hIL2/hIL2RA/hIL2RB/hIL2RG

Fig1. Detection of hIL-2 expression by ELISA in the hIL-2 knockin mice (Wild type and heterozygous hIL-2 KI mice were administrated with concanavalin).

Fig2. Intracellular staining of IL-2 by FACS post stimulation with anti-mCD3/anti-mCD28 (In collaboration with CrownBio).

Fig3. IL-2RA humanized mouse expresses hlL-2RA (hCD25) on naïve Treg in blood (In collaboration with CrownBio).

Fig4. hlL-2RA is highly expressed on activated Tregs in spleen in the IL-2RA huamnzied mouse (In collaboration with CrownBio).

Fig5. hlL-2RA is highly expressed on activated NK cells in spleen in the IL-2RA humanized mouse (In collaboration with CrownBio).

Fig6. Immunotype in hIL2/hIL2RA/hIL2RB/hIL2RG qKI mice.

Fig7. Immunotype in hIL2/hIL2RA/hIL2RB/hIL2RG qKI mice in Blood.

Fig8. Immunotype in hIL2/hIL2RA/hIL2RB/hIL2RG qKI mice in Spleen.

Fig9. Detection of Signaling integrity in hIL2/hIL2RA/hIL2RB/hIL2RG qKI mice.

Note. Splenocytes of C57BL/6 mice were treated with anti-mCD3 and mIL-2(10 ng/ml) for 72h. Splenocytes of hIL2/hIL2RA/hIL2RB/hIL2RG qKI mice were treated with anti-mCD3 and hIL-2(10 ng/ml) for 72h.

Fig.10 Human IL-2 induced the phosphorylation of Stat5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg in the C57BL/6 WT mouse (n=2, 6 weeks old), hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 14 weeks old) and hIL2/hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 17 weeks old). 

Splenocytes were collected from the spleen, treated with different concentrations of human IL-2 for 15 min and analyzed by flow cytometry. As shown in the figure, human IL-2 induced pSTAT5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg.

Fig.11 Mouse IL-2 induced the phosphorylation of Stat5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg in the C57BL/6 WT mouse (n=2, 6 weeks old), hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 14 weeks old) and hIL2/hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 17 weeks old).

Splenocytes were collected from the spleen, treated with different concentrations of mouse IL-2 for 15 min and analyzed by flow cytometry. As shown in the figure, mouse IL-2 induced pSTAT5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg. However, the activation of mouse IL-2 is weaker than that of human IL-2 in hIL2RA/hIL2RB/hIL2RG knockin mice and hIL2/hIL2RA/hIL2RB/hIL2RG knockin mice.

Fig.12 Human IL-15 induced the phosphorylation of Stat5 in CD4+ T cells, CD8+ T cells, NK cells and Treg in the C57BL/6 WT mouse (n=2, 6 weeks old), hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 14 weeks old) and hIL2/hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 17 weeks old). 

Splenocytes were collected from the spleen, treated with different concentrations of human IL-15 for 15 min, and analyzed by flow cytometry. As shown in the figure, human IL-15 induced pSTAT5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg.

Fig.13 Mouse IL-15 induced the phosphorylation of Stat5 in CD4+ T cells, CD8+ T cells, NK cells and Treg in the C57BL/6 WT mouse (n=2, 6 weeks old), hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 14 weeks old) and hIL2/hIL2RA/hIL2RB/hIL2RG knockin mice (n=2, 17 weeks old). 

Splenocytes were collected from the spleen, treated with different concentrations of mouse IL-15 for 15 min, and analyzed by flow cytometry. As shown in the figure, mouse IL-15 induced pSTAT5 in CD4+ T cells, CD8+ T cells, NK cells, and Treg in a dose dependent manner. However, the activation of mouse IL-15 is weaker than that of human IL-15 in all mice.

Fig.14 Analysis of the phosphorylation of Stat5 in CD4+ T cells, CD8+ T cells, and Treg in human PBMC. 

PBMC were treated with different concentrations of human IL-2, human IL-15, mouse IL-2, and mouse IL-15 for 15 min, and analyzed by flow cytometry. As shown in the figure, human IL-2, human IL-15, and mouse IL-2 induced pSTAT5 in CD4+ T cells, CD8+ T cells, and Treg in a dose dependent manner. However, the activation of mouse IL-2 is weaker than that of human IL-2 and human IL-15.