Antigens, Bacterial ; Humans ; Inflammation ; Nasal Mucosa ; immunology ; Staphylococcus ; immunology ; Superantigens ; immunology

OBJECTIVE: The aim of this study was to use a guinea pig model of prolonged allergic-induced rhinitis to characterize the feature of nasal mucosa remodeling. METHOD: Forty-eight male Hartley guinea pigs were randomly divided into six groups: allergen challenged groups (Group OVA(2w) , Group OVA(6w) and Group OVA(12w)) and control groups respectively (Group Sal(2w), Group Sal(6w) and Group Sal(12w)). Each group had 8 guinea pigs. To develop a guinea pig model of nasal mucosa remodeling, ovalbumin-sensitized guinea pigs were repeatedly challenged with allergen twice a week from two weeks to 12 weeks. Matched control groups were challenged with physiological saline. Nasal lavage was performed 24 hours after the last intranasal challenge. Then nasal mucosa were obtained. HE, AB-PAS, MT, and immunohistochemical staining against transforming growth factor-beta1 (TGF-beta1) were performed. Eosinophil cationic protein (ECP) and OVA-special IgE (OVA-sIgE) were detected by ELISA in nasal lavage fluid. RESULT: (1) The levels of OVA-sIgE in nasal lavage fluid in Group OVA(6w) and Group OVA(12w) were significantly different from Group OVA(2w), while the levels of ECP had no significant difference among the experiment groups. The levels of OVA-sIgE and ECP in experiment groups were significantly different from control groups respectively (P < 0.01). (2) Grade 0 and Grade 1 of epithelial damage were significantly different in Group OVA(6w) and Group OVA(12w) when compared with from Group OVA(2w) (P < 0.01). At the same time, Grade 0 and Grade 1 of epithelial damage were statistically different in the experiment groups when compared with the respectively control groups (P < 0.05). (3) Goblet gland hyperplasia and collagen deposit within the extracellular matrix (ECM) were easily found in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). The number of goblet gland and the ratio of collagen deposit were statistically more in Group OVA(6w) and Group OVA(12w) than in Group Sal(6w) and Group Sal(12w) (P < 0.05). That feature of the ratio of collagen deposit did not show in Group OVA(2w) versus Group Sal(2w). (4) Increased TGF-beta1 expressions were observed in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). Those increasing expressions were also observed in experiment groups rather than in the respectively control groups (P < 0.01). CONCLUSION Epithelial damage, goblet cells hyperplasia and collagen deposition in ECM were observed as the features of remodeling in this guinea pig model of allergic rhinitis under prolonged allergen challenge. Epithelial damage, excessive expression of related cytokines and enhancement activity of enzymes were observed in early time after challenge of allergen. The features of goblet cells hyperplasia and collagen deposition in ECM were observed at a later stage.
Airway Remodeling ; Allergens ; immunology ; Animals ; Disease Models, Animal ; Guinea Pigs ; Male ; Nasal Mucosa ; immunology ; pathology ; Rhinitis, Allergic, Perennial ; immunology ; pathology

BACKGROUNDExcessive expression of thymic stromal lymphopoietin (TSLP) has been demonstrated in asthmatic airway epithelia and in nasal epithelia from animal models of allergic rhinitis (AR), but the evidence of expression of TSLP in nasal epithelial cells (NECs) of patients with AR is lacking. We aimed to investigate the expression of TSLP in NECs of patients with mugwort sensitive-seasonal AR and determine whether it is associated with severity of symptoms and the number of infiltrated eosinophils in nasal mucosa.

METHODSNECs specimens were obtained by scraping with plastic curettes from the nasal inferior turbinates of patients with mugwort pollen sensitive-seasonal AR (n = 22) and nonallergic controls (n = 11) during last peak mugwort pollen season. The severity of nasal symptom was assessed using a Visual Analog Scale (VAS). In addition, serum mugwort pollen IgE levels were tested from each patient. In situ hybridization (ISH) was performed to test the messenger RNA (mRNA) of TSLP in the NECs. Furthermore, immunohistochemical staining (IHC) was scored to evaluate the expression of TSLP and eosinophil cell count was made by May-Grünwald/Giemsa staining. The correlation between expression of TSLP and all other parameters was analyzed in this study.

RESULTSThe mRNA level of TSLP was significantly increased in NECs of patients with AR compared with the nonallergic control group (P < 0.05). In addition, IHC results showed that expression of TSLP in NECs from patients with AR was up-regulated which was correlated with VAS score (r = 0.598; P < 0.05) and nasal eosinophils count (r = 0.702; P < 0.05), but it was unrelated with mugwort pollen specific IgE level.

CONCLUSIONSThese preliminary findings indicate a potential relationship between TSLP expression, severity of symptoms and nasal eosinophils count in pathogenesis of AR, but TSLP expression did not correlate with mugwort pollen specific IgE level. The elevated expression of TSLP might play a critical role in local atopical responses of AR. In the future, the TSLP has the potential to be one of the most important molecular markers for AR diagnoses and assessment.

Adolescent ; Adult ; Artemisia ; immunology ; Cytokines ; analysis ; genetics ; Female ; Humans ; Male ; Middle Aged ; Nasal Mucosa ; immunology ; Pain Measurement ; Pollen ; immunology ; RNA, Messenger ; analysis ; Rhinitis, Allergic, Seasonal ; immunology

OBJECTIVE: To investigate the expression of interleukin (IL)-23 in the nasal mucosa of allergic rhinitis patients and its significance. METHOD: mRNA and protein expression of IL-23 in inferior turbinate mucosa from 12 allergic rhinitis patients and 11 control patients was measured by means of real-time RT-PCR and immunohistochemistry, respectively. RESULT: IL-23p19 mRNA relative expression level in nasal mucosa was significantly increased in allergic rhinitis patients compared with normal controls (P < 0.01). Immunohistochemical staining demonstrated that IL-23 protein was mainly expressed by infiltrating inflammatory cells in lamina propria and there was increased number of IL-23 positive cells in allergic rhinitis patients in comparison with normal controls. Correlation analysis showed that the mRNA and protein expression level of IL-23 was significantly positively correlated with the number of the inflammatory cells (r = 0.678 and 0.644, respectively; both P < 0.01) and the degree of subepithelial collagen deposition (r = 0.834 and 0.721, respectively; both P < 0.01). IL-23p19 mRNA relative expression level in nasal mucosa was significantly decreased in allergic rhinitis patients who used glucocorticoids compared with controls (P < 0.01). CONCLUSION IL-23 may contribute to the chronic inflammation and airway remodelling in allergic rhinitis.
Adolescent ; Adult ; Case-Control Studies ; Female ; Humans ; Inflammation ; immunology ; Interleukin-23 ; immunology ; Male ; Middle Aged ; Nasal Mucosa ; immunology ; metabolism ; Rhinitis, Allergic, Perennial ; immunology ; Young Adult

OBJECTIVE: To explore the role of the innate immune factors TLR2 and TLR4 in the pathogenesis of chronic rhinosinusitis (CRS) by detecting their expression in different clinical types of CRS and the normal control group. METHOD: Immunohistochemistry was used to detect the expression of TLR2 and TLR4 respectively in 21 cases (chronic rhinosinusitis with nasal polyps, CRSwNP) group, 15 cases (chronic rhinosinusitis without nasal polyos, CRSsNP) group, 11 cases recurrent CRSwNP group and 13 cases control group. Positive cells were counted under the microscope artificially, Mann-Whitney U analysis was applied for the ranked data, and one-way anova analysis was adopted to analyze the experimental group and control group. RESULT: (1) TLR2 and TLR4 expression had the same characteristics. Expression mainly concentrated in parts of the whole layer of epithelial basement membrane, cytoplasm of glandular cells, very few inflammatory cells such as monocytes and plasma cells in the cytoplasm, sometimes unknown cell nuclei positive expression. (2) The glandular cells were stained manual counting and color grading. TLR2 and TLR4 packet application Wilcoxon rank test Mann-Whitney U test analysis was not statistically significant (P > 0.05), measurement data within the group variance statistical difference between the groups (P < 0.05). CONCLUSION The Nasal mucosa can produce the innate immune factors TLR2 and TLR4. The different expression of TLR2 and TLR4 in the various clinical types of CRS suggests that they play the certain role in the pathogenesis of CRS.
Chronic Disease ; Epithelial Cells ; immunology ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Nasal Mucosa ; immunology ; metabolism ; Nasal Polyps ; immunology ; metabolism ; Rhinitis ; immunology ; metabolism ; Sinusitis ; immunology ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo construct a replication-defective recombinant adenovirus expressing the ORF2 (112-660aa) antigen of hepatitis E virus (HEV) and evaluate its immunization effect in BALB/c mice by mucosal inoculation.

METHODSThe HEV ORF2 gene encoding for 112-660aa was amplified from plasmid pUC-HEV and inserted into the transfer vector pTrack-CMV. The recombinant plasmid and adenoviral backbone plasmid pAdEasy-1 were co-transformed into E. coli strain BJ5183. Taking the advantage of the high efficient homologous recombination machinery presented in bacteria, the recombinant adenovirus backbone plasmid was generated in BJ5183, and then was transfected into 293 cells. Recombinant Adenoviruses were propagated in 293 cells with high titers. 8-week-old BALB/c mice were inoculated intraperitoneally and intranasally with 10(7) pfu recombinant adenovirus each on weeks 0, 3, 5, 7, 10.

RESULTSBoth groups of mice induced humoral IgG immune response with the highest titers 1:1,000 and 1:10,000 each. Only the group inoculated intranasally could induce mucosal IgA immune response.

CONCLUSIONSThe adenoviral recombinant can stimulate specific humoral and mucosal immune response in mice and is potentially to be used as a candidate vaccine for the treatment of HEV infection.

Adenoviruses, Human ; genetics ; Animals ; Hepatitis Antigens ; genetics ; immunology ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; immunology ; Peritoneum ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Viral Hepatitis Vaccines ; Viral Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo investigate the effect of early allergen exposure on later development of allergic rhinitis in mouse.

METHODSTwenty-four BALB/c neonates were randomly divided into 4 groups (low-dose group, high-dose group, negative control group and positive control group), each group had 6 mice. The mice were administered ovalbumin (OVA) by subcutaneous injection on day 1, 5, 12 after birth (10 μg OVA in 0.05 ml saline for low-dose group, 1000 μg OVA in 0.05 ml saline for high-dose group, only saline for negative and positive control group). Then the mice were sensitized and intranasally challenged with OVA (saline without OVA was used in negative control group) after 6 weeks. Symptoms, histopathological changes of nasal mucosa were observed, OVA-IgE in serum was examined, cytokines IL-4, IL-5 and IFN-gamma were detected in the supernatant of cultured splenic mononuclear cells.

RESULTSCompared to the positive control group, symptoms and nasal mucosa histological changes of high-dose group was indistinctive. The level of OVA-IgE and cytokines IL-4, IL-5 (x(-) +/- s) in high-dose group [(265.11 +/- 26.29), (446.39 +/- 72.83) and (171.24 +/- 15.66) pg/ml, respectively] were significantly lower than those in positive control group [(665.85 +/- 43.15), (1113.45 +/- 30.47), (255.36 +/- 30.96) pg/ml, respectively, t value were 0.000, 0.000 and 0.009, respectively, all P < 0.05]. The level of IFN-γ in high-dose group [(319.74 +/- 56.30) pg/ml] was significantly higher than those in positive control group [(170.02 +/- 14.50) pg/ml, t = 0.000, P < 0.05]. There was no significant difference of the results between the low-dose group and positive control group.

CONCLUSIONSNeonatal immunization with high-dose OVA inhibited the future allergic rhinitis symptoms, nasal histological changes, serum OVA-IgE levels and Th1/Th2 cytokine imbalance, resulting in the protective effect.

Allergens ; immunology ; Animals ; Animals, Newborn ; Dose-Response Relationship, Immunologic ; Immunoglobulin E ; blood ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Interleukin-5 ; analysis ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; immunology ; Nasal Provocation Tests ; Ovalbumin ; immunology ; Rhinitis, Allergic, Perennial ; immunology ; prevention & control

OBJECTIVE: To investigate the expression and role of Interleukin-33 (IL-33) and ST2 in the nasal polyps of human Eosinophilic and non-Eosinophilic chronic rhinosinusitis with nasal polyps (ECRS and non-ECRS). METHOD: IL-33 and ST2 protein expression in nasal polyps of ECRS and non-ECRS as well as in seemingly normal mucosa of the inferior turbinate tissue was investigated by immunohistochemical staining and messenger RNA (mRNA) expression of IL-33 and ST2 was assessed by realtime polymerase chain reaction (PCR) in 27 subjects with ECRS, 33 subjects with non-ECRS, and 11 control subjects. RESULT: (1) The ST2 was found both in nasal polyps of ECRS and non-ECRS,especially in ECRS, yet hardly found in the normal mucosa of the inferior turbinate tissue; (2) The expression of ST2 mRNA in nasal polyps of ECRS was higher than that in non-ECRS and normal inferior turbinate tissue, and the difference was both prominent in statistics (P<0.01); (3) The expression patterns of IL-33 at both mRNA and protein levels were not significantly different among the three groups (P>0.05). CONCLUSION The IL-33 and its receptor ST2 were both expressed in human nasal polyps including ECRS and non-ECRS, meanwhile the expression patterns of ST2 at both mRNA and protein levels were significantly higher in nasal polyps of ECRS. The current study suggests that IL-33 and its receptor ST2 may play important roles in the pathogenesis of chronic rhinosinusitis with nasal polyps, especially in ECRS through the increased expression of ST2 in Eosinophils as a hypothesis.
Chronic Disease ; Eosinophils ; immunology ; Humans ; Interleukin-1 Receptor-Like 1 Protein ; Interleukin-33 ; metabolism ; Nasal Mucosa ; metabolism ; Nasal Polyps ; immunology ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Receptors, Cell Surface ; metabolism ; Rhinitis ; immunology ; Sinusitis ; immunology ; Turbinates ; metabolism

OBJECTIVE: To investigate the expression of CD4, CD69, CD34, RANTES, IL-5 and IL-8 in nasal polyp tissues, and study their roles in the formation of nasal polyp. METHOD: The expression of CD4, CD69, CD34, RANTES, IL-5 and IL-8 were detected by immunohistochemical method and image analysis in 34 cases of nasal polyps and 30 cases of nasal concha mucosa (LNT). RESULT: The positive rate of glandular organ hyperplasia, formation of beaker cell, fiber hyperplasia, interstitial edema and infiltration of lymphocyte and eosinophilic granulocyte in nasal polyps were significantly higher than those in nasal concha mucosa (P<0.01). The cell density (piece/mm2) of CD4+, CD69+, IL-5, IL-8, RANTES in 34 nasal polyps was significantly higher than those in nasal concha mucosa (P<0.05). Marked positive correlations were found between expression of CD4, CD69 and RANTES, IL-5 and IL-8 (P<0.05, P<0.01 and P<0.05), expression of IL-5 and RANTES and infiltration level of eosinophilic granulocyte (P<0.05 and P<0.01), and expression of IL-8 and vaso formation on nasal polyps tissue (P<0.01). CONCLUSION T lymphocytes and correlated cytokines participate in the immunopathogenesis of nasal polyps; IL-5 and RANTES can prompt the infiltration, the aggregation and the activation of eosinophilic granulocytes; IL-8 can promote the vaso formation in nasal polyps.
Chemokine CCL5 ; metabolism ; Female ; Granulocytes ; immunology ; Humans ; Interleukin-5 ; metabolism ; Interleukin-8 ; metabolism ; Lymphocyte Activation ; Lymphocyte Count ; Male ; Middle Aged ; Nasal Mucosa ; immunology ; metabolism ; Nasal Polyps ; immunology ; metabolism ; T-Lymphocytes ; immunology

OBJECTIVESTo study the concentration, distribution and expression of IL-5 in nasal polyp tissues and explore its significance in micro-environment differentiation of eosinophil accumulation.

METHODSThe concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers were used as control.

RESULTSIL-5 concentration in polyp tissues was significantly higher than that in turbinate mucosa (P < 0.05). There was no significant difference in the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentrations in polyp tissues were markedly higher in patients with allergic rhinitis compared with those without (P < 0.05). IL-5 concentrations had no correlation with age and sex (P > 0.05). 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of lymphocytes and neutrophils were positive for IL-5; IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05); there was no significant difference in the the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissue was significantly stronger in patients with allergic rhinitis compared with those without (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and turbinate nasal mucosa (both P > 0.05).

CONCLUSIONIL-5 is the key cytokine in eosinophilic pathologic mechanisms in nasal polyp tissues.

Adult ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; chemistry ; Female ; Humans ; Interleukin-5 ; analysis ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Nasal Mucosa ; chemistry ; Nasal Polyps ; immunology ; Turbinates ; chemistry