Biocytogen offers robust and validated asthma models induced through various methods. Notably, we have developed a series of target-humanized mice, including B-hTSLP/hTSLPR and B-hIL4/hIL4RA mice, to facilitate the evaluation of asthma therapies.
on this page
Asthma is a chronic inflammatory disorder of the airway characterized by variable airflow obstruction, airway hyperresponsiveness (AHR) and airway inflammation. The main clinical symptoms of asthma are shortness of breath, wheezing, coughing, and increased mucus secretions upon exposure to allergens. The pathogenesis of asthma is caused by complex interactions between genetic, epigenetic, and environmental factors. Pathologic changes are mediated by several types of airway cells and immune cells, including airway epithelial cells, eosinophils, and T cell subsets. In particular, Th2 cells have been thought to predominate in high eosinophilic asthma, which is characterized by increased levels of IL-4, IL-5, and IL-13.
Biocytogen provides robust and validated asthma models that can be induced by various means. In our OVA model, mice are sensitized by multiple intraperitoneal injections of ovalbumin and then challenged by inhalation of aerosolized ovalbumin. The HDM (house dust mite) and alternaria-induced asthma models involves intranasal challenges spanning a period of 4 weeks. Induction by these means results in increased IgE and eosinophil levels, while histologic staining shows increased airway mucus and inflammatory leukocyte infiltration.
| Readout | ||
| Included tests | Bronchoalveolar Lavage Fluid (BALF) | Cell numbers of Neutrophils, eosinophils, and macrophages |
| Serum | IgE level | |
| Histopathology | Bronchial mucus | |
| Immune infiltration | ||
| Histology scores | ||
| Optional tests | BALF | Total IgE, IL-4, IL-5, IL-13, TARC… |
| Lung tissue homogenate | IL-4, IL-5, IL-13, TARC… | |
| Lung tissue | IHC | |
| Airway function testing | Enhanced Pause (Penh) | |
Immune Cell Infiltration in Bronchoalveolar Lavage Fluid (BALF) of Asthmatic Mice
Increased Immune Cell number in BALF of OVA-induced mice compared with controls. BALF was collected at the end of the experiment and CD45+ cells number (A), eosinophils number (B) and the percent of eosinophils in CD45+ cells (C) were measured by flow cytometry.
IgE Induction in Serum of Asthmatic Mice
Increased IgE levels in serum of OVA-induced mice compared with controls. Serum was isolated at the end of the experiment and concentrations of OVA-specific IgE (A) and serum total IgE (B) were measured using ELISA.
Airway Histology in Asthmatic Mouse Model
H&E staining in the lungs of asthmatic mice. In contrast to the G1 untreated group, the OVA-treated G2 model animals showed asthma-related pathological changes as demonstrated by vascular and peribronchial mixed inflammatory cell infiltration (b) and mucus (a) formation in some bronchi. The above results indicate that OVA could successfully induces asthma in wild-type C57BL/6 mice.
Quantification of Immune Cells in Bronchoalveolar Lavage Fluid (BALF) Of Asthmatic Mice. Asthma was induced in wild-type BALB/c mice using OVA. (A) number of CD45+ cells in BALF; (B) number of eosinophils in BALF; (C) frequency of eosinophils in the CD45+ cell population after sensitization and challenge with OVA.
The white cell count of mice in the G2 model group was significantly increased compared with the G1 control group, and the number and proportion of eosinophils were significantly increased, suggesting that the model was successfully established. The number of CD45+ cells and eosinophils were significantly decreased in asthmatic animals treated with higher levels of dexamethasone compared with the untreated asthmatic mice (G2) group.
Airway Histology in Asthmatic Mouse Model
H&E staining in the lungs of asthmatic mice. The results showed that there was no inflammatory reaction in the pulmonary airway of G1 control group. Vascular and peribronchial inflammation was significantly increased and mucus secretion levels were increased in the G2 (OVA only) group, suggesting successful modeling of the disease. Administration of different concentrations of dexamethasone to groups G3, reduced inflammatory infiltration and mucus secretion. The results demonstrate that the OVA-induced mouse asthma model can be successfully used to verify the efficacy of corticosteroid immunosuppressive agents.
IgE Induction in Serum of Asthmatic Mice
Detection of IgE levels in serum of asthmatic mice. Serum was taken at the experimental endpoint and concentrations of OVA-specific IgE (A) and total IgE (B) were measured using ELISA.
The serum levels of OVA-specific IgE and total IgE in the G2 model group were significantly increased compared with those in the G1 control group, suggesting that the model was successfully established. After administration of different concentrations of dexamethasone (Dex), the specific IgE and total IgE levels were significantly reduced in a dose-dependent manner compared with the G2 group.
The asthma model was induced in B-hIL4/hIL4RA mice using OVA. ( A) The number of CD45 + cells in BALF. (B) The number of eosinophils in BALF. (C) The proportion of eosinophils to CD45 + cells. The results showed that after sensitization and challenge with OVA, the leukocyte infiltration of mice in G2 model group was significantly increased compared with G1 control group, and their eosinophil content was significantly increased, suggesting that the model was successfully established. After administration of dupilumab (in house), the numbers of CD45 + cells and eosinophils were significantly lower compared with the G2 model group.
Detection of IgE levels in serum of asthmatic mouse model. Serum was taken at the end of the experiment and OVA-specific IgE and total IgE levels were measured using ELISA. (A) OVA-specific IgE levels in serum. (B) Total IgE levels in serum. The results showed that the levels of OVA-specific IgE and total IgE in G2 model group were significantly increased compared with G1 control group, suggesting successful modeling. Specific and total IgE levels were significantly lower after administration of dupilumab (in house) drug compared with the G2 modeling group.
Analysis of H&E staining in the lungs of asthmatic mouse models. The results showed that there was no inflammatory cell infiltration in the pulmonary airway of G1 control group. Vascular and peribronchial inflammation (b) were significantly increased and mucus (a) secretion levels were increased in the G2 modeling group, suggesting successful modeling. G3 showed decreased inflammatory infiltration and mucus secretion in dupilumab (in house) -treated mice. The above results demonstrated that OVA could successfully establish an asthma model in B-hIL4/hIL4RA mice for efficacy evaluation.
DSI’s Buxco FinePointe Series WBP sites monitor the respiration of conscious, unrestrained animals. The animal is placed in a chamber and allowed to breathe naturally, unrestrained and untethered. The system measures the tiny air flow which is exchanged in and out of the entire chamber due to the animal’s respiration (called box flow). By applying Boyle’s Law, the box flow can be correlated to the animal’s respiratory flow. Using special algorithms, FinePointe software analyzes the box flow for many respiratory applications including tidal parameters, indications of bronchoconstriction, indications of airway irritation, and cough analysis. Since data is collected without the stress of restraint, surgery or effects of anesthesia, this technique may be the best choice for longitudinal studies, studies requiring long-term measurement, or high throughput
40-60 min for airway function testing of 2-4 mice, one test.
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.
| Readout | ||
| Included tests | Bronchoalveolar Lavage Fluid (BALF) | Cell numbers of Neutrophils, eosinophils, and macrophages |
| Serum | IgE level | |
| Histopathology | Bronchial mucus | |
| Immune infiltration | ||
| Histology scores | ||
| Optional tests | BALF | Total IgE, IL-4, IL-5, IL-13, TARC… |
| Lung tissue homogenate | IL-4, IL-5, IL-13, TARC… | |
| Lung tissue | IHC | |
| Airway function testing | Enhanced Pause (Penh) | |
Immune Cell Infiltration in Bronchoalveolar Lavage Fluid (BALF) of Asthmatic Mice
The asthma model was induced in B-hIL4/hIL4RA mice using OVA. ( A) The number of CD45 + cells in BALF. (B) The number of eosinophils in BALF. (C) The proportion of eosinophils to CD45 + cells. The results showed that after sensitization and challenge with OVA, the leukocyte infiltration of mice in G2 model group was significantly increased compared with G1 control group, and their eosinophil content was significantly increased, suggesting that the model was successfully established. After administration of dupilumab (in house), the numbers of CD45 + cells and eosinophils were significantly lower compared with the G2 model group.
IgE Induction in Serum of Asthmatic Mice
Increased IgE levels in serum of HDM-induced mice compared with controls. Serum was isolated at the end of the experiment and concentrations of total IgE were measured using ELISA.
H&E staining in the lungs of asthmatic mice. In contrast to the G1 untreated group, the HDM-treated G2 model animals showed asthma-related pathological changes as demonstrated by vascular and peribronchial mixed inflammatory cell infiltration (b) and mucus (a) formation in some bronchi. The above results indicate that HDM could successfully induces asthma in wild-type C57BL/6 mice.
Efficacy Evaluation of the anti-human IL4R (Dupilumab) in HDM-induced Asthma Model of B-hIL4/hIL4R mice. ( A) The number of CD45+ cells in BALF. (B) The number of eosinophils in BALF. (C) The proportion of eosinophils to CD45+ cells. The results showed that after sensitization and challenge with HDM, the leukocyte infiltration of mice in G4 model group was significantly increased compared with G3 control group, and their eosinophil content was significantly increased, suggesting that the model was successfully established. After administration of 25 mg/kg dupilumab (in house), the numbers of CD45+ cells and eosinophils were significantly lower compared with the G3 model group. (D) Serum was taken at the end of the experiment and total IgE levels were measured using ELISA in serum. The results showed that the levels of total IgE in G4 model group were significantly increased compared with G3 control group, suggesting successful modeling. Total IgE levels were significantly lower after administration of dupilumab (in house) drug compared with the G5 modeling group.
H&E staining in the lungs of asthmatic mice.
In contrast to the G1 untreated group, the TSLP/OVA-treated G2 model animals showed asthma-related pathological changes as demonstrated by vascular and peribronchial mixed inflammatory cell infiltration(b) and mucus (a) formation in some bronchi.
| Readout | ||
| Included tests | Bronchoalveolar Lavage Fluid (BALF) | Cell numbers of Neutrophils, eosinophils, and macrophages |
| Serum | IgE level | |
| Histopathology | Bronchial mucus | |
| Immune cell infiltration | ||
| Histology scores | ||
| Optional tests | BALF | Total IgE, IL-4, IL-5, IL-13, TARC… |
| Lung tissue homogenate | IL-4, IL-5, IL-13, TARC… | |
| Lung tissue | IHC | |
| Airway function testing | Enhanced Pause (Penh) | |
Efficacy Evaluation of the anti-human TSLP (Tezepelumab) in hTSLP/OVA-induced Asthma Model of B-TSLP/TSLPR mice. (A) The number of CD45+ cells in BALF. (B) The number of eosinophils in BALF. (C) The proportion of eosinophils to CD45+ cells. The results showed that after sensitization and challenge with OVA, the leukocyte infiltration of mice in G2 model group was significantly increased compared with G1 control group, and their eosinophil content was significantly increased, suggesting that the model was successfully established. After administration of dupilumab (in house), the numbers of CD45 + cells and eosinophils were significantly lower compared with the G2 model group. (D) BALF was isolated at the end of the experiment and concentrations of mouse TARC were measured using ELISA. (E) Serum was isolated at the end of the experiment and concentrations of mouse OVA Specific IgE were measured using ELISA.
H&E staining in the lungs of asthmatic mice. In contrast to the G1 untreated group, the TSLP/OVA-treated G2 model animals showed asthma-related pathological changes as demonstrated by vascular and peribronchial mixed inflammatory cell infiltration(b) and mucus (a) formation in some bronchi.
