B-hIL4/hIL4RA mice(C.B6)_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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Text

B-hIL4/hIL4RA mice(C.B6)

Strain Name	C57BL/6-Il4^{tm2(IL4)Bcgen} Il4ra^{tm1(IL4RA)Bcgen}/Bcgen	Common Name	B-hIL4/hIL4RA mice(C.B6)
Background	BALB/cCrSlcNifdc.C57BL/6JNifdc	Catalog number	111889
Aliases	BCGF-1, BCGF1, BSF-1, BSF1, IL-4 CD124, IL-4RAA, IL4R

Protein expression analysis

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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.

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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.

The number of BALF immune cells in mouse asthma model

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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

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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 show 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

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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). (a) Mucus (b)Eosinophils

In vivo efficacy of anti-human IL4RA antibody with asthma mouse model

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Measurement of enhanced pause (Penh) in mouse asthma model

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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 24h 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)].