• IL4 and IL13 are key type 2 cytokines involved in allergic inflammation, asthma, atopic dermatitis, IgE production, eosinophil recruitment, and epithelial immune responses.
• IL4RA and IL13RA1 form receptor components for IL4/IL13 signaling, making the IL4RA-IL13RA1 axis a major therapeutic pathway in Th2-driven inflammatory disease.
• OX40 and OX40L are costimulatory immune-checkpoint molecules in the TNF/TNFR superfamily that regulate T cell activation, antigen-presenting cell interaction, and inflammatory immune responses.
• B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice combine humanized IL4, IL4RA, IL13, IL13RA1, OX40, and OX40L targets in a C57BL/6 background.
• This model supports in vivo efficacy evaluation of anti-human IL4RA, anti-human IL13, and anti-human OX40L antibodies in atopic dermatitis and asthma-like disease models.

Key Advantages

• Multi-gene humanized IL4/IL4RA/IL13/IL13RA1/OX40/OX40L model for Th2 inflammation research
• Supports anti-human IL4RA, IL13 and OX40L, and related antibody therapies
• Enables atopic dermatitis model readouts including ear thickness, body weight, total IgE, and H&E pathology
• Enables OVA-induced asthma model readouts including BALF immune-cell profiling, serum total IgE, and lung histopathology
• Useful for monotherapy and combination therapy evaluation in allergy, asthma, and atopic dermatitis
• Provides a translational platform for immunology, inflammation, and autoimmune drug development

Validation

• Targeting Strategy Validation: Human IL4, IL4RA, IL13, IL13RA1, OX40, and OX40L sequences were introduced into the corresponding mouse loci to generate a multi-target humanized model.
• AD Model Efficacy Validation: Amlitelimab analog and dupilumab analog reduced ear thickness, total IgE, epidermal thickness, and ear skin pathological score in B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice.
• Asthma Model Efficacy Validation: Anti-human IL4RA, anti-human IL13, and anti-human OX40L antibodies reduced BALF CD45+ cells, eosinophils, serum total IgE, lung inflammatory infiltration, and mucus secretion.
• Combination Therapy Validation: Antibody combinations targeting IL4RA, IL13, and OX40L showed therapeutic activity in OVA-induced asthma-like inflammation.

Application

• Anti-human IL4RA antibody efficacy evaluation
• Anti-human IL13 antibody and IL13 pathway studies
• Anti-human OX40L antibody efficacy evaluation
•  Atopic dermatitis model studies
• OVA-induced asthma and allergic airway inflammation studies
• Combination therapy research for Th2-driven inflammatory disease

In B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice, mouse Il4 exons 1-4, Il13 exons 1-4, Il13ra1 exons 2-9, Ox40 exons 1-5, and Ox40l exons 2-3 were replaced by corresponding human sequences. Mouse Il4ra exons 4-7 encoding extracellular-region coding sequences were replaced by human IL4RA exons 4-7.
This targeting strategy introduces human IL4, IL4RA, IL13, IL13RA1, OX40, and OX40L targets while maintaining the mouse genomic context. B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice were obtained by breeding B-hIL4/hIL4RA/hIL13/hIL13RA1 mice with B-hOX40/hOX40L mice.

Efficacy of anti-human OX40L antibody amlitelimab analog and anti-human IL4RA antibody dupilumab analog in B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice. 
(A&B) Ear thickness and body weight changes during the treatment. (C&D) Total IgE levels in serum. Compared to the untreated group (G2), the treated group with amlitelimab analog (provided by a client) or dupilumab analog (provided by a client) showed a significant reduction in ear thickness. Serum was collected at the study endpoint. IgE level was analyzed by ELISA. The results showed that the levels of total IgE in mice treated with amlitelimab analog or dupilumab analog was lower than that in untreated mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001.

H&E staining of AD model in B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice. 
Ear 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 amlitelimab analog or dupilumab analog showed a significant reduction in epidermal thickness and pathological score of ear skin. 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. AD: Atopic dermatitis.

Analysis of immune cells in BALF (Bronchoalveolar fluid) by flow cytometry. B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice (female, 15-week-old, n=5) were immunized with OVA to induce asthma. Anti-human IL4RA antibody (IL4RA-dupliumab-hIgG4 analog, synthesized in-house), anti-human IL13 antibody (IL13-lebrikizumab-hIgG4 analog, synthesized in-house) and anti-human OX40L antibody  (OX40L-amlitelimab-hIgG4 analog, synthesized in-house) were intraperitoneally injected. BALF was collected at the end of the experiment to detect inflammatory cell infiltration in lung tissue. The results showed that the number of CD45+ cells and eosinophils of BALF in the antibodies treated group decreased significantly compared with the OVA-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 IL4RA antibody, anti-human IL13 antibody and anti-human OX40L 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 IL4RA antibody, anti-human IL13 antibody and anti-human OX40L 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/hIL13RA1/hOX40/hOX40L 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. Values are expressed as mean ± SEM. Significance was determined by unpaired t-test.  *P < 0.05, **P < 0.01, ***P < 0.001.

Q1: What are B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice?

B-hIL4/hIL4RA/hIL13/hIL13RA1/hOX40/hOX40L mice are multi-target humanized mice carrying human IL4, IL4RA, IL13, IL13RA1, OX40, and OX40L targets for Th2 inflammation and immunology drug development.

Q2: Why are IL4, IL13, IL4RA, and IL13RA1 important therapeutic targets?

IL4 and IL13 signaling through IL4RA and IL13RA1 drives type 2 inflammation, IgE production, eosinophil recruitment, asthma, and atopic dermatitis biology.

Q3: Why are OX40 and OX40L included in this model?

OX40 and OX40L are costimulatory molecules in the TNF superfamily that regulate T cell activation and inflammatory immune responses, making them important targets for allergic and autoimmune disease research.

Q4: Can this model be used for antibody efficacy studies?

Yes. The model supports in vivo evaluation of anti-human IL4RA, anti-human IL13, and anti-human OX40L antibodies in AD and OVA-induced asthma-like models.

Q5: What are the main applications of this model?

Applications include atopic dermatitis studies, asthma model studies, Th2 inflammation research, anti-human IL4RA antibody evaluation, anti-human IL13 antibody evaluation, anti-human OX40L antibody evaluation, and combination therapy development.