed by Knockin at the mouse GITR locus, and expresses a chimeric GITR with a human extracellular and murine intracellular domain. hGITR is regulated by the endogenous mouse promoter. The FIR's bicistronic reporter expressing a red fluorescent protein (RFP) has been knocked into the endogenous Foxp3 locus and faithfully mirrors Foxp3 expression.

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hGITR/Foxp3 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
Physiological expression of murine Foxp3
RFP reporter faithfully mirrors Foxp3 expression

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 (5 days), analyzed by flow cytometry on Tregs and conventional CD4⁺ (CD4conv) T cells.
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).
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. Results were analysed by Student t test *p<0.001.