Search

Background: Nasal polyposis is mostly associated with eosinophilia of mucosal tissue. This points to the implication of CC chemokines in nasal eosinophilia. Recently the CC chemokine eotaxin-2 (CCL24) was identified. This study was initiated to localize the cellular source, analyze expression of mRNA, and quantify protein synthesis of CCL24., Methods: Specimens of nasal inferior turbinates from controls and polypous tissue from patients suffering from chronic polypous sinusitis were collected. Furthermore, fibroblasts and epithelial cells were cultured. CCL24 protein was analyzed by immunohistochemistry and ELISA, expression of mRNA by SQ-RT-PCR., Results: CCL24 was observed in endothelial and epithelial cells. Specimens from patients expressed significantly (>2fold) more CCL24 mRNA than controls. Fibroblasts and unstimulated cells did not express CCL24 mRNA. Upon stimulation with TNF-alpha, INF-gamma, IL-4, or costimulation with TNF-alpha and INF-gamma CCL24 mRNA was significantly enhanced (3.2-19.6%). In controls, fibroblast, and unstimulated cells CCL24 protein was below detection limit. Most polyps comprised significant amounts of CCL24 (mean 0.24 ng/mg). TNF-alpha, INF-gamma or IL-4 induced CCL24 protein (0.1-0.3 ng/ml) in epithelial cells. Costimulation with TNF-alpha and IL-4 (0.1-30 and 1-30 ng/ml, respectively) synergistically induced synthesis of CCL24 protein (0.18-0.31 ng/ml)., Conclusion: In nasal polyps endothelial and epithelial cells are obviously the main source of CCL24, which was shown for transcription (mRNA) and production (protein) levels and was associated with diseases. Results gave evidence of CLL24- directed migration of cells from inside (the bloodstream) to the epithelial side (mucosa) in eosinophilic inflammatory diseases, e.g. nasal polyposis.

Background: Characteristic infiltrates of eosinophils are a hallmark of nasal polyps (NPs). Several studies suggest that members of the CC chemokine family may be involved in this process. RANTES (regulated on activation, normal t-cell-expressed and secreted) is a member of the CC chemokine family with chemotactic activity on mainly eosinophils and T lymphocytes. Thus, RANTES is an interesting target for the recruitment of eosinophils and T lymphocytes into the nose. The degree of the tissue eosinophilia has been reported to correlate with the severeness of the symptomatology of the disease and the extension on the lower respiratory tract, as well as with the probability of the recurrence of NPs. Therefore, we hypothesized that high numbers of eosinophils correlate with high levels of RANTES and that associated atopic diseases modify this correlation., Methods: Total RNA was extracted from NP homogenates, reverse transcribed and RANTES mRNA expression analyzed using semiquantitative reverse transcription polymerase chain reaction and Northern blot analysis. Histological studies divided NPs in an eosinophilic and low eosinophilic group. Additionally, RANTES protein concentration was measured in homogenates by a RANTES-specific enzyme-linked immunosorbent assay., Results: This study has clearly shown that RANTES is expressed and secreted in NPs. The group with a high tissue eosinophilia had a significant higher RANTES gene expression and protein production than NPs without tissue eosinophilia. The isolated coincidence of acetylsalicyl acid intolerance with chronic hyperplastic sinusitis/NP additionally increased significantly the RANTES amounts in NPs., Conclusion: Increased RANTES leads to increased tissue eosinophilia. Associated acetylsalicylic acid intolerance seems to enhance the amount of RANTES in NPs and might explain in part the more severe clinical course in those patients. Thus, RANTES appears to play an important role in mobilization of eosinophils into the local inflamed tissue.

Background: Polyposis, asthma, aspirin-intolerance and aspirin-triad are mostly accompanied with eosinophilia of mucosal airways. Chemotactic cytokines, the CC-chemokines regulated on activation, normal T-cell expressed (RANTES), eotaxin, and eotaxin-2 activate and attract eosinophilic leukocytes to the site of inflammation. This points to the implication of CC-chemokines in eosinophilia of nasal tissue of these diseases., Methods: Therefore, nasal polypous tissue specimens of patients suffering from chronic nasal polypous sinusitis (NP), intrinsic asthma (ATA), aspirin-intolerance (AINA), and aspirin-triad (TRIAD) were investigated. The amount of mRNA and protein of CC-chemokines was analyzed using semi-quantitative reverse transcriptase polymerase chain reaction and chemokine-specific enzyme-immuno-assays. The patterns of CC-chemokines were compared., Results: The mRNA-expression as well as protein synthesis of CC-chemokines was quantified in all tissues investigated. The expression of RANTES-mRNA in NP, ATA, AINA, and TRIAD (averaging 148-324% D-glyceraldehyde-3-phosphate dehydrogenase) and protein synthesis (0.13-0.15 ng/mg tissue weight) did not differ significantly. But the protein synthesis of eotaxin- and eotaxin-2-mRNA was significantly (P < 0.05) higher in TRIAD (3.3 pg/mg and 3.4 ng/mg tissue weight) (4 ng/mg tissue weight), than in NP, ATA, or AINA (1.8 pg/mg and 2.1 ng/mg, 2.1 pg/mg and 1.6 ng/mg, or 1.7 pg/mg and 2.2 ng/mg tissue weight, respectively)., Conclusion: Patients suffering from TRIAD in association with tissue eosinophilia were characterized by elevated eotaxin and eotaxin-2 mRNA-expression as well as protein-synthesis. This pointed to the implication of eotaxins and RANTES in eosinophilia-associated diseases. Further studies will have to prove, whether the analysis of these chemokines might improve the diagnosis of eosinophilia associated polyposis and initiate the development of new therapeutic strategies.

Objective: Macroscopically, tissue hydration in nasal polyps seems to be a phenomenon of unknown etiology. As the macromolecular composition of extracellular matrix (ECM) determines to a large extent tissue hydration, which in turn determines tissue volume, we investigated ECM components in nasal polyps (NP) in comparison to nasal mucosa of the inferior turbinate (TM) and sinus mucosa from patients with chronic sinusitis without polyps (CS)., Material and Methods: The following parameters were determined: (i) the dry weight of freeze-dried NP, TM and CS; (ii) the total protein content (Bio-Rad Protein Assay) of the tissue; (iii) the quality of proteins, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); (iv) the amount of albumin, using nephelometry; and (v) the amount of hyaluronic acid, using a chemical method: deaminative hydrolysis with carbazole., Results: In 20 NP we found a significantly elevated total protein content compared to TM (n = 20) and CS (n = 15), referred to 0.1 g dry of tissue. In SDS-PAGE of NP (n = 20) a protein band at approximately 70 kDa, representing albumin, dominated, in comparison to TM. The amount of albumin was significantly increased in NP compared to CS and TM. In contrast, the amount of the glycosaminoglycan, hyaluronic acid, was not elevated in NP or CS., Conclusion: Albumin was significantly increased in NP and CS, possibly as a result of inflammatory plasma exudation mechanisms. Hyaluronic acid seems to play no role in the tissue hydration of NP.

Human papillomavirus (HPV) infections are related to the genesis of various benign and malignant human neoplasias. The HPV types 16 and 18 seem to be causally related to the development of most squamous cell carcinoma of the anogenital tract and a proportion of carcinomas of the upper aerodigestive tract. The near 100% positivity of the HPV types 6 and 11 in laryngeal papillomatosis is well established. We investigated whether HPV also plays a role in non-neoplastic mucosal entities such as sinunasal polyposis, the genesis of which has been discussed as being triggered by viral infections. On DNA from 39 sinunasal polyps (33 patients), polymerase chain reaction (PCR) was performed using beta-globin primers for demonstration of amplifiable DNA in the tissue extracts. Consensus primers for the detection of several different HPV types were applied to the beta-globin-positive samples. The results were confirmed by Southern blot hybridization using consensus probes. Cycle sequencing was performed on the positive cases. All 39 samples showed positive signals for beta-globin. HPV-DNA investigations showed a slight positive signal in only 1 of the 39 investigated cases (2.6%). Further molecular investigations of this sample, including cycle sequencing, could not confirm this result. All the other tissue samples remained HPV-DNA-negative. Therefore, those HPV types readily detectable with the PCR primers and probes used are not frequently associated with sinunasal polyposis. The data confirm the hypothesis that HPV is correlated to a lesser extent to infectious mucosal lesions than to proliferative lesions. Furthermore, the results emphasize that the presence of HPV in specific lesions does not occur by chance, but represents a specific infection of the mucosa leading to proliferation and even to malignancy.

Background: Chronic sinusitis is one of the frequent inflammatory diseases and has a complex pathogenesis. A substantial factor seems to be recurrent bacterial infections. Pseudomonas aeruginosa (PA) frequently can be found in nasal smears of patients with persistent sinus symptoms after sinus surgery. Lately a new antimicrobial peptide of epithelial origin, human Beta-Defensin-2 (hBD-2), with a strong antibacterial effect against PA could be identified within lesional skin scales of patients suffering psoriasis. Aim of this study was to investigate hBD-2-mRNA expression in nasal cells and tissue., Methods: Total RNA was extracted from nasal polyps and turbinates following TRIzol protocol. Epithelial cells and fibroblasts of nasal human tissue were isolated and cultivated. The cells were stimulated with PA using different time points and different concentrations. Total RNA was isolated as mentioned above, reverse transcribed and amplified in a Semi-quantitative Reverse Transcriptase PCR (SQRT-PCR) with genespecific hBD-2 primers., Results: PA induces time- and dose-dependently hBD-2 gene expression in nasal epithelial cells. Unstimulated epithelial nasal cells were able to express hBD-2 mRNA constitutively, whereas nasal fibroblasts showed no hBD-2 mRNA expression. Nasal polyps showed a comparable less hBD-2 gene expression then nasal turbinates., Conclusions: hBD-2 possibly mediates a specific, early starting antimicrobial defense strategy of the nasal mucosa. This hypothesis would explain persistent infections with PA through diminished hBD-2 gene expression.

Eosinophilic tissue infiltration of nasal mucosa typical for allergic rhinitis and chronic polypous sinusitis may be due to chemotactic activity of chemokines specific for eosinophils. The CC-chemokines eotaxin, RANTES and MCP-3 have been postulated to be involved in the recruitment of eosinophils to certain inflamed tissues. To explore their possible role in chronic polypous sinusitis we examined eotaxin-, RANTES- and MCP-3-gene expression in human nasal polyps and normal human nasal mucosa of patients undergoing endonasal surgery for treatment of chronic polypous sinusitis. Using gene-specific primers in semi-quantitative reverse-transcriptase polymerase-chain-reaction experiments we found elevated expression of eotaxin- and RANTES-mRNA but no MCP-3-mRNA in non-atopic and atopic nasal polyps when compared to normal nasal mucosa.

Chronic polypoid sinusitis is often accompanied with bronchitis, asthma and various allergies. All of these disorders have immunologic defects in common. The present study was devised to measure the genetically determined parameter of alpha-1-protease inhibitor by isoelectric focusing. Genetic polymorphism of alpha-1-antitrypsin is the source for biochemically less active types of the protease inhibitor. In this investigation the alpha-1-protease inhibitor type of 86 patients from Niedersaxony and 95 patients from Schleswig-Holstein was determined. All patients required sinus surgery as treatment for chronic polypoid sinusitis. The alpha-1-protease inhibitor deficiency types PI-MS and PI-MZ were found five times more frequently in patients from Niedersachsony and three times more often in patients from Schleswig-Holstein than in a control (normal) group of 127 subjects (p < 0.01). These findings were comparable to data in other publications. Results clearly showed a connection between alpha-1-protease inhibitor deficiency types and chronic polypoid sinusitis. These findings indicate that a genetic predisposition to chronic polypoid sinusitis is somehow correlated with the gene locus for alpha-1-protease inhibitor.

Chronic sinusitis is a complex disease with a substantial health care impact. Multiple factors play a role in the pathogenesis of this disease, but the initial process is still not well understood. Characteristic infiltrates of eosinophils are a hallmark of this disease. The selective appearance of T lymphocytes and eosinophils in growing polyps seems to be mediated by locally produced CC chemokines in cooperation with proinflammatory cytokines. Both are able to induce vascular adhesion molecules and attract rolling eosinophils as haptotaxins to the site of inflammation. Associated atopic diseases, like asthma or perennial allergies, have a higher eochemoattractant activity, which may contribute either to the nasal or bronchial mucosal accumulation of eosinophils. This could potentiate the eosinophilic effects in both tissues and possibly explain the severer symptomatology and higher recurrence rate of nasal polyps. RANTES, eotaxins 1 and 2 and IL-5 certainly play a role in chronic sinusitis. To our mind, the exact part of each mediator does not seem to be important, because just the complex cooperation of chemokines and also cytokines allows the underlying inflammatory process to become a chronic sinusitis. Therapeutic targets for eosinophil-related diseases are the impairment of CC chemokine expression and release by glucocorticoids, proteolytic degradation by endogenous enzymes, inhibition/antagonism of the CCR-3 and also endogenous antagonism by other chemokines. [ABSTRACT FROM AUTHOR]