OBJECTIVETo observe the early and late symptomatic, pathological and immunological changes in an intranasal ovalbumin-induced animal model of allergic rhinitis in guinea pigs.

METHODSGuinea pigs were intranasally sensitized with ovalbumin absorbed on aluminum hydroxide and after 5 days' interval, they were challenged with 1% ovalbumin solution once every 3 days for total 11 times. Two control groups were studied in parallel, the positive treatment control group was treated with antihistamine and the negative control group was sham-sensitized and sham-challenged. Typical symptoms of allergic rhinitis, such as sneezing, nasal scratching, nasal blockage and rhinorrhea were evaluated. Passive cutaneous anaphylaxis reaction (PCA) was performed to measure the levels of IgG1 and IgE. Eosinophils infiltration and goblet cells in nasal mucosa were observed. In addition, the level of histamine and the number of total leukocytes and eosinophils in the nasal lavage fluid were also measured.

RESULTSIn the model group, symptoms of sneezing, nasal scratching, nasal blockage and rhinorrhea were induced after ovalbumin challenge. The respiratory rate (RR), which reflected the resistance of upper airway, showed a biphasic change. In the PCA test, IgG1 and IgE levels increased after challenges. Eosinophil infiltration in nasal mucosa was more obvious in active groups in comparison to with the negative control group (P < 0.05 or < 0.01). The histamine, total leucocytes and eosinophils levels in nasal lavage fluid also showed higher in the model group (P < 0.05 or < 0.01). The antihistamine treated animals were also induced out above changes but modest compared with the model group (P < 0.05 or < 0.01). The negative control showed few of above changes with significant difference (P < 0.05 or < 0.01).

CONCLUSIONSOur results implied that the modified animal model of allergic rhinitis was capable of showing satisfactory symptomatic and pathophysiological changes in allergic rhinitis. It showed a biphasic nasal blockage with shorter establishment duration. The model also had good treatment reaction to antihistamine. The animal model we introduced may be useful in the study of allergic rhinitis.

Administration, Intranasal ; Animals ; Disease Models, Animal ; Guinea Pigs ; Nasal Lavage Fluid ; Ovalbumin ; administration & dosage ; Rhinitis, Allergic, Perennial

We have investigated whether oral administration of ovalbumin (OVA) could prevent active systemic anaphylaxis to the antigen. Oral tolerance was induced by a single feecfing with 40 mg OVA before, but not after, sensitization characterized by diminished OVA-specific IgE and IgG responses. Feeding 15 mg OVA suppressed anaphylaxis and antibody responses to a lesser extent. Spleen cells from tolerant donors were incapable of transferring the tolerance to naive recipients. Pretreatment of cyclophosphamide (100 mg/kg) 2 days before OVA feeding did not restore the tolerance. Furthermore, in vitro cell mixing studies showed that the proliferation of spleen cells from OVA- sensitized donors was not inhibited by the addition of spleen cells from tolerant donors, arguing against the role of suppressor cells. Anergy was demonstrated by the ability to reverse the tolerant state after culturing tolerant cells in the presence of IL-2. These findings indicate that only a high-dose (40 mg) feeding OVA was found to be effective in inducing tolerance in this experimental system, and demonstrate anergy as the mechanism underlying oral tolerance to systemic anaphylaxis.
Administration, Oral ; Anaphylaxis* ; Antibody Formation ; Cyclophosphamide ; Humans ; Immunoglobulin E ; Immunoglobulin G ; Interleukin-2 ; Ovalbumin ; Ovum ; Spleen ; Tissue Donors

OBJECTIVES: The aim of this study was to investigate whether the ciliary activity of respiratory epithelium is affected in allergic rhinitis. METHODS: Twenty Wistar rats were divided into an unsensitized control group and sensitized allergic group. The sensitized group was immunized intraperitoneally with ovalbumin, followed by intranasal administration of ovalbumin. Allergy was determined by an increase in nasal symptoms, the number of tissue eosinophils and a positive result to a passive cutaneous anaphylaxis (PCA) test. Nasal, nasopharyneal, tracheal, and bronchial epithelial cells were obtained from both the control and allergic groups. Ciliary beat frequency (CBF) was measured using a video-computerized analysis technique in vitro. We compared the CBF of two groups in each site. We also evaluated the findings of the nasal mucosa of both groups with an scanning electron microscope. RESULTS: In vitro CBF measurement demonstrated that the CBF of the control and allergic groups did not differ significantly (p>0.05). CONCLUSION: CBF is not affected by respiratory allergy.
Administration, Intranasal ; Animals ; Eosinophils ; Epithelial Cells* ; Hypersensitivity ; Nasal Mucosa ; Ovalbumin ; Passive Cutaneous Anaphylaxis ; Rats* ; Rats, Wistar ; Respiratory Mucosa ; Rhinitis*

PURPOSE: We used the sensitized rat model to study the effect of the inflammation evoked by immune response on the smooth muscles of the urinary bladder, trachea and intestine and changes of mast cell. MATERIALS AND METHODS: The rats were divided into 3 groups (group 1: control - not-sensitized. no antigen challenge, group 2: sensitized, no antigen challenge, group 3: sensitized, antigen challenge). Each group was divided into two subgroups of male and female. Rats were sensitized by intraperitoneal injection of ovalbumin (10mg/ml/kg) given on days 1, 3 and 5, 4 weeks after last injection, sensitized rats were treated with intravesical ovalbumin for 1 hour (antigen challenge) and controls runned simultaneously with the unsensitized rats by intravesical instillation of normal saline. After intravesical instillation contractility and number of mast cells of each organ were measured. RESULTS: Only female-sensitized group showed significantly increased contractility of bladder muscle (p<0.05) while no significant difference in trachea and intestine among groups. In male rats there were no significant differences between groups and organs. In male and female groups, there was increased contractility in female rats than male but showed no significant difference between sexes. In histologic study, sensitized, ovalbumin group showed more mast cells in detrusor muscle and submucosa than non-sensitized, saline group in both male and female rats. Also there were significant differences in female rats than male rats. Detrusor to submucosa mast cell ratio was significantly higher in sensitized, ovalbumin group. CONCLUSIONS: We concluded that there might be some other factors that made differences in response to antigen challenge between sexes and among the organs. We observed increased contractility in female rat bladder and this may explain higher incidence of interstitial cystitis in female than male.
Administration, Intravesical ; Animals ; Cystitis, Interstitial ; Female ; Humans ; Incidence ; Inflammation ; Injections, Intraperitoneal ; Intestines* ; Male ; Mast Cells* ; Models, Animal ; Muscle, Smooth* ; Ovalbumin ; Rats* ; Trachea* ; Urinary Bladder*

EPSAH is an exopolysaccharide from Aphanothece halophytica GR02. The present study was designed to evaluate its toxicity and adjuvant potential in the specific cellular and humoral immune responses in ovalbumin (OVA) in mice. EPSAH did not cause any mortality and side effects when the mice were administered subcutaneously twice at the dose of 50 mg·kg(-1). Hemolytic activity in vitro indicated that EPSAH was non-hemolytic. Splenocyte proliferation in vitro was assayed with different concentrations of EPSAH. The mice were immunized subcutaneously with OVA 0.1 mg alone or with OVA 0.1 mg dissolved in saline containing Alum (0.2 mg) or EPSAH (0.2, 0.4, or 0.8 mg) on Day 1 and 15. Two weeks later, splenocyte proliferation, natural killer (NK) cell activity, production of cytokines IL-2 from splenocytes, and serum OVA-specific antibody titers were measured. Phagocytic activity, production of pro-inflammatory cytokines IL-1 and IL-12 in mice peritoneal macrophages were also determined. EPSAH showed a dose-dependent stimulating effect on mitogen-induced proliferation. The Con A-, LPS-, and OVA-induced splenocyte proliferation and the serum OVA-specific IgG, IgG1, and IgG2a antibody titers in the immunized mice were significantly enhanced. EPSAH also significantly promoted the production of Th1 cytokine IL-2. Besides, EPSAH remarkably increased the killing activities of NK cells from splenocytes in the immunized mice. In addition, EPSAH enhanced phagocytic activity and the generation of pro-inflammatory cytokines IL-1 and IL-12 in macrophages. These results indicated that EPSAH had a strong potential to increase both cellular and humoral immune responses, particularly promoting the development of Th1 polarization.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Cyanobacteria ; chemistry ; Female ; Immunity, Cellular ; Immunity, Humoral ; Immunization ; Interleukin-12 ; immunology ; Interleukin-2 ; immunology ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred ICR ; Ovalbumin ; immunology ; Polysaccharides ; administration & dosage ; immunology ; Rabbits ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo study the effect of neonatal immunization with different dosage allergen on the immunity of mice when grown.

METHODSFifty neonatal BALB/c mice were divided into 4 groups randomly and subcutaneous injected with different dosage of ovalbumin (OVA) on day 1, 8 and 15 after born [NS group(10): injected with saline alone; NS + AL group (10): injected with saline and AL(OH)3; small dosage (SD) group (15): injected with 10 microg OVA and AL(OH)3; large dosage (LD) group (15): injected with 1000 microg OVA and AL(OH)3]. The mice were then challenged using caudal vein injection on 5 weeks old (NS group and NS + AL group were challenged with saline, SD group and LD group were challenged with 100 microg OVA). The blood was collected 1 week later to examine OVA specific IgE, IgG1 and IgG2a. Mononuclear cells were drawn from the spleen and cultured. Concentration of IL-4, IFN-r, IL-10 was examined in the cultural supernatant. Flow cytometry was used to test the expression of CD4+ IL-4+, CD4+ IFN-gamma+, CD4 IL-10 cells.

RESULTSIt was found that concentration of OVA specific IgE (OVA-sIgE) in SD group (0.33 +/- 0.18) was significantly higher than that of NS (0.07 +/- 0.01) and NS + AL (0.09 +/- 0.04) group (t value was -3.46 and -3.21, all P < 0.01), and LD group (0.17 +/- 0.10) as well (t = 2.58, P < 0.05). The concentration of OVA-sIgE was higher in LD group than that of NS group (t = -2.53, P < 0.05), but similar with that of NS + AL group (t = -2.04, P > 0.05). Both the concentration of OVA-sIgG1 and sIgG2a was higher in SD and LD group than that of NS and NS + AL group (all P < 0.05). The concentration of IL-4, IFN-gamma and IL-10 in the cultural supernatant of spleen mononuclear was all higher in SD group than that of NS and NS + AL group (all P < 0.01). The ratio of IFN-gamma/IL-4 was significantly lower in SD group than that of NS and NS + AL group (t value was 2.14, 3.44, all P < 0.05), while the same ratio was higher in LD group than that of NS and NS + AL group (t value was -2.14, -1.61, all P < 0.05). Ratio of CD4+ IL-4+ cells was significant lower in LD group than that of SD group (P < 0.05), while it was not different with that of NS and NS + AL group (P > 0. 05).

CONCLUSIONNeonatal immunization with low dosage OVA could generate a specific immunity with Th2 direction, while with large dosage OVA could generate a specific immunity with Th1 direction.

Allergens ; administration & dosage ; Animals ; CD4-Positive T-Lymphocytes ; Dose-Response Relationship, Drug ; Immunization ; Immunoglobulin G ; metabolism ; Interferon-gamma ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; administration & dosage

AIMTo investigate the effect of inhalation of cyclosporin (CsA) on antigen-induced airway inflammation in Sprague-Dawley rats.

METHODSRats were sensitized with antigen (ovalbumin, OA). After two weeks, the sensitized rats were pretreated with aerosol CsA (5, 10, 20 g x L(-1)), once per day for 7 days. Then, the sensitized rats were challenged with OA (10 g x L(-1), once per day) for 2 days at day 20 after sensitization. The number of eosinophils in bronchoalveolar lavage fluid (BALF) and peripheral blood, histological changes of lung tissue, and TNF-alpha content in BALF were investigated.

RESULTSInhalation of CsA significantly reduced the number of eosinophils in BALF and peripheral blood, inflammatory infiltration and tissue edema of lung tissue, decreased the content of TNF-alpha in BALF.

CONCLUSIONInhalation of CsA inhibited airway inflammation in rats, and the mechanism is related to inhibition of TNF-alpha release.

Administration, Inhalation ; Animals ; Asthma ; chemically induced ; metabolism ; pathology ; Bronchoalveolar Lavage Fluid ; chemistry ; Cyclosporine ; administration & dosage ; pharmacology ; Eosinophils ; pathology ; Female ; Immunosuppressive Agents ; pharmacology ; Leukocyte Count ; Lung ; pathology ; Male ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVEInhaled glucocorticosteroids (ICS) remains the first line controller medication for chronic airway inflammation in asthma till now. If the impact of allergen could not be eliminated, how would the improvement of airway inflammation be achieved with inhaled glucocorticosteroids therapy? What was its effect on airway remodeling? In this study, an animal model of asthma was established and the effects of budesonide on airway allergic inflammation and extracellular matrix (ECM) deposition in sensitized guinea pigs with repeated exposure to allergen were investigated.

METHODSThirty-two male Hartley guinea pigs were randomly divided into four groups with 8 in each group: (A) Group of repeated exposure to ovalbumin (OVA), (B) Group of repeated exposure to OVA plus budesonide (BUD) intervention, (C) Group of stopping repeated exposure to OVA plus stopping BUD intervention, (D) Control group. At 24 h after the last OVA challenge (8 weeks after the first OVA challenge), bronchoalveolar lavage fluid (BALF) was collected from each animal. Total and differential leukocyte counts in BALF was performed on cell suspension smear stained with May-Grünwald-Giemsa (MGG) method. The upper lobe of right lung was removed and regularly fixed, then paraffin embedded lung tissues sections were prepared. The count of eosinophils infiltrated in the airway wall was performed on H&E stained lung tissue sections with LEICA Q500IW computerized image analysis system. Fibronectin and collagen type III (Col-III) deposited in the airway wall were detected by immunohistochemical staining on the paraffin embedded lung tissues sections. The intensity of positive reaction of fibronectin or Col-III deposited in the airway wall was analyzed with LEICA Q500IW computerized image analysis system.

RESULTSThe count of eosinophils in BALF (x 10(5)/ml) of group A and B were higher than that of group C and D (35.70 +/- 25.22, 11.49 +/- 5.51 vs. 1.00 +/- 0.90, 1.02 +/- 0.78, P < 0.01), the difference between group A and B, group B and C was significant. The count of eosinophils infiltrated at each level of airway wall in group A and B were higher than that of group C and D (large airway: 6.95 +/- 2.28, 1.54 +/- 1.09 vs. 0.76 +/- 0.45, 0.88 +/- 0.25; medial airway: 9.22 +/- 3.89, 3.99 +/- 2.3 vs. 1.25 +/- 1.20, 0.64 +/- 0.36; small airway: 11.56 +/- 4.02, 2.67 +/- 1.15 vs. 1.32 +/- 0.83, 0.43 +/- 0.24, P < 0.01), the difference between group A and B, group B and C was significant. The gray values of fibronectin deposited in medial and small airway of group A and B were lower than those of group C and D (medial airway 122 +/- 22, 174 +/- 23 vs. 219 +/- 34, 229 +/- 20; small airway 135 +/- 29, 165 +/- 41 vs. 236 +/- 20, 220 +/- 16, P < 0.05), the difference between group A and B, group B and C was significant. The gray values of Col-III deposited in medial and small airway of group A and B were lower than those of group C and D (medial airway 153 +/- 21, 174 +/- 22 vs. 189 +/- 14, 200 +/- 18; small airway 133 +/- 23, 176 +/- 20 vs. 191 +/- 14, 198 +/- 20, P < 0.05), the difference between group A and B was significant.

CONCLUSIONInhaled budesonide could partially inhibit allergic inflammation and ECM deposition in airway wall in guinea pig chronic asthma model with repeated exposure to allergen. Early inhaled budesonide combined with avoidance of OVA exposure could completely inhibit allergic inflammation and ECM deposition. These results suggest that the inhibitory effect on airway allergic inflammation and airway remodeling of inhaled glucocorticosteroids would be limited when the allergen factor could not be avoided.

Administration, Inhalation ; Airway Remodeling ; drug effects ; immunology ; Allergens ; administration & dosage ; immunology ; Animals ; Asthma ; chemically induced ; drug therapy ; immunology ; Bronchitis, Chronic ; chemically induced ; drug therapy ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Budesonide ; administration & dosage ; pharmacology ; Collagen Type III ; metabolism ; Disease Models, Animal ; Eosinophils ; immunology ; Extracellular Matrix ; immunology ; Fibronectins ; metabolism ; Glucocorticoids ; administration & dosage ; pharmacology ; Guinea Pigs ; Immunohistochemistry ; Lung ; drug effects ; immunology ; Male ; Ovalbumin ; administration & dosage ; immunology

PURPOSE: Asthma is a pulmonary chronic inflammatory disease characterized by airway obstruction and hyperresponsiveness. Pattern recognition receptors are known to play a key role in the development of allergic diseases as well as host defenses against microbial infection. Receptor interacting protein 2 (RIP2), a serine/threonine kinase, is an adaptor molecule of NOD1 and NOD2, and genetic variation in this receptor is known to be associated with the severity of allergic asthma in children. In this study, we examined the role of RIP2 in the development of allergic airway inflammation in a mouse model. METHODS: Airway inflammation was induced in mice through intranasal administration of ovalbumin (OVA) after 2 intraperitoneal immunizations with OVA. Lung inflammation and mucus hypersecretion were examined histologically and total cell infiltration in bronchoalveolar (BAL) fluids was determined. Levels of the Th2-related cytokines, IL-5 and IL-13, in lung extracts were measured by ELISA. Serum antigen-specific IgE and IgG1 levels were also assessed. RESULTS: OVA-induced lung inflammation and mucus hypersecretion were not different between WT and RIP2-deficient mice. The IL-5 and IL-13 levels in the bronchoalveolar (BAL) fluids were also not impaired in RIP2-deficient mice compared to WT mice. Moreover, RIP2 deficiency did not affect serum OVA-specific IgG1 and IgE levels. CONCLUSIONS: Our results suggest that RIP2 is not associated with the development of allergic airway inflammation.
Administration, Intranasal ; Airway Obstruction ; Animals ; Asthma ; Child ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Genetic Variation ; Humans ; Immunization ; Immunoglobulin E ; Immunoglobulin G ; Inflammation* ; Interleukin-13 ; Interleukin-5 ; Lung ; Methods ; Mice* ; Mucus ; Ovalbumin ; Ovum ; Phosphotransferases ; Pneumonia ; Receptors, Pattern Recognition

Asthma is characterized by chronic inflammation, goblet cell hyperplasia, the aberrant production of the Th2 cytokines, and eosinophil infiltration into the lungs. In this study, we examined the effects of baicalein, wogonin, and Scutellaria baicalensis ethanol extract on ovalbumin (OVA)-induced asthma by evaluating Th1/Th2 cytokine levels, histopathologic analysis, and compound 48/80-induced systemic anaphylaxis and mast cell activation, focusing on the histamine release from rat peritoneal mast cells. Baicalein, wogonin, and S. baicalensis ethanol extract also decreased the number of inflammatory cells especially eosinophils and downregulated peribronchial and perivascular inflammation in the lungs of mice challenged by OVA. Baicalein, wogonin, and S. baicalensis ethanol extract significantly reduced the levels of tumor necrosis factor α, interleukin (IL)-1β, IL-4, IL-5 and the production of OVA-specific IgE and IgG1, and upregulated the level of interferon-γ and OVA-specific IgG2a. In addition, oral administration of baicalein, wogonin, and S. baicalensis ethanol extract inhibited compound 48/80-induced systemic anaphylaxis and plasma histamine release in mice. Moreover, baicalein, wogonin, and S. baicalensis ethanol extract suppressed compound 48/80-induced mast cell degranulation and histamine release from rat peritoneal mast cells. Conclusively, baicalein and wogonin as major flavonoids of S. baicalensis may have therapeutic potential for allergic asthma through modulation of Th1/Th2 cytokine imbalance and histamine release from mast cells.
Administration, Oral ; Anaphylaxis* ; Animals ; Asthma ; Cytokines ; Eosinophils ; Ethanol* ; Flavonoids ; Goblet Cells ; Histamine Release* ; Histamine* ; Hyperplasia ; Immunoglobulin E ; Immunoglobulin G ; Inflammation* ; Interleukin-4 ; Interleukin-5 ; Interleukins ; Lung ; Mast Cells ; Mice ; Ovalbumin ; Ovum ; Plasma ; Rats ; Scutellaria baicalensis* ; Scutellaria* ; Tumor Necrosis Factor-alpha