The cytokine-driven regulation of secretoglobins in normal human upper airway and their expression, particularly that of uteroglobin-related protein 1, in chronic rhinosinusitis.

BACKGROUND: The involvement of secretoglobins (SCGBs) other than SCGB1A1 (Clara cell 10-kDa protein, CC10) in human airway diseases remains unexplored. Among those SCGBs, SCGB3A2 (uteroglobin-related protein 1, UGRP1) is particularly interesting, given its structure and function similarities with SCGB1A1 (CC10). The aim of this study was to investigate the expression regulation of SCGBs other than SCGB1A1 (CC10) in human upper airway, and their potential involvement, particularly that of SCGB3A2 (UGRP1), in chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) and without nasal polyps (CRSsNP). METHODS: Eight SCGB family members including SCGB3A2 (UGRP1), SCGB1C1 (ligand binding protein RYD5), SCGB1D1 (lipophilin A), SCGB1D2 (lipophilin B), SCGB1D4 (interferon-γ inducible SCGB), SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), and SCGB3A1 (uteroglobin-related protein 2) were studied. The regulation of SCGBs mRNA expression in normal nasal mucosa by proinflammatory, Th1, and Th2 cytokines was studied through nasal explant culture. SCGBs mRNA expression levels in CRSsNP and CRSwNP patients and controls were compared. The mRNA levels were detected by means of quantitative reverse transcriptase-polymerase chain reaction. The protein expression of SCGB3A2 (UGRP1) was analyzed using immunohistochemistry. RESULTS: The expression of SCGBs except SCGB1D2 (lipophilin B) could be found in upper airway and be differentially regulated by different cytokines. SCGB3A2 (UGRP1) mRNA expression was induced by Th1 cytokine, but suppressed by proinflammatory and Th2 cytokines. SCGBs mRNA expression was altered in CRS; particularly, SCGB3A2 (UGRP1) protein and mRNA expression was markedly decreased in both CRSsNP and CRSwNP and its protein levels inversely correlated with the number of total infiltrating cells, preoperative sinonasal CT scores, and postoperative endoscopy and symptom scores. CONCLUSION: SCGBs except SCGB1D2 (lipophilin B) are expressed in human upper airway and their expression can be differentially regulated by inflammatory cytokines. SCGBs mRNA expression is altered in CRS. Reduced production of UGRP1, which is likely due, at least in part, to a local cytokine environment, may contribute to the hyper-inflammation in CRS and correlates with response to surgery.

Increased serum complement component 3 and mannose-binding lectin levels in adult Chinese patients with chronic rhinosinusitis.

OBJECTIVE: To study the immune function of adult Chinese patients with chronic rhinosinusitis (CRS) to elucidate its potential role in the pathogenesis of CRS. METHODS: A prospective three-arm case-control study. The study population comprised 72 CRS patients without nasal polyps (NPs), 95 CRS patients with NPs, and 110 healthy controls. The concentrations of serum immunoglobulin A (IgA), M (IgM), G (IgG), IgG subclasses (IgG1-4), complement component 3 (C3), and complement component 4 (C4) were measured by nephelometry. Serum mannose-binding lectin (MBL) levels were analyzed by enzyme-linked immunosorbent assay. All CRS patients had a complete blood count with differential, atopic status evaluation, coronal computed tomographic (CT) scan of the sinuses, and nasal endoscopy. RESULTS: Frequency of immunoglobulin, C3, C4, or MBL deficiency showed no difference among groups. The prevalence of coexistence of MBL and immunoglobulin or complement component deficiency did not differ significantly among groups either. However, compared with controls, decreased IgG3 levels were found in CRS patients without NPs, and increased C3 and MBL levels was found in both CRS patients with and without NPs. Moreover, MBL levels were significantly higher in CRS patients with NPs than in CRS patients without NPs, which positively correlated with extent of disease seen on CT scan and endoscopy, and peripheral eosinophil count. CONCLUSIONS: Immunoglobulin, C3, C4, and MBL deficiency is not the main cause of CRS in adult Chinese patients. However, on the contrary, increased C3 and MBL levels in serum might play a modulatory role in CRS development.

Distinct immunopathologic characteristics of various types of chronic rhinosinusitis in adult Chinese.

BACKGROUND: Chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) and without nasal polyps (CRSsNP) is reported to be different in inflammatory patterns of the sinonasal mucosa in white patients. Studies in nonwhite populations may further be helpful to understand the pathogenic mechanisms of CRS. OBJECTIVE: To investigate the immunopathologic profiles of CRSwNP and CRSsNP in adult Chinese. METHODS: Histologic characteristics of surgical samples were analyzed in 50 controls, 94 CRSsNP patients, and 151 CRSwNP patients. Tissue samples from 17 controls, 36 CRSsNP patients, and 45 CRSwNP patients were stained for CD3, CD4, CD8, CD20, CD68, myeloperoxidase, and dendritic cell lysosome-associated membrane protein. Expression profiles of transcription factors of T-cell subsets in relation to cytokines and a marker of natural killer T cell (Valpha24) were examined by means of quantitative RT-PCR. RESULTS: Over half of CRSwNP patients presented noneosinophilic inflammation. CRSwNP had a higher number of eosinophils, plasma cells, and CD3(+), CD8(+), CD20(+), and CD68(+) cells and a lower myeloperoxidase expression rate than CRSsNP. Expression levels of transcription factors and cytokines of T(H)1/T(H)2/T(H)17 were increased, whereas the expression rate of Forkhead box p3 and TGF-beta1 was decreased in both CRSsNP and CRSwNP compared with controls. Comparing CRSsNP and CRSwNP, CRSsNP had higher levels of IFN-gamma expression, whereas only eosinophilic CRSwNP demonstrated an enhanced expression of GATA-3 and IL-5. Compared with noneosinophilic CRSwNP, an exaggerated T(H)2/T(H)17 reaction and Valpha24 expression were found in eosinophilic CRSwNP. CONCLUSION: Both Chinese CRSsNP and CRSwNP patients demonstrate impaired regulatory T cell function and enhanced T(H)1/T(H)2/T(H)17 responses. CRSsNP is confirmed to be a predominant T(H)1 milieu, whereas T(H)2 skewed inflammation with predominant T(H)17 reactions, and infiltration of natural killer T cells can be demonstrated only in eosinophilic CRSwNP, but not in noneosinophilic CRSwNP.

Lack of association of Clara cell 10-kDa protein gene variant with chronic rhinosinusitis in a Chinese Han population.

BACKGROUND: Clara cell 10-kDa protein (CC10) is an anti-inflammatory molecule and has been implicated in the involvement of the pathogenesis of asthma and chronic rhinosinusitis (CRS). A single nucleotide polymorphism (SNP) in CC10 gene (A + 38G) was previously shown to be associated with asthma and plasma CC10 levels. The purpose of this study is to examine whether there is an association between the CC10 A + 38G SNP, plasma CC10 levels, and CRS in a central Chinese population of Han nationality. METHODS: The CC10 A + 38G SNP was analyzed by means of polymerase chain reaction with restriction fragment length polymorphism and plasma CC10 levels were measured using enzyme-linked immunosorbent assay in 220 patients with CRS (90 patients with nasal polyps [NPs] and 130 patients without NPs) and 180 healthy control subjects. Among 220 patients with CRS, 108 patients were atopic subjects. Severity of disease was determined by coronal computed tomography (CT) scan in CRS patients, which was graded according to Lund and Mackay. RESULTS: The frequency of the A allele was 0.394, which was not significantly higher than the frequencies of other reported ethnic groups except for German. No association between the CC10 A + 38G SNP and CRS, any subgroup of CRS, or CRS severity could be found. Although subjects carrying the AA genotype had a significantly lower plasma CC10 concentration than those carrying the GG and GA genotypes in both CRS and control groups (p = 0.00 for all), no association was found between the plasma CC10 levels and CRS phenotype. CONCLUSION: The CC10 A + 38G SNP may not exert a substantial influence on the development of CRS in the Chinese Han population.

Histological and immunological observations of bacterial and allergic chronic rhinosinusitis in the mouse.

BACKGROUND: The purpose of this study was to elucidate histological and immunologic features of mouse models of bacterial chronic rhinosinusitis (BCRS) and allergic chronic rhinosinusitis (ACRS). METHODS: A BCRS mouse model was established using Streptococcus pneumoniae inoculation plus Merocel (Medtronic, Jacksonville, FL) ostiomeatal obstruction for 12 weeks. An ACRS mouse model was developed by means of ovalbumin (OVA) i.p. injection and subsequent repeated OVA intranasal challenge for 12 weeks. Histological changes of sinonasal mucosa of both models were examined by means of hematoxylin and eosin staining for general morphology and inflammatory cell, periodic acid-Schiff staining for goblet cell, and Masson-trichrome staining for collagen. Enzyme-linked immunosorbent assay was used to detect the concentrations of various cytokines in nasal lavage fluid. RESULTS: Polymorphonuclear neutrophil infiltration in lamina propria was more obvious in the BCRS model, whereas eosinophil infiltration was more apparent in the ACRS model. Significant goblet cell and subepithelial gland hyperplasia, subepithelial fibrosis, epithelial thickening, and mononuclear cell infiltration were shown in both models with more severe extent found in the ACRS model. Interleukin (IL)-6 and tumor necrosis factor alpha levels in NLF from both models were increased and peaked at 1 week. Interferon gamma levels were also up-regulated in both models but reached maximum at 1 week in the BCRS model and 4 weeks in the ACRS model. IL-8 (CXCL8) levels were only increased in BCRS mice and peaked at 1 week, whereas IL-5, IL-13, and eotaxin (CCL11) levels were only enhanced in ACRS mice and peaked at 1 week. The Th1/Th2 ratio in BCRS mice was significantly higher than that in ACRS mice (6.68 +/- 2.33 versus 1.37 +/- 0.86; p < 0.01). CONCLUSION: Histological and immunologic features of BCRS and ACRS mouse models were similar to those of human noneosinophilic and eosinophilic CRS, respectively. BCRS and ACRS mouse models have distinct immunologic characteristics and are applicable for CRS research.

[Expression difference of toll-like receptors among chronic rhinosinusitis and nasal polyps].

OBJECTIVE: To compare the expression difference of Toll-like receptor-2 (TLR-2) and Toll-like receptor-4 (TLR-4) protein and mRNA among the chronic rhinosinusitis tissues, nasal polyps tissues, and normal mucosa tissues. METHODS: The mRNA expression of TLR-2 and TLR-4 in chronic rhinosinusitis tissues from 10 patients, in nasal polyp tissues from 10 patients, and in inferior turbinate tissues from 10 patients underwent nasal septum operation was detected by means of reverse transcriptase-polymerase chain reaction (RT-PCR). Immunohistochemistry (IHC) was used to detected the expression of TLR-2 and TLR-4 protein in a different set of 20 chronic rhinosinusitis tissues, 20 nasal polyp tissues, and 20 normal inferior turbinate tissues. RESULTS: (1) The mRNA and protein expression of TLR-2 and TLR-4 was detected on epithelial and glandular cells membrane in all chronic rhinosinusitis, nasal polyps and control tissues . (2) The mRNA and protein expression of TLR-2 and the mRNA expression of TLR-4 in chronic rhinosinusitis tissues was significantly increased compared with that in nasal polyps and control tissues (P < 0.05). (3) The expression intensity of TLR-2 and TLR-4 mRNA and protein between nasal polyps and control tissues was found no significant difference (P > 0.05). CONCLUSIONS: Different TLR-2 and TLR-4 protein and mRNA level in chronic rhinosinusitis and nasal polyp tissues might imply that TLR-2 and TLR-4 play different role in the pathogenesis of chronic rhinosinusitis and nasal polyps.

[Relationship between the expression of tenascin C and TGF-beta1 in human nasal polyp tissues].

OBJECTIVE: To explore the mechanism underlying the increased expression of tenascin C (TNC) in human nasal polyp tissues. METHODS: The method of immunohistochemistry was used to detect the protein expression of TNC and transforming growth factor-beta1 (TGF-beta1) and their relationship in nasal polyp tissues. The cell culture, real-time RT-PCR and in situ ELISA techniques were employed to investigate the effect of TGF-beta1 and eosinophils on the TNC mRNA and protein expression in BEAS-2B immortalized human bronchial epithelial cells. RESULTS: (1) TNC and TGF-beta1 protein expression were up-regulated in nasal polyp tissues. TNC expression level was associated with the number of TGF-beta1 positive cells (r = -0.58, P < 0.01) and TGF-beta1 positive eosinophils (r = -0.61, P < 0.01); (2) 1 ng/ml and 10 ng/ml TGF-beta1 induced TNC mRNA expression by 7.20 +/- 3.43-fold and 22.48 +/- 5.35-fold (P < 0.01) in BEAS-2B cells after 4 h stimulation respectively. The fluorescence intensity of TNC protein expression was 129. 50 +/- 47.42 and 151.20 +/- 48.36 after 24 h stimulation respectively. The protein expression was significantly increased compared with that without stimulation (60.60 +/- 38.53, P < 0.05); (3) Coculture BEAS-2B cells and eosinophils at 2:1, 1:1 and 1:2 ratio, TNC mRNA expression was induced by 4.90 +/- 1.40-fold, 5.48 +/- 1.60-fold and 4.78 +/- 1.32-fold (P < 0.01) in BEAS-2B cells after 4 h coculture respectively. The fluorescence intensity of TNC protein expression was 128.75 +/- 44.15, 142.33 +/- 29.06 and 131.33 +/- 20.87 after 24 h coculture respectively. The protein expression was significantly increased compared with that without eosinophils coculture (59.40 +/- 10.14, P < 0.05). Treatment with neutralizing antibody to TGF-beta1 significantly inhibited eosinophil-induced BEAS-2B cells TNC expression in a dose-dependent manner (P < 0.05). CONCLUSION: TGF-beta1 and eosinophils can induce TNC expression in airway epithelial cells. The effect of eosinophils is partially mediated through TGF-beta1. Up-regulated expression of TNC in nasal polyp tissues is related to eosinophil-derived TGF-beta1.

[Cytogenetic aberrations of esthesioneuroblastoma studied by comparative genomic hybridization].

OBJECTIVE: To characterize the cytogenetic alterations of esthesioneuroblastoma (ENB). METHODS: Comparative genomic hybridization (CGH) was performed on genomic DNA extracted from 12 patients with primary ENB, 4 patients with tumor recurrence and 7 with metastasis. Equal amounts of biotin-labeled tumor DNA and digoxigenin-labeled normal reference DNA were hybridized to normal meta phase chromosomes. Tumor DNA was visualized by fluorescein (FITC) and normal DNA by rhodamin (TRITC ) and detected by fluorescence microscopy. The signal intensities of the different fluorochromes were quantitated as gray levels along the single chromosomes. The over-and under-represented DNA segments were determined by computation of FITC/TRITC ratio images and average ratio profiles. RESULTS: Consensus deletion regions were most frequently observed on chromosomes 1p, 2q, 3p/q, 4p/q, 5p/q, 6q, 8p/q, 9p, 10p/q, 11p, 12q, 13q, 18q, and 21q. DNA over-representations were identified on chromosomes 1p, 7q, 9q, 11q, 14q, 16p/q, 17p/q, 19p/q, 20p/q and 22p/q. The genetic pattern of ENB was distinct from that of other small round-cell tumor types and neuroblastomas. The deletion on chromosome band 1p21-p31 was associated with bad prognosis. In particular, all patients died whose tumors had combined 1p21-p31 deletion, with tumors in clinical stage C or D, and of low differentiation (grade III or IV). Clonality analysis revealed a high concordance between pairs of primaries and metastases. CONCLUSION: CGH analysis identifies characteristic cytogenetic aberrations of esthesioneuroblastoma associated with its malignant phenotype.

[Management of esthesioneuroblastoma of the Fulda surgical concept].

OBJECTIVE: To summaries the treatment strategy of esthesioneuroblastoma (ENB). METHODS: Between 1988 and 2001, 17 patients with ENB were treated at the Department of Otorhinolaryngology of the Klinikum Fulda. All patients were monitored on an outpatient basis after completed therapy with a median follow-up of 44 months. In a retrospective review, the patients' charts, the computed tomography, and magnetic resonance imaging scans, the operation reports, and the follow-up data were analyzed, particularly with respect to the surgical approaches. RESULTS: All tumors were staged according to Morita. One patient was classified as stage A, six stage B, nine stage C, and one stage D. All patients received surgical resection. Ten patients were disease free for at least 2 years, whereas 6 patients died because of ENB and one due to other disease. Of 10 patients who were free of disease, the tumors were removed via a transnasal approach in 6 patients using the microscope in combination with the endoscope. These tumors resected endonasally were staged as A (1 case) and B (5 cases). In ENB of stage C a craniofacial resection was performed using a subfrontal approach or the midfacial degloving. The lateral rhinotomy was applied only in cases in which an exenteration orbitae had to be carried out. CONCLUSION: ENB is best managed by complete surgical resection followed by adjuvant stereotactic radiation therapy. The Fulda surgical concept in management of anterior skull base tumors is also forwarded to resection of ENB. It classifies the following indications: 1) Endonasal approach in cases without tumor infiltration of the orbit and/or the brain; 2) Subfrontal approach in cases with extended tumor infiltration of the intradural space or the brain; 3) Midfacial degloving in cases with far lateral tumor spread, particularly fossa pterygoidea or pterygopalatina; 4) Lateral rhinotomy in all cases where an exenterative orbita is needed.