Basic Information
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Targeting strategy
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Gene targeting strategy for B-hIL31/hIL31RA/hOSMR mice.
The exons 1-3 of mouse Il31 gene that encode the mature protein were replaced by human IL31 exons 1-3 in B-hIL31/hIL31RA/hOSMR mice.
The chimeric IL31RA CDS was inserted in exon 4 of mouse Il31ra gene in B-hIL31/hIL31RA/hOSMR mice.
The coding sequences of human OSMR extracellular region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL31/hIL31RA/hOSMR mice.
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mRNA expression analysis (IL31/IL31RA)
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Strain specific analysis of IL31 and IL31RA gene expression in wild-type mice and homozygous B-hIL31/hIL31RA mice by RT-PCR. Testis was collect from wild type C57BL/6 mice (+/+) and B-hIL31/hIL31RA mice (H/H). Mouse Il31 and Il31ra mRNA were exclusively detectable in wild-type mice. Human IL31 and IL31RA mRNA were exclusively detectable in homozygous B-hIL31/hIL31RA mice, but not in wild type mice.
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mRNA and protein expression analysis (OSMR)
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Strain specific analysis of OSMR gene expression in wild-type mice and B-hOSMR mice by RT-PCR and western blot. (A) mRNA expression. Mouse Osmr mRNA was exclusively detectable in the kidney of wild-type mice (+/+). Human OSMR mRNA was exclusively detectable in homozygous B-hOSMR mice (H/H). (B) Protein expression. Kidney was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hOSMR mice (H/H), and analyzed by western blot with anti-OSMR antibody. The OSMR protein can be detected in homozygous B-hOSMR mice.
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hIL31 induces pruritus in B-hIL31/hIL31RA/hOSMR mice
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hIL31 induces pruritus in B-hIL31/hIL31RA/hOSMR mice. hIL-31 are administered to homozygous B-hIL31/hIL31RA/hOSMR mice (female, n=3) on Day0, Day3, Day7, Day10 and record scratching counts for 3h after hIL-31 injection. Record scratching counts before hIL-31 injection as baseline. As shown in the panel, hIL-31 induced pruritus successfully in B-hIL31/hIL31RA/hOSMR mice. An antibody targeting IL-31 signaling was used as positive control, which had shown a significant antipruritic effect in our model (Validated by client).
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Anti-hIL31RA antibodies attenuate scratching behaviors in MC903-induced atopic dermatitis model
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Anti-hIL31RA antibodies attenuate the scratching behaviors in the MC903-induced murine model for atopic dermatitis (AD).
MC903 (calcipotriol) was topically applied to the B-hIL31/hIL31RA/hOSMR mice (female, 10-week-old, 3 mice/group) ear every 2 days for 16 days totally. Meanwhile, the mice were treated with nemolizumab (in-house) or isotype antibodies. Mice in vehicle group were only treated with absolute ethanol. On day 7 and day 17, scratching behavior was recorded. Scratching behaviors on day 7 (A) and day 17 (B). As shown, MC903-treated mice show significantly increased scratching behaviors compared to vehicle treated mice. In the nemolizumab treatment group, blockade of IL-31 signaling with anti-hIL-31RA antibodies significantly attenuates scratching behavior. Data was shown as Mean±SEM, and analyzed using unpaired t-test compared with Isotype group. (*p<0.05).
