B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice

C57BL/6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen Il13tm1(IL13)Bcgen Il13ra1tm1(IL13RA1)BcgenIl31tm1(IL31)Bcgen Il31ratm1(IL31RA)Bcgen Osmrtm1(OSMR)Bcgen/Bcgen • 113830

B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice

Product nameB-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice
Catalog number113830
Strain nameC57BL/6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen Il13tm1(IL13)Bcgen Il13ra1tm1(IL13RA1)BcgenIl31tm1(IL31)Bcgen Il31ratm1(IL31RA)Bcgen Osmrtm1(OSMR)Bcgen/Bcgen
Strain backgroundC57BL/6
NCBI gene ID (Human)
AliasesBSF1; IL-4; BCGF1; BSF-1; BCGF-1; CD124; IL4RA; IL-4RA; P600; IL-13; NR4; CT19; CD213A1; IL-13Ra; IL-31; CRL; GPL; CRL3; GLMR; GLM-R; PLCA2; hGLM-R; IL-31RA; PRO21384; zcytoR17; OSMRB; PLCA1; IL-31RB; OSMRbeta; IL-31R-beta

在此页面上

  • Description
  • Targeting strategy
  • Phenotypic analysis

海报

查看全部

    发表文章

      Description
      • IL‑4 and IL‑13 are shared IL‑4 receptor subunit alpha (IL‑4Rα), as a result, have some overlapping functions. The type I receptor, comprising IL‑4Rα and common cytokine receptor γ-chain (γc), only binds IL‑4. The type II receptor complex, comprising IL‑4Rα and IL‑13Rα1, is the primary receptor for IL‑13 but also binds IL‑4.
      • IL-31 signals via a heterodimeric receptor complex composed of a IL‑31 RA and OSMR, and IL-31 directly binds to IL31RA, not OSMR.
      • The mouse Il4 gene that encodes the full length coding sequence was replaced by human IL4 counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The mouse Il4ra gene that encodes the extracellular region coding sequences was replaced by human IL4RA counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The mouse Il13 gene that encodes the full length coding sequence was replaced by human IL13 counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The mouse Il13ra1 gene that encodes the extracellular protein was replaced by human IL13RA1 counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The mouse Il31 gene that encodes the full-length protein was replaced by human IL31 counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The coding sequences including human IL31RA extracellular region, mouse Il31ra transmembrane and region mouse Il31ra intracellular region were inserted into mouse Il31ra gene in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The coding sequences including human OSMR extracellular region, mouse Osmr transmembrane region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • IL4, IL13, IL4RA and OSMR protein was detectable in wild-type C57BL/6JNifdc mice and B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • Mouse Il4, Il13, Il31, Il31ra and Osmr mRNA were only detectable in wild-type C57BL/6JNifdc mice. Human IL4, IL13, IL31, IL31RA and OSMR mRNA were detectable only in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • Application: This product is used for pharmacodynamics and safety evalsuation of  drugs.
      Targeting strategy

      Gene targeting strategy for B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR 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/hIL13RA1/hIL31/hIL31RA/hOSMR 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/hIL13RA1/hIL31/hIL31RA/hOSMR 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/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The exons 2-9 of mouse Il13ra1 gene that encode the extracellular protein were replaced by human IL13RA1 exons 2-9 in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The mouse Il31 gene that encode the full-length protein was replaced by human IL31 counterpart gene sequences in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The coding sequences including human IL31RA extracellular region, mouse Il31ra transmembrane and region mouse Il31ra intracellular region were inserted into mouse Il31ra gene in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.
      • The coding sequences including human OSMR extracellular region, mouse Osmr transmembrane region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Note: The mice was obtained by B-hIL4/hIL4RA/hIL13/hIL13RA1 mice cross breeding with B-hIL31/hIL31RA/hOSMR mice .

      mRNA expression analysis

      Strain specific analysis of IL4, IL13 and IL31 mRNA expression in wild-type C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by RT-PCR.  Spleen, testis and thymus RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice (H/H;HH;H/H;H/H;H/H;H/H;H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human IL4, IL13 and IL31 primers. Mouse Il4. Il13 and Il31 mRNA were only detectable in wild-type mice. Human IL4, IL13, and IL31 mRNA were exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice  but not in wild-type mice. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Strain specific analysis of IL13RA1, IL31RA, OSMR mRNA expression in C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by RT-PCR. Spleen and testis RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice (H/H;HH;H/H;H/H;H/H;H/H;H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human IL13RA1, IL31RA, OSMR primers. Mouse Il13ra1, Il31ra, Osmr mRNA was only detectable in wild-type mice. Human IL13RA1, IL31RA, OSMR mRNA was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice but not in wild-type mice. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Protein expression analysis

      Strain specific OSMR expression analysis in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by western blot. Brain, liver, lung, kidney and bladder were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice (H/H; H/H; H/H ; H/H; H/H ; H/H; H/H) and analyzed by western blot with anti-OSMR antibody(Proteintech, 10982-1-AP). m/hOSMR was detectable in brain, liver, lung, kidney and bladder of wild-type C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Protein expression analysis in serum

      Strain specific IL-4 expression analysis in wild-type C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by ELISA. Serum was collected from wild-type C57BL/6JNifdc mice (male, n=3, 12-week-old) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice (male, n=3, 12-week-old) stimulated with anti-mCD3ε (BioXCell, BE0001-1) in vivo for 2 hrs. Expression level of mouse IL-4 and human IL-4 were analyzed by ELISA (Mouse IL-4 ELISA Kit : Biolegend, 431104; Human IL-4 ELISA Kit: Biolegend, 430304) . Expression of mouse IL-4 and human IL-4 exhibit normal expression with no cross-react. Values are expressed as mean ± SEM. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Protein expression analysis in supernatant

      Strain specific IL-13 expression analysis in wild-type C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by ELISA. Naive CD4+ T cells were collected from C57BL/6JNifdc mice (+/+) (male, n=3, 12-week-old) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice (male, n=3, 12-week-old) , and then induction of naive CD4+ T cells differentiation into Th2 cells, supernatant from Th2 cells culture was analyzed by ELISA with species-specific IL13 ELISA kit (IL-13 Mouse SimpleStep ELISA Kit : Abcam, ab219634; IL-13 Human SimpleStep ELISA Kit : Abcam, ab288591). Mouse IL-13 was detectable in wild-type mice. Human IL-13 was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice but not in wild-type mice. Values are expressed as mean ± SEM. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Protein expression analysis in spleen

      Strain specific IL4RA expression analysis in wild-type C57BL/6JNifdc mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice  (H/H) stimulated with anti-CD3ε (BioXcell, BE0001)  in vivo for 24 hrs, and protein expression was analyzed with anti-mouse IL4RA antibody (Biolegend, 144804)  and anti-human IL4RA antibody (Biolegend, 355006) by flow cytometry. mIL4RA was detectable in wild-type C57BL/6JNifdc mice. hIL4RA was detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice, but not in wild-type mice. WT: wild-type C57BL/6JNifdc mice; HO: homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice.

      Function analysis-IL4 signaling pathway

      Function analysis of hIL4 signaling pathway in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice. Mice in each group were treated with anti-hIL4RA antibody Dupilumab (in house). (A) Functional experiment scheme diagram. (B) Body weight changes during the treatment. (C) Statistical analysis of ear thickness in each group. (D) Total IgE levels in serum. Serum was collected on day 26 and total IgE levels were measured by ELISA. (n = 6). (E) Hematoxylin and eosin (H&E) staining. (F) Thickness of ear epidermal skin. (G) Score of eosinophils infiltrated in ear epidermal skin. (H) Total score of ear epidermal skin. Ear thickness, IgE levels and infiltration scores of eosinophils in ear skin of the groups treated with Dupilumab were decreased significantly compared to that in the control G2, demonstrating that the B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice provide a powerful preclinical model for in vivo evalsuation of anti-human IL4RA antibodies.

      Function analysis-IL31 signaling pathway

      Function analysis of hIL31 signaling pathway in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice. Mice in G2 were treated with human IL31 protein (hIL31) (in house). (A) Functional experiment scheme diagram. (B) Mouse IL-6 levels in serum. (C) Mouse CCL2 levels in serum. After 1.5 hours of intravenous injection of hIL31, the serum was collected and the levels of IL6 and CCL2 in the serum were measured by ELISA (n = 6). Mouse IL-6 and CCL2 levels in serum were increased after hIL31 stimulation, demonstrating that the IL-31 signaling pathway in B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice is intact.

      * When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hIL4/hIL4RA/hIL13/hIL13RA1/hIL31/hIL31RA/hOSMR mice] (Cat# 113830) was purchased from Biocytogen.
      站点地图