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Strain Name
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C57BL/6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen
Il13tm1(IL13)Bcgen Il13ra1tm1(IL13RA1)Bcgen /Bcgen
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Common Name
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B-hIL4/hIL4RA/hIL13/hIL13RA1 mice
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Background
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C57BL/6
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Catalog number
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112812
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Aliases
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IL4: interleukin 4
IL4RA: interleukin 4 receptor, IL4R
Il13: interleukin 13
IL13RA1: CD213A1, CT19, IL-13Ra, NR4
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Protein expression analysis in serum
Strain specific IL4 expression analysis in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice by ELISA. Serum were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1mice (H/H; H/H; H/H; H/H) stimulated with anti-mCD3Ɛ antibody in vivo, and analyzed by ELISA with species-specific IL4 ELISA kit. Mouse IL4 was detectable in wild-type C57BL/6 mice. Human IL4 was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
Protein expression analysis in spleen
Strain specific IL4RA expression analysis in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice (H/H; H/H; H/H; H/H), and analyzed by flow cytometry with species-specific anti-IL4RA antibody. Mouse IL4RA was detectable in wild-type mice, Human IL4RA was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
Protein expression analysis
Strain specific IL13 expression analysis in homozygous B-hIL4//hIL4RA/hIL13/hIL13RA1 mice by ELISA. CD4+ T cells were collected from wild-type mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice (H/H; H/H; H/H; H/H)，and then induction of CD4 + T cells differentiation into Th2 cells, supernatants from Th2 cells culture was analyzed by ELISA with species-specific IL13 ELISA kit. Mouse IL13 was detectable in wild-type mice. Human IL13 was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
Functional analysis
Assessment of IL-13RA1-mediated responses in a model of IL13 induced airway inflammation. Two doses (10 μg per mouse) of mouse IL13 or human IL13 were administered intratracheally every other day for 4 days to wild-type C57BL/6 mice, homozygous B-hIL4/hIL4RA/hIL13 mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice. 24 hours after the final IL13 challenge, the mice were assessed for BALF chemokine. Mouse IL13 and human IL13 strongly induced CCL11 and CCL24 expression in wild-type mice, homozygous B-hIL4/hIL4RA/hIL13 mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice.
In vivo efficacy of anti-human IL4RA antibody and anti-human IL13 antibody
Analysis of immune cells in BALF. B-hIL4/hIL4R/hIL13/hIL13RA1 mice (female, 10-week-old, n=8) were immunized with hIL13 to induce asthma. Anti-human IL4RA antibody (Dupilumab analog, synthesized in-house) and anti-human IL13 antiboday (Lebrikizumab analog, synthesized in-house) were intraperitoneally injected from Day -1 and Day 5. (A&B) The number of CD45+ cells and eosinophils of BALF in the Dupilumab and Lebrikizumab treated groups decreased significantly compared with the hIL13-induced PBS treated group. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001.
Analysis of cytokines in BALF and mouse total IgE in serum. B-hIL4/hIL4R/hIL13/hIL13RA1 mice (female, 10-week-old, n=8) were immunized with hIL13 to induce asthma. Anti-human IL4RA antibody (Dupilumab analog, synthesized in-house) and anti-human IL13 antiboday (Lebrikizumab analog, synthesized in-house) were intraperitoneally injected from Day -1 and Day 5. The mosue CCL11 and CCL24 of BALF in the Dupilumab and Lebrikizumab treated groups decreased significantly compared with the hIL13-induced PBS treated group. The human IL13 of BALF in the Lebrikizumab treated groups increased compared with the hIL13-induced isotype treated group. Serum was collected at the study endpoint. IgE level was analyzed by ELISA. The results showed that the levels of total IgE in mice treated with Dupilumab and Lebrikizumab showed a significant reduction compared with untreated mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001.
H&E staining of asthma-like model in B-hIL4/hIL4R/hIL13/hIL13RA1 mice . Lung tissues were collected at the study endpoint and analyzed with H&E staining. The results showed that the group of mice treated with Dupilumab and Lebrikizumab (in-house) in inflammatory infiltration and mucus secretion in lung tissue was lower than that in untreated mice, indicating that B-hIL4/hIL4R/hIL13/hIL13RA1 mice provide a powerful preclinical model for in vivo evaluation of anti-human IL4RA antibodies and anti-human IL13 antibodies. Values are expressed as mean ± SEM. Significance was determined by unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001.
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