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

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*

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

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*

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

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

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

OBJECTIVES: Lidocaine, a local anaesthetic is a treatment option in uncontrolled asthma due to its immunomodulatory effects. In the present study, proparacaine (PPC), a derivative of lidocaine was examined for its therapeutic application in a mouse model of allergic rhinitis. METHODS: The mice were grouped into 4 groups: control group, allergic rhinitis (AR) group, ciclesonide (CIC) group, and PPC group. Nasal symptom scores, eosinophil counts, goblet cell counts, and mast cells counts in the nasal mucosa were measured. Serum ovalbumin (OVA)-specific immunoglobulin (Ig) E, OVA-specific IgG1, OVA-specific IgG2a, interleukin (IL)-4, IL-5, and cortisol levels were measured. RESULTS: Intranasal administration of PPC significantly decreased nasal symptoms, number of eosinophils, goblet cells, and mast cells in the lamina propria of the nasal mucosa. Serum OVA-specific IgE, OVA-specific IgG1, OVA-specific IgG2a was significantly higher in the AR compared with the control group. Serum level of IL-4 was significantly lower in the CIC group and PPC group in comparison with AR group. Serum IL-5 showed no significant difference among all groups. No significant difference in serum cortisol levels was observed among the 4 groups. CONCLUSION: PPC appears to have a therapeutic potential in treatment of allergic rhinitis in a mouse model by reducing eosinophil, goblet cell, and mast cell infiltration in the nasal mucosa.
Administration, Intranasal ; Animals ; Asthma ; Eosinophils ; Goblet Cells ; Hydrocortisone ; Immunoglobulin E ; Immunoglobulin G ; Immunoglobulins ; Interleukin-4 ; Interleukin-5 ; Interleukins ; Lidocaine ; Mast Cells ; Mice* ; Mucous Membrane ; Nasal Mucosa ; Ovalbumin ; Rhinitis, Allergic*

OBJECTIVETo observe the effect of Yupingfeng San (YPFS) against OVA-induced allergic asthma in mice.

METHODMice were injected with OVA to establish the allergic asthma model. They were abdominally injected with 20 microg OVA on day 0 and 14, and inhaled aerosol 0.5% OVA solution for 20 min for seven days. The blank control group was administrated with equal volume of saline. YPFS groups with different doses were administrated intragastrically with YPFS every day, with the crude drug dosage of 3.25, 6.5, 13 g x kg(-1), respectively. The model group and control group were administrated with equal volume of saline. The positive control group was given intraperitoneally injected with 1 mg x kg(-1) DEX since aerosol inhalation. Blood was drawn after the last OVA aerosol inhalation to count the number of Eosnophils (Eos) in blood and detect IgE in serum; BALF was collected to count the number of cells and classify; right lung tissues were evenly grinded to detect cytokines IL-4 and IFN-gamma, and left upper lung lobes were collected for pathologic histology.

RESULTThe level of Eos and IgE in serum increased significantly in the model group, and a large number of Eos were detected in BALF. Histopathological changes in lung showed bronchial serous exudation, tubular epithelial cells exfoliation, tube narrowing, widened alveolar septum, and bronchial periarterial lymphocytes infiltration. Homogenate of lung tissues showed increase of IL-4, and decrease in IFN-gamma/IL-4 ratio. YPFS groups with different doses displayed decrease of Eos in blood and BALF and IgE content in serum, and relief of pathologic changes in above models. Meanwhile, IL-4 content in homogenate of lung tissues decreased, with the increase in IFN-gamma/IL-4 ratio.

CONCLUSIONYPFS shows the inhibitory effects on OVA-induced allergic asthma, involving down regulation of Eos and IgE levels in blood of asthma mice, and infiltration of inflammatory cells in lung tissues. Meanwhile, it can reduce IL-4 in lung homogenates, increase IFN-gamma/IL-4, and inhibits Th2 polarization.

Animals ; Asthma ; chemically induced ; drug therapy ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Eosinophils ; drug effects ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; adverse effects

OBJECTIVEThe incidence of food allergy has increased in recent years and there is no effective way to treat it except strict dietary avoidance and rapid medical treatment in case of accidental exposure. Oral tolerance, as a new method, has shown great promise as an alternative approach to prevention and treatment for allergic disease. It was reported that all-trans retinoic acid (atRA) plays an important role in inducing oral tolerance in vitro. Our study aimed to investigate the immunological effect of different doses of atRA on ovalbumin (OVA) allergic BALB/c mice.

METHODBALB/c mice were sensitized by intraperitoneal injection with OVA to establish allergic animal model. According to the dose of atRA given, 40 OVA allergic BALB/c mice were divided into 4 groups: the mice in high dose group were treated with 100 mg/kg atRA (atRA-H), those in median dose group were treated with 50 mg/kg atRA (atRA-M), those in low dose group were treated with 20 mg/kg atRA (atRA-L) and the mice in control group were given vehicle-soy oil only (CTR). After 12 days of atRA intervention, weight was measured, the mice were checked for diarrhea , and intestinal histology was observed after hematoxylin and eosin staining. The level of OVA-IgE in serum, total IgA and OVA-IgA in feces were measured by ELISA. The percentage of CD4⁺ CD25⁺ FoxP3⁺ T cells in CD4⁺ T cells in mesenteric lymph node was detected by flow cytometry.

RESULTCompared with that of CTR group, the level of OVA-IgE in serum (1.221 ± 0.367 vs. 0.793 ± 0.616) and OVA-IgA (1.573 ± 0.656 vs. 0.905 ± 0.279) in feces decreased significantly (P = 0.006 and 0.012, respectively) without weight and intestinal histology changes after low dose of atRA administration. However, there was no significant difference in the percentage of CD4⁺ CD25⁺ FoxP3⁺ T cells in CD4⁺ T cells in mesenteric lymph node (10.641 ± 1.218 vs. 10.936 ± 0.954) between atRA-L and CTR group (P > 0.05). While in animals with high and median dose of atRA administration, no immunologic improvement was found, instead, there was weight loss and intestinal mucosal damage.

CONCLUSIONLow dose of atRA intervention seems to induce immune suppression in vivo resulting in positive effects on OVA allergic mice. However, median and high dose atRA had no therapeutic effect on OVA allergic mice.

Animals ; CD4-Positive T-Lymphocytes ; Food Hypersensitivity ; drug therapy ; immunology ; Immune Tolerance ; drug effects ; Lymph Nodes ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; T-Lymphocytes, Regulatory ; Tretinoin ; administration & dosage ; pharmacology