B-hIL4/hIL4RA mice(C.B6)

C.B6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen/Bcgen • 111889

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B-hIL4/hIL4RA mice
B-hIL4/hIL4RA/hIL13 mice

B-hIL4/hIL4RA mice(C.B6)

Product nameB-hIL4/hIL4RA mice(C.B6)
Catalog number111889
Strain nameC.B6-Il4tm2(IL4)Bcgen Il4ratm1(IL4RA)Bcgen/Bcgen
Strain backgroundC57BL/6JNifdc; BALB/cCrSlcNifdc
AliasesBCGF-1, BCGF1, BSF-1, BSF1, IL-4CD124, IL-4RAA, IL4R
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  • Targeting strategy
  • Phenotypic analysis
  • Efficacy

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      Targeting strategy

      Gene targeting strategy for B-hIL4/hIL4RA mice(C.B6). The exons 1-4 of mouse Il4 gene that encode the full length protein were replaced by human IL4 exons 1-4 in B-hIL4/hIL4RA mice. The exons 4-7 of mouse Il4ra gene that encode the extracellular domain were replaced by human IL4RA exons 4-7 in B-hIL4/hIL4RA mice. B-hIL4/hIL4RA mice(C.B6) was obtained by backcrossing B-hIL4/hIL4RA mice with wild-type BALB/c mice for 10 generations.

      Protein expression analysis
      Strain specific IL4 expression analysis in homozygous B-hIL4/hIL4RA mice(C.B6) by ELISA. Serum was collected from wild-type BALB/c mice (+/+; +/+) and homozygous B-hIL4/hIL4RA mice(C.B6) (H/H; H/H) stimulated with anti-CD3ε in vivo, and analyzed by ELISA with species-specific IL4 ELISA kit. Mouse IL4 was detectable in wild-type BALB/c mice. Human IL4 was exclusively detectable in homozygous B-hIL4/hIL4RA mice(C.B6) but not in wild-type BALB/c mice.
      Strain specific IL4RA expression analysis in homozygous B-hIL4/hIL4RA mice(C.B6) by flow cytometry. Splenocytes were collected from wild-type BALB/c mice (+/+; +/+) and homozygous B-hIL4/hIL4RA mice(C.B6) (H/H; H/H), and analyzed by flow cytometry with species-specific anti-IL4RA antibody. Mouse IL4RA was detectable in wild-type BALB/c mice. Human IL4RA was exclusively detectable in homozygous B-hIL4/hIL4RA mice(C.B6) but not in wild-type BALB/c mice.
      In vivo efficacy of anti-human IL4RA antibody with asthma mouse model
      Measurement of enhanced pause (Penh) in mouse asthma model

      Measurement of enhanced pause (Penh) by whole-body plethysmography. Airway responses following the exposure to increasing doses of methacholine (MCh) were measured for each mouse 24 h after the final allergen or PBS exposure using the whole-body plethysmography. The y-axis represents the Penh absolute value. Increasing doses of methacholine were administered by aerosols. B-hIL4/hIL4RA mice(C.B6) (n = 6) exposed to OVA showed a significant increase in airway hyperreactivity to MCh when compared to B-hIL4/hIL4RA mice(C.B6) exposed to PBS inhalation (n = 6) (*P<0.05, *** P<0.01). Penh values were significantly decreased in B-hIL4/hIL4RA mice(C.B6) treated with dupilumab (in house) when compared to B-hIL4/hIL4RA mice(C.B6) treated with hIgG4 (n = 6) after allergen exposure [MCh doses of 50mg/mL (#P<0.05)].

      The number of BALF immune cells in mouse asthma model

      Analysis of immune cells in BALF by FACS. BALF immune cells were isolated from B-hIL4/hIL4RA mice(C.B6) (n=6). The number and proportion of eosinophils were analyzed by flow cytometry under the treatment of PBS/dupilumab (in house). After treatment of dupilumab (in house), the number of CD45+ cells and eosinophils were much lower than the positive control in homozygous B-hIL4/hIL4RA mice(C.B6).

      OVA specific and total IgE production in serum of mouse asthma model

      IgE production in serum of mouse asthma model. Serum was collected at the study endpoint. IgE levels responded to OVA-specific antibody and total IgE levels were analyzed. The results showed that the levels of IgE in mice treated with dupilumab (in house) is much lower than that in untreated mice.

      H&E staining in asthma-like model in B-hIL4/hIL4RA mice

      H&E staining of asthma-like model in B-hIL4/hIL4RA mice(C.B6).Lung tissues were collected at the study endpoint. H&E staining results showed that the lung tissues from B-hIL4/hIL4RA mice(C.B6) exposed to PBS aerosols did not show any inflammation. OVA exposure resulted in a significant increase in peribronchial and perivascular inflammation in B-hIL4/hIL4RA mice(C.B6). A significant reduction in eosinophils infiltration was observed in mice treated with dupilumab (in house).