hPD-1/hGITR/hGITRL - 模型资源 - 基锘威

Design of the hPD-1/hGITR/hGITRL mouse

The triple-humanized PD‑1, GITR and GITRL mouse model (hPD-1/hGITR/hGITRL) was generated by intercrossing hPD-1, hGITR and hGITRL mice.‍

hPD-1 was generated by inserting, within the mouse Pd-1 locus, a chimeric PD-1 with the human extracellular and murine transmembrane and intracellular domains.

hGITR was developed by inserting, within the mouse Gitr locus, a chimeric GITR with the human extracellular and the murine intracellular domain.

hGITRL was generated by inserting, within the mouse Gitrl locus, a chimeric GITRL with the human extracellular and the murine intracellular domain.

hPD-1, hGITR and hGITRL expressions are regulated by endogenous mouse promoters.

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hPD-1/hGITR/hGITRL features

PD-1, GITR and GITRL extracellular domains are entirely humanized
hPD-1, hGITR and hGITRL expression driven by the respective endogenous mouse promoter
Fully functional mouse immune system
Lack of expression of the murine target genes, thus avoiding cross-reactivity

hPD-1 is expressed on T cells

Freshly isolated splenocytes from wild-type or PD-1/GITR/GITRL triple-humanized mice. mPD-1 and hPD-1 expression were analyzed on single live CD3⁺, CD4⁺CD25^-cells (conv CD4) and CD4⁺CD25⁺Foxp3⁺ cells (Treg). T cells from lymphocytes incubated in the presence of anti-CD3, anti-CD28 and recombinant murine IL-2.
hGITR is expressed on T cells

Freshly isolated splenocytes from wild-type or PD-1/GITR/GITRL triple humanized mice. mGITR and hGITR expression were analyzed on single live CD3⁺, CD4⁺CD25^-cells (conv CD4) and CD4⁺CD25⁺Foxp3⁺ cells (Treg). Cells from lymphocytes were incubated in the presence of anti-CD3, anti-CD28 and recombinant murine IL-2.
In vivo anti-tumor activity is observed in response to anti-PD-1 treatment in hPD-1 mice

Survival of mice of indicated genotype implanted intracranially with GL261 murine glioma cells and treated with αhPD-1 (pembrolizumab), αmPD-1 or isotype (Mantel-Cox test: ***p<0.001).
hGITR-Fc significantly inhibits ear swelling at 24 and 48 hours post DNFB challenge in hGITR/hGITRL mice

Mice were injected with vehicle, IgG, GITR-Fc or mCTLA4-Ig on day -1, sensitized with DNFB on day 0, and then challenged on the ear on day 5. The mean ear-swelling response was determined at 24 and 48 hours post DNFB challenge.