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

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

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 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 observe expression and distribution of histamine H4 receptor in nasal mucosa in normal people and allergic rhinitis patients,and understand role of histamine H4 receptor in allergic rhinitis. METHOD: Select normal people and allergic rhinitis patients each 10, take the nasal mucosa, detect expression and distribution of histamine H4 receptor at proteins and transcription level respectively by immunohistochemical method and RT-PCR, and compared. RESULT: Histamine H4 receptor at proteins and transcription level were found in normal nasal mucosa (25 509 +/- 6 441, 0.42 +/- 0.08), increased significantly in nasal mucosa of allergic rhinitis patients (49 676 +/- 8 541, 0.69 +/- 0.11, P < 0.05), which in structural cells and immune cells. CONCLUSION Histamine H4 receptors exist in normal nasal mucosa, its express significantly enhance, flew histamine H4 receptor may be mediated histamine in pathogenesis of allergic rhinitis ,who is one of the ligands of histamine.
Case-Control Studies ; Humans ; Nasal Mucosa ; immunology ; metabolism ; pathology ; Receptors, G-Protein-Coupled ; metabolism ; Receptors, Histamine ; metabolism ; Receptors, Histamine H4 ; Rhinitis, Allergic, Perennial ; immunology ; metabolism ; pathology

OBJECTIVE: The aim of this study was to explore the changes of the extracellular matrix in nasal mucosa by a guinea pig model of prolonged allergic-induced rhinitis. METHOD: Thirty-two male Hartley guinea pigs were randomly divided into four groups: allergen challenged groups (Group 2 w, Group 6 w and Group 12 w) and a control group. Ovalbumin-sensitized guinea pigs were repeatedly challenged with allergen twice a week from 2 weeks to 12 weeks. Matched control groups were challenged with physiological saline. Nasal mucosa were obtained from the animals killed. Hematoxylin-Eosin, Masson's trichrome, and immunohistochemical staining against transforming growth factor-beta1 (TGF-beta1), Collagen III and Collagen I were performed to nasal mucosa. RESULT: (1) Pathological examination showed obvious infiltration of eosinophils and the enlarged thickness of epithelial layer of nasal mucosa in the experiment groups. (2) The area ratios of blue stained in the extracellular matrix of nasal mucosa were increased. The area ratios of blue stained were statistically different in Group 6 w and Group 12 w compared with the control group. (3) The increasing absorbance of TGF-beta1 were statistically different in the experiment groups with the control group. The absorbance of Collagen III and Collagen I showed a rising trend along prolonged allergen challenged in the experiment groups. CONCLUSION Prolonged allergen challenge and the inflammation of nasal mucosa, can lead to the increasing of the inflammation relevant factors and the deposit of collagen in the extracellular matrix of nasal mucosa.
Allergens ; immunology ; Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Eosinophils ; immunology ; Extracellular Matrix ; immunology ; metabolism ; pathology ; Guinea Pigs ; Inflammation ; Male ; Nasal Mucosa ; immunology ; metabolism ; pathology ; Rhinitis, Allergic, Perennial ; immunology ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVE: To determine the presence of TLR-9 protein in primary epithelial cell cultures of chronic rhinosinusitis with nasal polyps (CRSwNP), and then explore the role of innate immune recognition in the pathogenesis of CRSwNP. METHOD: Primary epithelial cell cultures were established in 10 controls and 10 CRSwNP patients who underwent sinus surgery, and flow cytometry was used to confirm its purity and measure the expression of TLR-9 protein. RESULT: By digestive method 98% of cultured primary nasal mucosa cells were epithelial cells and the expression of TLR-9 protein in CRSwNP group (11%-15%) was lower than that in normal group (49%-60%). CONCLUSION This finding suggests that impaired innate immune responses via TLR-9 on sinonasal epithelial cells may involve in pathogenesis of CRSwNP.
Adolescent ; Adult ; Aged ; Cells, Cultured ; Chronic Disease ; Epithelial Cells ; metabolism ; Female ; Humans ; Immunity, Innate ; Male ; Middle Aged ; Nasal Mucosa ; immunology ; metabolism ; pathology ; Nasal Polyps ; immunology ; metabolism ; Sinusitis ; immunology ; metabolism ; Toll-Like Receptor 9 ; metabolism ; Young Adult

OBJECTIVETo explore a new immunotherapy against allergic rhinitis.

METHODSThe recombinant protein of CTLA4 extracellular domain was obtained through construction of CTLA4-yeast expression system. The allergic rhinitis in mice was induced by sensitizing and challenging with ovalbumin (OVA). The allergic rhinitis related symptoms and the morphological changes in nasal mucosa were compared between the allergic rhinitis group and the CTLA4 extracellular domain group treated with CTLA4 extracellular domain before each challenge by ways of intraperitoneal injection.

RESULTSCTLA4 extracellular domain with a molecular weight of 28 000, which was confirmed by Western blot, could be generated through CTLA4-yeast expression system. The purified CTLA4 extracellular domain could inhibit T cells proliferation in mixed lymphocyte reaction with a inhibitory rate of 95.4%. The mice in allergic rhinitis group appeared typical allergic rhinitis symptoms after OVA challenge, such as rhinorrhea and sneeze. Meanwhile the nasal pathological studies showed edema and congestion in mucosa tissue and local influx of inflammatory cells. Whereas in CTLA4 extracellular domain group, the nasal symptoms were rarely observed, and the pathological change in nasal mucosa was significantly abated.

CONCLUSIONSThe protein of CTLA4 extracellular domain could prevent the allergic rhinitis in mice. The underlying mechanism of which might be the inhibition of the T cell activation.

Animals ; Antigens, CD ; immunology ; metabolism ; pharmacology ; CTLA-4 Antigen ; Cells, Cultured ; Humans ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; pathology ; Ovalbumin ; immunology ; metabolism ; Recombinant Proteins ; immunology ; metabolism ; pharmacology ; Rhinitis, Allergic, Perennial ; immunology ; pathology ; prevention & control

OBJECTIVETo evaluate the therapeutic effect of intranasal oligodeoxynucleotides with CpG motifs (CpG ODN) in prevention of allergic rhinitis in juvenile guinea pigs.

METHODSJuvenile guinea pigs aged from 7 to 10 weeks were administrated with CpG ODN alone or combined with OVA at single dose concentration intranasally (on day 0, 5, 10, 15 in sequence) while control and blank group were administrated with saline. Both experimental and control animals were again sensitized by OVA (on day 18, 25), and 14 days after second sensitization animals were challenged by OVA intranasally (on days 39 and 46). Two hours after challenge, the animals were sacrificed. Then Hemotoxin and Eosin stain were carried out to analyze local eosinophilic reactions and nasal lesions. Local and systemic cytokines interleukin IL-5 and IFN-γ levels were examined by ELISA. Immunofluorescence was carried out with ICAM-1 antibody. Statistical analysis was performed using a SPSS 11.0 software.

RESULTSIn CpG ODN-administration or CpG ODN with OVA-administration group allergic rhinitis symptoms were not as severe as model control group (P < 0.05). Compared with the model control group, CpG ODN-administration did not increase production of OVA-specific Th1 cytokine IFN-γ but decreased productions of ovalbumin-specific Th2 cytokines IL-5 both in serum and nasal specimen (q value were 3.890 and 4.019, P < 0.05). Moreover, nasal lesions with infiltration of mean (x ± s) eosinophils (20.0 ± 9.6) in CpG group animal were prominently reduced by the CpG ODN-treatment compared with the control animals (53.5 ± 19.8) and CpG+OVA group (9.5 ± 5.7) were lower than CpG-M+OVA group (49.2 ± 18.9), the differences were significant (q value were 3.785 and 4.576, P < 0.05). Immunofluorescence results showed lower ICAM-1 expression in nasal specimen of CpG group compared with model group and CpG plus OVA group animal to CpG mimics plus OVA group (Z value were 3.697 and 3.765, P < 0.05).

CONCLUSIONSIntranasal administration of CpG oligodeoxynucleotides with or without allergen may be an effective way to prevent the development of allergic rhinitis.

Animals ; Disease Models, Animal ; Guinea Pigs ; Intercellular Adhesion Molecule-1 ; metabolism ; Interferon-gamma ; immunology ; Interleukin-5 ; immunology ; Male ; Nasal Mucosa ; pathology ; Oligodeoxyribonucleotides ; immunology ; therapeutic use ; Rhinitis, Allergic, Perennial ; immunology ; prevention & control ; Toll-Like Receptor 9 ; immunology ; therapeutic use

OBJECTIVE: To explore the changes of microRNAs in nasal mucosa after the specific immunotherapy (SIT) for allergic rhinitis (AR) in mice. METHOD: Female BALB/c mice, 6-8 weeks of age, were randomly divided into control group, model group and treatment group. AR model were established by intraperitoneal injection and intranasal challenge of ovalbumin and SIT was performed by inguinal subcutaneous injections. AR symptom scores were documented. The eosinophils (EOS) in the nasal mucosa were measured. Ovalbumin-specific IgE (OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calculated. The microRNAs in the nasal mucosa were preliminary screened by microRNA gene microarray. Comparing with model group, the Fold changes of microRNA of the treatment group were ≥ 2.0 and the P < 0.05. MicroRNA target genes were predicted with GeneSpring 12.5 software. We took the intersection between genes in the signal pathway which associated with immune response,inflammation and target genes. The MEV-4-6-0 and Cytoscape_v2. 8. 2. software was applied to perform the cluster analysis and target gene regulatory networks maps. RESULT: The model of AR in mice and its SIT were successful. Comparing with the model group, the Fold changes of 15 microRNAs, of which 9 microRNAs were up-regulated and 6 microRNAs were down-regulated, were ≥ 2.0 in treatment group (P < 0.05). Cluste analysis showed clearly that microRNAs in the treatment group and model group respectively aggregated in two branches. The 15 microRNAs had 5302 target genes, of which, 451 genes were related more with SIT by the intersection. One microRNA can regulate many target genes, and one gene can also be affected by many microRNAs. Their synergistic effects may be involved in the mechanism of SIT. CONCLUSION The expressions of microRNAs are changed in nasal mucosa after SIT for AR in mice and we can speculate that microRNAs are involved in the process of SIT for AR. Bioinformatics methods can diminish the scope of target genes of microRNAs, which will help us studying the effect of changed microRNA on its relative target genes after SIT, and make us better understanding the mechanism of the disease and its SIT.
Administration, Intranasal ; Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; immunology ; Female ; Immunoglobulin E ; blood ; Immunotherapy ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Mice ; Mice, Inbred BALB C ; MicroRNAs ; metabolism ; Nasal Mucosa ; drug effects ; metabolism ; Ovalbumin ; Rhinitis, Allergic ; therapy