Ovalbumin In Mice Research Articles

Allergen inhalation transforms the lungs into a permissive environment for local IgE responses

Abstract Immunoglobulin E (IgE) is the primary cause of allergic diseases and must be tightly regulated. Production of IgE occurs when activated B cells undergo class switch recombination (CSR) to IgE. These key events are rare, thus questions on which B cell subsets undergo IgE CSR, where IgE CSR predominantly occurs, and which molecular and cellular signals promote IgE CSR remain unresolved. We modeled allergic asthma by intranasal administration of ragweed pollen and ovalbumin (OVA) in mice, and found that allergen inhalation induced the formation of lymphoid aggregates and production of OVA-specific IgE in the lungs. Using fluorescent mice reporting IgE switching, we revealed that allergen-sensitized lungs were a major site of IgE CSR with IgG1 +memory B cells (MBCs) being the dominant IgE-switching cells. Single-cell transcriptomics and flow cytometry analyses revealed a tight correlation between CCR6 expression and lung IgG1 +MBCs, potentially explaining their recruitment/retention in the respiratory mucosa. Furthermore, lungMBCs underwent IgE CSR more frequentlythan MBCs in the draining lymph nodes or spleen. Since IgE CSR requires interleukin 4 (IL-4), we used IL-4 reporter mice and found that IL-4-producing T H2 cells were enriched in the lungs compared with lymphoid organs. Co-culture experiments using purified lung MBCs from IgE reporter mice with lung or splenic effector CD4 +T cells revealed that IgE CSR was elevated in the presence of lungT cells, suggesting lung-infiltrating T H2 cells drive elevated IgE CSR in the lungs. This study clarifies that the respiratory mucosa is a key site of IgE CSR – in allergic asthma the lung environment supports IgE CSR with IgG1 +MBCs and CD4 +T cells collaborating to promote local IgE responses. Supported by grants from NIH (F31 HL156459, T32 AI007508, R01 HL165120, R21 AI159456)

Erdosteine effects cell barriers causing decreased inflammation and increased pulmonary function in asthmatic mice exposed to nanoparticulate pollution

Objectives: Erdosteine, an oral mucoactive anti-oxidant molecule, interferes with the pathological processes seen in respiratory disorders including thickened or increased mucus production, increased oxidative stress, and chronic inflammation; however, its efficacy as an asthma therapy remains to be fully clarified. Therefore, the aim of this study was to assess the effects of erdosteine on epithelial barrier function in asthma. Methods: To investigate the effects of erdosteine on cell barrier expression in a mouse model of asthma, BALB/c mice ( n = 8 per group; total of 40 mice) were exposed to saline (Sham), ovalbumin (OVA), or OVA plus TiO2 inhalation (200 μg/m3; OVA + TiO2). The mice were then treated with erdosteine orally (OVA + TiO2 + Erdos) or intraperitoneal dexamethasone (OVA + TiO2 + Dex). Bronchoalveolar lavage and histology were performed. Enhanced pause was used as an indicator of pulmonary function, and samples were collected. The effect of erdosteine on cell barrier expression was assessed by immunoblotting and immunohistochemical analyses. Results: OVA + TiO2 + erdosteine mice exhibited decreased inflammation, and mucous gland hyperplasia, and increased pulmonary function compared with OVA + TiO2 mice. Levels of claudin (CLDN)-4 and nectin-4 protein were higher in lung tissue from OVA + TiO2 mice than Sham and OVA mice, and were reduced by erdosteine treatment. In contrast, CLDN14 and CLDN18 protein levels were lower in lung tissue from OVA + TiO2 mice than those from control mice, but were increased by treatment with erdosteine. Conclusion: Cell barriers are involved in airway inflammation in asthma patients, and erdosteine reduces airway inflammation by changing the cell barrier.

Adjuvant activity of tubeimosides by mediating the local immune microenvironment.

Rhizoma Bolbostemmatis, the dry tuber of Bolbostemma paniculatum, has being used for the treatment of acute mastitis and tumors in traditional Chinese medicine. In this study, tubeimoside (TBM) I, II, and III from this drug were investigated for the adjuvant activities, structure-activity relationships (SAR), and mechanisms of action. Three TBMs significantly boosted the antigen-specific humoral and cellular immune responses and elicited both Th1/Th2 and Tc1/Tc2 responses towards ovalbumin (OVA) in mice. TBM I also remarkably facilitated mRNA and protein expression of various chemokines and cytokines in the local muscle tissues. Flow cytometry revealed that TBM I promoted the recruitment and antigen uptake of immune cells in the injected muscles, and augmented the migration and antigen transport of immune cells to the draining lymph nodes. Gene expression microarray analysis manifested that TBM I modulated immune, chemotaxis, and inflammation-related genes. The integrated analysis of network pharmacology, transcriptomics, and molecular docking predicted that TBM I exerted adjuvant activity by interaction with SYK and LYN. Further investigation verified that SYK-STAT3 signaling axis was involved in the TBM I-induced inflammatory response in the C2C12 cells. Our results for the first time demonstrated that TBMs might be promising vaccine adjuvant candidates and exert the adjuvant activity through mediating the local immune microenvironment. SAR information contributes to developing the semisynthetic saponin derivatives with adjuvant activities.

Regulating the surface topography of CpG nanoadjuvants via coordination-driven self-assembly for enhanced tumor immunotherapy.

Immunoadjuvants play a key role in enhancing the efficacy of therapeutic tumor vaccines for treating malignant and recurrent cancers. However, due to the bottleneck in the rational design and mechanistic understanding of novel adjuvants, currently available immunoadjuvants in clinical practice are very limited. To boost adjuvant design and development, herein we propose a surface topography regulatory strategy for constructing novel adjuvants with improved adjuvant properties. One of the licensed adjuvants with a well-defined molecular mechanism of immune activation, cytosine-phosphate-guanine oligodeoxynucleotides (CpG ODNs), was used as the material framework. We constructed immunostimulatory CpG nanoparticles (CpG NPs) with different surface topographies by coordination-driven self-assembly between CpG ODNs and ferrous ions. These self-assembled CpG NPs combine the biological and physical activation abilities of innate immunity and can be used as adjuvants of tumor antigens for malignant tumor immunotherapy. The experimental results showed that these CpG NPs could rapidly enter innate immune cells and remold the tumor microenvironment (TME) to enhance anti-tumor immunotherapy via (i) inducing proinflammatory cytokine production; (ii) promoting the transformation of macrophages from immunosuppressed M2 types into immunoactivated M1 types; (iii) amplifying the antigen presentation of mature dendritic cells (DCs), and (iv) activating T cells in tumor sites. Among the prepared nanostructures, pompon-shaped nanoparticles (NPpo) showed the strongest adjuvant properties and anti-tumor immunotherapeutic effect as the adjuvant of ovalbumin in melanoma-bearing mice. Overall, this work provides an effective strategy for designing novel adjuvants for activating the immunosuppressed TME to enable better cancer immunotherapy.

Possible Anti-asthmatic Effect of Iraqi Ammi majus Seeds Extract Induced by Ovalbumin in Mice


 
 
 
 Asthma is a chronic and complex condition of the respiratory tract, that occurs in both children and adults. Currently available medications have side effects; therefore, new therapies with fewer side effects are needed. Thus, present study was designed to evaluate the potential anti-asthmatic effects of alcoholic extract of Ammi majus on ovalbumin-induced asthma in mice. Forty-eight female Albino mice were randomly assigned to one of six groups with 8 animals each. Group I received distilled water as a negative control, Group II received ovalbumin as a positive control, Groups III and IV received Ammi majus orally at 64 and 128 mg/kg/day, respectively, with sensitization, and Groups V and VI received Ammi majus orally at 64 and 128 mg/kg/day, respectively, without sensitization. Bronchoalveolar lavage fluid (BALF) was obtained 24 h after the 1st challenge to measure the number of inflammatory cell counts. In addition, lung tissue was removed for histopathological examination. Oral administration of alcoholic extract of Ammi majus at both doses (64 and 128 mg/kg) significantly inhibited ovalbumin-induced increases in total and differential cell counts of eosinophile, neutrophile, monocyte, and lymphocyte) in BALF, in addition, improve histopathologic events of asthma in lung tissue of ovalbumin-induced asthma in mice. These results demonstrated that Ammi majus alcoholic extract has a potent anti-asthmatic activity that improved ovalbumin-induced asthma.
 
 
 

Lactobacillus paragasseri BBM171 Ameliorates Allergic Airway Inflammation Induced by Ovalbumin in Mice via Modulating the Th1/Th2 Balance

Supplementation with specific probiotics has been shown to improve allergic airway symptoms. This study aimed to investigate immunomodulatory effects of a potential probiotic strain isolated from breast milk, Lactobacillus paragasseri BBM171 (BBM171), in an ovalbumin (OVA)-induced allergic mouse model. OVA-sensitized and OVA-challenged BALB/c mice were orally administered live or heat-inactivated BBM171 for 48 consecutive days. After the last allergen challenge, serum immunoglobulin (Ig) levels, inflammatory cell levels in the lungs, and cytokine levels in bronchoalveolar lavage fluid (BALF) were assessed. The results showed that oral administration of live or heat-inactivated BBM171 decreased serum levels of total IgE, OVA-specific IgE, and OVA-specific IgG1, while increasing OVA-specific IgG2a and reducing the extent of airway inflammation in OVA-induced allergic mice. In addition, both live and heat-inactivated BBM171 modulated the cytokine profile in BALF to a type 1 T helper (Th1) response. Furthermore, ex vivo experiments using OVA-induced allergic mouse splenocytes showed that both live and heat-inactivated BBM171 could regulate the Th1/Th2 balance, decrease the proinflammatory cytokine interleukin (IL)-17 level, and increase the anti-inflammatory cytokine IL-10 level. Taken together, these results suggest that oral administration of live or heat-inactivated BBM171 improved allergen-induced airway inflammation symptoms by modulating the host immune response toward Th1 dominance.

The role of calcium-sensitive receptor in ovalbumin-induced airway inflammation and hyperresponsiveness in juvenile mice with asthma.

The role of the calcium-sensitive receptor (CaSR) was assessed in a juvenile mouse model of asthma induced by ovalbumin (OVA). The experiment was divided into normal control, OVA, and OVA +2.5/5mg/kg NPS2143 (a CaSR antagonist) groups. OVA induction was performed in all groups except the normal control, followed by assessing airway hyperresponsiveness (AHR) and lung pathological changes. Serum OVA-specific IgE and IgG1 were detected with an enzyme-linked immunosorbent assay (ELISA), and inflammatory cells were counted in bronchoalveolar lavage fluid (BALF). Real-time quantitative polymerase chain reaction, ELISA, and western blotting were performed to detect gene and protein expression. NPS2143 improved the OVA-induced AHR in mice, and AHR was higher in the OVA +2.5mg/kg NPS2143 group than in the OVA +5mg/kg NPS2143 group. Furthermore, NPS2143 reduced the production of OVA-specific IgE and IgG1 in serum and the number of eosinophils and lymphocytes in BALF in OVA mice with reduced CaSR expression in lung tissues. Besides, OVA-induced mice exhibited peribronchial and perivascular inflammatory cell infiltration, which was accompanied by severe goblet cell hyperplasia/hyperplasia and airway mucus hypersecretion. Furthermore, these mice exhibited increased levels of Interleukin (IL)-5, IL-13, MCP-1, and eotaxin, which were alleviated by NPS2143. The 5mg/kg NPS2143 showed more effective than the 2.5mg/kg treatment. CaSR expression was elevated in the lung tissues of OVA-induced asthmatic juvenile mice, whereas the CaSR antagonist NPS2143 reduced AHR and attenuated the inflammatory response in OVA-induced juvenile mice, possibly exerting therapeutic effects on childhood asthma.

Lactobacillus fermentum CECT5716 Alleviates the Inflammatory Response in Asthma by Regulating TLR2/TLR4 Expression.

BackgroundAsthma is a chronic disease, which is harmful to the health of the body and the quality of life. Supplementation of Lactobacillus can affect the immune environment of the lungs through the gut-lung axis. This study aimed to explore the potential regulatory targets of Lactobacillus to relieve inflammation in asthma and determine a new approach for improving asthma.MethodsA mouse ovalbumin (OVA)-induced model was constructed. OVA mice were supplemented with Lactobacillus fermentum CECT5716 by gavage. The gut microbiota composition of normal and OVA mice was analyzed using 16S ribosomal DNA identification. BALF, serum, lung tissues, and duodenal tissues were collected. Wright’s staining was performed to determine the cell content of the alveolar lavage fluid. Hematoxylin-eosin staining, Masson staining, and periodic acid-Schiff staining were performed to observe the improvement in the lungs of OVA mice supplemented with Lactobacillus. Immunofluorescence was performed to measure the severity of the intestinal barrier leakage. Enzyme-linked immunosorbent assay was carried out to determine the expression levels of inflammatory cell factors, while quantitative reverse transcription-polymerase chain reaction and western blotting were performed to detect the levels of toll-like receptor 2 (TLR2)/TLR4 expression and cell adhesion factors.ResultsCompared with Control mice, OVA mice exhibited malignant conditions, such as intestinal leakage and lung edema. After supplementation with Lactobacillus, the inflammatory cell content in the bronchoalveolar lavage fluid decreased, and the inflammatory response was alleviated. The level of TLR2/TLR4 expression was reduced. The inflammatory cell infiltration in the airway mucosa of OVA mice was improved, alveolar swelling was reduced and the basement membrane appeared thinner.ConclusionThe Lactobacillus inhibited the TLR2/TLR4 expression in OVA mice. Supplementation with Lactobacillus can alleviate the inflammatory response in OVA mice, inhibit pulmonary fibrosis, and treat asthma.

Dietary olive oil enhances the oral tolerance of the food allergen ovalbumin in mice by regulating intestinal microecological homeostasis.

Increasing evidence indicates that intestinal microecological imbalances are strongly associated with food allergen intolerance. This study investigated the effect of olive oil on food allergy susceptibility and intestinal microecology based on an ovalbumin (OVA)-sensitized mouse model. The results indicated that the allergic symptoms of sensitized mice were alleviated when they were supplemented with olive oil at 1-3g/kg per day for 7 weeks. Intestinal epithelium observation showed repaired ileum villi and upregulated tight junction (TJ) protein expression. Furthermore, the levels of the cytokines (e.g., IL-10) secreted by regulatory T cells were increased, whereas T helper 2 (Th2) cell-associated factors were decreased in lamina propria. Furthermore, 16S rRNA sequencing indicated reduced Burkholderiaceae and increased Clostridiaceae in the intestinal microflora. The results suggest that an olive oil-enriched diet may effectively prevent food allergies by regulating the intestinal microecological balance. PRACTICAL APPLICATIONS: Recent studies emphasized that intestinal ecological imbalance, including intestinal immunity and microflora structure, plays an important role in affecting the occurrence and development of food allergy. The present results implied that olive oil, one of the main components of the Mediterranean diet, can effectively ameliorate the symptoms of OVA-induced food allergy by regulating intestinal microecological homeostasis. Therefore, dietary supplementation with olive oil may be an effective strategy for preventing food allergies.

A serological biomarker of type I collagen degradation is related to a more severe, high neutrophilic, obese asthma subtype

BackgroundAsthma is a heterogeneous disease; therefore, biomarkers that can assist in the identification of subtypes and direct therapy are highly desirable. Asthma is a chronic inflammatory disease that leads to changes in the extracellular matrix (ECM) by matrix metalloproteinases (MMPs) degradation causing fragments of type I collagen that is released into circulation.ObjectiveHere, we asked if MMP-generated type I collagen (C1M) was associated with subtypes of asthma.MethodsC1M was serologically assessed at baseline in the adult participants of the All Age Asthma study (ALLIANCE) (n = 233), and in The Prospective Epidemiological Risk Factor study (PERF) (n = 283). In addition, C1M was assessed in mice sensitized to ovalbumin (OVA) and challenged with OVA aerosol. C1M was evaluated in mice with and without acute neutrophilic inflammation provoked by poly(cytidylic-inosinic) acid and mice treated with CP17, a peptide inhibiting neutrophil accumulation.ResultsSerum C1M was significantly increased in asthmatics compared to healthy controls (p = 0.0005). We found the increased C1M levels in asthmatics were related to blood neutrophil and body mass index (BMI) in the ALLIANCE cohort, which was validated in the PERF cohort. When patients were stratified into obese (BMI > 30) asthmatics with high neutrophil levels and uncontrolled asthma, this group had a significant increase in C1M compared to normal-weight (BMI < 25) asthmatics with low neutrophil levels and controlled asthma (p = 0.0277). C1M was significantly elevated in OVA mice with acute neutrophilic inflammation compared to controls (P = 0.0002) and decreased in mice treated with an inhibitor of neutrophil infiltration (p = 0.047).Conclusion & clinical relevanceC1M holds the potential to identify a subtype of asthma that relates to severity, obesity, and high neutrophils. These data suggest that C1M is linked to a subtype of overall inflammation, not only derived from the lung. The link between C1M and neutrophils were further validated in in vivo model.Trial registration(ALLIANCE, NCT02419274).

Co-Application with Tannic Acid Prevents Transdermal Sensitization to Ovalbumin in Mice.

Transdermal sensitization to allergens is of great concern as a sensitization route for food allergies. This skin-mediated invasion and sensitization to allergens is involved in skin barrier breakdown and inflammation, followed by the production of several kinds of cytokines. Cytokines such as thymic stromal lymphopoietin and thymus and activation-regulated chemokine are also involved. In this study, we investigated the suppressive effect of tannic acid (TA) on transdermal sensitization using ovalbumin (OVA), a major egg-white allergen. We also analyzed the mechanisms associated with the inhibitory effects of TA. The results showed that the co-application with TA prevents transdermal sensitization to OVA. As possible mechanisms, its anti-inflammatory and astringent effect on the skin and binding ability with the protein were considered. These results indicate that TA could be applied to cosmetics and lotions, which could suppress the transdermal sensitization to allergens.

A Phenotypic Analysis of Involucrin–Membrane-Bound Ovalbumin Mice after Adoptive Transfer of Ovalbumin-Specific CD8+ T Cells

To investigate the mechanism of autoimmunity and peripheral tolerance in the skin, several transgenic mouse strains expressing membrane-bound ovalbumin (mOVA) as an epidermal self-antigen under the control of keratinocyte-specific promotors, such as keratin 5 and keratin 14, were employed in combination with adoptive transfer of CD8+ T cells from OT-I mice (OT-I T cells) that recognize an ovalbumin-derived peptide. However, these strains showed bodyweight loss and required additional inflammatory stimuli, such as γ-irradiation and tape-stripping, to induce skin inflammation. In this study, we generated a mouse strain expressing mOVA under the control of human involucrin promoter (involucrin-mOVA mice). In contrast to previous strains, involucrin-mOVA mice spontaneously developed skin inflammation after the transfer of OT-I T cells in the absence of external stimuli without significant bodyweight loss. We focused on the skin infiltration process of OT-I T cells and found that transferred OT-I T cells accumulated around the hair follicles in the early phase of skin inflammation, and in the later phase, the skin inflammation spontaneously resolved despite the remaining OT-I T cells in the skin. Our involucrin-mOVA mice will provide a promising tool to investigate the pathogenesis and the tolerance mechanisms of cytotoxic skin autoimmunity.

Effect of Echinococcus granulosus hydatid cyst fluid protein on allergic rhinitis induced by ovalbumin in mice

To investigate the protective effect of Echinococcus granulosus hydatid cyst fluid protein (HCFP) on ovalbumin (OVA)-induced allergic rhinitis (AR) in mice. Twenty-four BALB/c mice at ages of 8 to 10 weeks, each weighing approximately 20 g, were randomly divided into four groups, including groups A (blank control group), B (blank intervention group), C (AR model group) and D (AR+HCFP intervention group), with 6 mice in each group. On days 0, 2, 4, 6, 8, 10 and 12, mice in groups A, B, C and D were injected with 200 μL sterile phosphate buffered saline (PBS), 200 μL sterile PBS containing 20 μg HCFP, 200 μL sterile PBS containing 50 μg OVA and 5 mg Al(OH)3 gel, and 200 μL sterile PBS containing 50 μg OVA, 5 mg Al(OH)3 gel and 20 μg HCFP, respectively. On days 14 to 20, mice in groups A, B, C and D were administered with 40 μL sterile PBS, 40 μL sterile PBS containing 20 μg HCFP, 40 μL sterile PBS containing 2 mg OVA and 40 μL sterile PBS containing 2 mg OVA and 20 μL HCFP by nasal drop, respectively. Mouse behavioral changes were observed and behavioral scores were estimated. The serum levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), IL-5, IL-10, transforming growth factor-β (TGF-β) and OVA-specific IgE antibody (OVA-sIgE) were measured using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of mouse nasal mucosa were observed by hematoxylin and eosin (HE) staining. The mean behavioral score was significantly greater in Group C (6.83 ± 0.50) than in groups A (1.17 ± 0.52) and B (1.33 ± 0.52) (P < 0.05), while a lower mean behavioral score was estimated in Group D (3.50 ± 0.50) than in Group C (P < 0.05). There were significant differences among the groups in terms of serum IFN-γ (F = 4.08, P < 0.05), IL-4 (F = 275.90, P < 0.05), IL-5 (F = 96.82, P < 0.05), IL-10 (F = 77.67, P < 0.05), TGF-β (F = 9.98, P < 0.05) and OVA-sIgE levels (F = 44.69, P < 0.05). The serum IFN-γ level was significantly lower in Group C than in groups A, B and C (P < 0.05), and the serum levels of IL-4, IL-5 and OVA-sIgE were significantly higher in Group C than in groups A, B and C (P < 0.05), while the serum IL-10 and TGF-β levels were significantly greater in Group D than in Group C (P < 0.05). Microscopy showed apparent loss of nasal mucosa cilia, increased number and enlargement of goblet cells, interstitial edema and submucous vascular dilation in Group C, while the pathological changes of nasal mucosa were alleviated in Group D relative to Group C. E. granulosus HCFP has a protective activity against OVA-induced allergic rhinitis in mice.

Adjuvant effect of two polysaccharides from the petals of Crocus sativus and its mechanisms

Two polysaccharides from Crocus sativus petals (PCSPs), PCSPA and PCSPB, presented immunopotentiatory activity through activating macrophages via MAPK and NF-κB pathway. In this study, two PCSPs were investigated for the adjuvant effects and underlying mechanisms using ovalbumin (OVA) in mice. PCSPA and PCSPB remarkably not only boosted the OVA-specific IgG antibody and its isotype titers, but strengthened splenocyte proliferation and natural killer cell activities in immunized mice. Both PCSPs also dramatically triggered the production of Th1- and Th2-cytokines and facilitated the gene expression of Th1- and Th2-cytokines and transcription factors in OVA-stimulated splenocytes. In mechanisms, two PCSPs rapidly elicited the gene and protein expression of cytokines and chemokines, promoted the recruitment and antigen uptake of immune cells in the injected-muscles, and augmented the migration and antigen transport of immune cells to the draining lymph nodes. These findings demonstrated that PCSPs enhanced and improved immune responses and simultaneously elicited Th1- and Th2-immune responses to OVA through activating innate immune microenvironment, and that they could act as promising vaccine adjuvant candidates.

Has2 Regulates the Development of Ovalbumin-Induced Airway Remodeling and Steroid Insensitivity in Mice.

HAS2 is a member of the gene family encoding the hyaluronan synthase 2, which can generate high-molecular-weight hyaluronan (HMW-HA). Our previous study identified HAS2 as a candidate gene for increased susceptibility to adult asthma. However, whether HAS2 dysfunction affects airway remodeling and steroid insensitivity is still limited. Therefore, this study aimed to clarify the Has2 dysfunction, triggering severe airway remodeling and steroid insensitivity in a murine model of asthma. Has2 heterozygous-deficient (Has2 +/−) mice and their wild-type littermates have been evaluated in a model of chronic ovalbumin (OVA) sensitization and challenge. Mice present a higher sensitivity to OVA and higher IL-17 release as well as eosinophilic infiltration. RNA sequencing demonstrated the downregulation of EIF2 signaling pathways, TGF-β signaling pathways, and heat shock proteins with Th17 bias in Has2 +/−-OVA mice. The combined treatment with anti-IL-17A antibody and dexamethasone reduces steroid insensitivity in Has2 +/−-OVA mice. Has2 attenuation worsens eosinophilic airway inflammation, airway remodeling, and steroid insensitivity. These data highlight that HAS2 and HMW-HA are important for controlling intractable eosinophilic airway inflammation and remodeling and could potentially be exploited for their therapeutic benefits in patients with asthma.

Development and Evaluation of an Acute Asthma Model Induced by Ovalbumin and Formaldehyde Exposure in Murine

Asthma is a common prolonged inflammatory disease of the upper respiratory tract, affecting millions of people in all age groups. At present, the treatment of asthma has become a challenge for practitioners because of the regular increment in air pollution worldwide. Subsequently, during the new drug development process, preclinical assessment of active compounds through a quick and robust model is urgently required. Thus, this study aimed to establish an acute asthma model using ovalbumin and formaldehyde in mice. In this experimental study, all animals were sensitized with ovalbumin on 0th and 5th d and formaldehyde on the 3rd, 4th, 6th, and 7th d followed by an ovalbumin challenge from 8th to 10th d. During induction of disease, individual groups received beclomethasone and rutin as standard and test drugs, respectively. At the end of the study, the whole lung was harvested, weighed, and assessed for lung weight index. Also, oxidative stress and inflammatory biomarkers were estimated, and lung histopathology was performed. In the results, it was observed that the lung weight index, total leukocyte count and differential leukocyte count, and malondialdehyde values, significantly (p<0.05) decreased, and superoxide dismutase, glutathione and catalase activity were markedly (p<0.05) increased in the rutin treated group as compared to the diseased group. Histopathological studies of lung tissue in the diseased group showed extensive perivascular edema. However, in the rutin-treated group, mucus gland hyperplasia was significantly decreased. Therefore, in this experimental research, an attempt was made to develop a successful acute asthma model within a short period, which reversed on repeated dosing with rutin in murine.

Caspase-1-Dependent Pyroptosis Mediates Adjuvant Activity of Platycodin D as an Adjuvant for Intramuscular Vaccines

Platycodin D (PD) is a potent adjuvant with dual Th1 and Th2 potentiating activity, but its mechanisms of action remain unclear. Here, the C2C12 myoblast cell line and mice were used as in vitro and in vivo models to identify potential signaling pathways involved in the adjuvant activity of PD. PD induced a transient cytotoxicity and inflammatory response in the C2C12 cells and in mouse quadricep muscles. A comparative analysis of microarray data revealed that PD induced similar gene expression profiles in the C2C12 cells and in the quadricep muscles, and triggered rapid regulation of death, immune, and inflammation-related genes, both in vivo and in vitro. It was further demonstrated that caspase-1-dependent pyroptosis was involved in the PD-induced cytotoxicity and inflammatory response in the C2C12 cells via the Ca2+–c-jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (MAPK)–NLR family pyrin domain containing 3 (NLRP3) inflammasome signaling pathway. Consistently, the in vivo analysis revealed that a local blockage of NLRP3 and caspase-1 inhibited PD-induced cytokine production and immune cell recruitment at the injection site, and impaired the adjuvant activity of PD on antigen-specific immune responses to model antigen ovalbumin (OVA) in mice. These findings identified the caspase-1-dependent adjuvanticity of PD and expanded the current knowledge on the mechanisms of action of saponin-based adjuvants.

The mechanism of rapamycin in promoting asthmatic regulatory T cell differentiation and function.

To investigate the mechanism of rapamycin in promoting asthmatic regulatory T cell differentiation . Asthma model was prepared by sensitization and challenge of ovalbumin in mice. Spleen CD4CD25 T cells were sorted from the asthmatic mice and normal mice by ultrahigh speed flow cytometer, and divided into three groups. Transforming growth factor-β and interleukin-2, or combined with rapamycin (final concentration of 500 nmol/L) were given in the model group or the rapamycin group. The levels of Treg cells and CD4CD25 T cells were detected by flow cytometry. The phosphorylation level of downstream proteins of S6 and Akt in the mTORC1/2 signaling pathway were examined by Western blotting. Compared with the model group, the differentiation level of Treg cells in the rapamycin group was significantly increased, the proliferation level of CD4CD25 T cells was decreased, and the phosphorylations of the mTORC1/2 substrates, S6 protein and Akt were decreased （all <0.05）. Rapamycin can promote the differentiation and function of Treg cells via inhibition of the mTORC1/2 signaling pathway.

Effect of parasympathetic inhibition on expression of ILC2 cells in a mouse model of allergic rhinitis

BackgroundWe wanted to investigate whether parasympathetic inhibition affected the expression of type 2 innate lymphoid cells (ILC2s) in the nasal mucosa of a mouse model of allergic rhinitis (AR). MethodsThirty male C57BL/6 mice were randomly divided into 3 groups: control group, AR group, AR-treated group. AR nasal symptoms were assessed on a semi-quantitative scale according to the frequencies of nose rubbing and sneezing and the degree of rhinorrhea. The expression of cytokines protein in serum was detected by enzyme linked immunosorbent assay (ELISA). The number of ILC2s in nasal mucosa was detected by immunofluorescence double staining assay. Quantitative real-time Polymerase Chain Reaction (qPCR) was used to detect the expression of ILC2-associated factor in nasal mucosa. ResultsThe symptom scores of the AR group were significantly higher than those of the control group and AR-treated group. The expression levels of mouse ovalbumin (OVA) specific IgE, IL4, IL5, and IL13 in the serum of AR group were significantly higher than those in the control group and AR-treated group. The number of ILC2s and the gene expression of ILC2s related factors GATA3, CD25 and CD90 (Thy1) in the nasal mucosa of the AR group were significantly higher than those of the control group and AR-treated group. ConclusionsWe found that parasympathetic inhibition relieved AR symptoms and inhibited immune response of AR mice. ILC2s play an important role in the occurrence and development of AR, and parasympathetic nerve inhibition reduced the number of ILC2s and inhibited the cytokines expression by ILC2s. Our data provide information on the mechanism of action of parasympathetic inhibition in AR.

Effect of Vitamin D Deficiency and Supplementation in Lactation and Early Life on Allergic Airway Inflammation and the Expression of Autophagy-Related Genes in an Ovalbumin Mouse Model.

Background and ObjectiveVitamin D is involved in various physiological and pathological processes, including inflammation and autophagy. We aimed to investigate the effects of dietary vitamin D deficiency or supplementation initiated in lactation and early life on inflammation and autophagy in an ovalbumin (OVA) mouse model.MethodsFemale BALB/c were fed with vitamin D-deficient, sufficient or supplemented diets throughout lactation and their offspring followed the same diet after weaning. Offspring were then sensitized and challenged with OVA, airway resistance (RL) was measured, and their serum, bronchoalveolar lavage fluid (BALF), and lung tissue were collected. Alveolar macrophages (AMs) were isolated from lung tissue and cultured with different concentrations of 1,25(OH)2D3. The expressions of autophagy-related (ATG) proteins including light-chain 3 (LC3), Beclin-1, and ATG5, and NF-κB p65 in lung tissue and AMs were measured.ResultsOVA sensitization and challenge induced dramatic allergic airway inflammation and higher RL in the vitamin D-deficient group compared with vitamin D-sufficient or the supplemented group. The expression of ATGs including LC3, Beclin-1, and ATG5, and NF-κB p65 in lung tissue in the vitamin D-deficient OVA-mediated group was increased compared with vitamin D-supplemented OVA-mediated group. There was correlation between the expression of LC3 mRNA and inflammatory cell numbers and cytokines in BALF. In vitro, 1,25(OH)2D3 also regulated the expression of LC3, Beclin-1, ATG5, and NF-κB p65 mRNA in AMs in a time- and dose-dependent manner.ConclusionDeficiency of vitamin D in early life may aggravate allergic airway inflammation, and maintaining sufficient vitamin D during early life is necessary for lung health. Vitamin D may modulate autophagy in lungs of OVA sensitized/challenged mice, thus playing a protective role in OVA-induced allergic airway inflammation.