C57BL/6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen Il13tm1(IL13)Bcgen Il13ra1tm1(IL13RA1)Bcgen Tslptm1(TSLP)Bcgen Crlf2tm2(CRLF2)Bcgen/Bcgen • 113627
| Product name | B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice |
|---|---|
| Catalog number | 113627 |
| Strain name | C57BL/6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen Il13tm1(IL13)Bcgen Il13ra1tm1(IL13RA1)Bcgen Tslptm1(TSLP)Bcgen Crlf2tm2(CRLF2)Bcgen/Bcgen |
| Strain background | C57BL/6 |
| NCBI gene ID | 3565,3566,3596,85480,64109 (Human) |
| Aliases | BSF1; IL-4; BCGF1; BSF-1; BCGF-1; CD124; IL4RA; IL-4RA; P600; IL-13; CRL2; TSLPR; CRLF2Y |
Gene targeting strategy for B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice
The exons 1-4 of mouse Il4 gene that encode the full length coding sequence were replaced by human IL4 exons 1-4 in B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice.
The exons 4-7 of mouse Il4ra gene that encode the extracellular region coding sequences were replaced by human IL4RA exons 4-7 in B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice.
The exons 1-4 of mouse Il13 gene that encode the full length coding sequence were replaced by human IL13 exons 1-4 in B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice.
The exons 1-5 of mouse Tslp gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained.
The human TSLP expression is driven by endogenous mouse Tslp promoter, while mouse Tslp gene transcription and translation will be disrupted; A chimeric CDS that encodes human TSLPR extracellular domain, transmembrane and mouse TSLPR cytoplasmic domain, followed by mouse 3’UTR-STOP is inserted right after mouse Tslpr signal peptide sequence to replace part of the exon 2 of mouse Tslpr gene. The chimeric TSLPR protein expression will be driven by endogenous mouse Tslpr promoter, while mouse Tslpr gene transcription and translation will be disrupted.
B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice was developed by cross-mating the B-hTSLP/hTSLPR mice and the B-hIL4/hIL4RA/hIL13 mice together.
Analysis of immune cells in BALF (Bronchoalveolar fluid) by flow cytometry. B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice (male, 15-week-old, n=5) were immunized with OVA etc. to induce asthma. Anti-human TSLP antibody (Tezepelumab analog, synthesized in-house) and anti-human IL13 antibody (IL13-lebrikizumab-hIgG4 analog, synthesized in-house) were intraperitoneally injected to B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice. The results showed that the number of CD45+ cells and eosinophils of BALF in mice treated with anti-human TSLP antibody and anti-human IL13 antibody or a combination of anti-human TSLP antibody and anti-human IL13 antibody decreased significantly compared with the OVA etc.-induced untreated group (G2). Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Mouse total IgE in serum were reduced in the mouse asthma model treated with anti-human TSLP antibody and anti-human IL13 antibody. Serum was collected at the study endpoint. IgE level was analyzed by ELISA. The results showed that the level of total IgE in mice treated with anti-human TSLP antibody and anti-human IL13 antibody or a combination of both were lower than that in untreated mice. Values are expressed as mean ± SEM. Significance was determined by unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001.
H&E staining of asthma-like model in B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice. Lung tissues were collected at the study endpoint and analyzed with H&E staining. The results showed that compared to the untreated group (G2), the group of mice treated with antibodies showed a significant reduction in inflammatory infiltration and mucus secretion in lung tissue, indicating that B-hIL4/hIL4RA/hIL13/hTSLP/hTSLPR mice provide a powerful preclinical model for in vivo evaluation of anti-human TSLP antibody and anti-human IL13 antibody or a combination of both. Values are expressed as mean ± SEM. Significance was determined by unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001.
