Etiology of nasal polyps: an update

Nasal polyps are the results of chronic inflammation of nasal mucosa. Ethiopathology of nasal polyposis is still unclear and investigated. The main aim of this study was the assessment of interleukin 2 (IL-2), 4 (IL-4) and 13 (IL-13) and interferon gamma (IFN-gamma) expressions in two groups of patients with nasal polyps: non-allergic and allergic to dust. Twelve patients with nasal polyposis were selected, six of them allergic and six non-allergic. Patients with allergy were distinguished from those without allergy on the basis of positive allergy skin tests to dust and serum levels of IgE. Blood sample was obtained from patients and examined for the expression of Interleukin 2, 4, and 13 and IFN-gamma by intracellular staining procedure after stimulation with PMA/ionomycin (phorbol 12-myristate 13-acetate) and allergen (D. pteronyssimus, Allergopharma). Negative correlation was found between expression of Interleukin 2, 4, 13 after PMA/ionomycin and allergen stimulation (p > 0.05) into two groups. Although mean expression of IFN-gamma and IL-2 were lower in allergic patients with nasal polyps. Mean value of IL-4 and IL-13 were higher in allergic patients in comparison to non-allergic, but the correlations were not significant. We did not find correlations between expression of investigated interleukins and bronchial asthma and ASA-intolerance existed in patients (p > 0.05). We found that expression of IFN-gamma was significantly lower in patients with polyps recurrences followed by polypectiomies (p = 0.05). In patients who used intranasal local steroid before treatment we observed significant decrease of 11-2 and 11-13 levels after allergen stimulation (p = 0.,034, p = 0.023). No specific differences of expression of investigated cytokines between allergic versus non-allergic patients suggests that the allergic mechanism may not play an fundamental role in the aetiology and formation of nasal polyps. Furthermore local steroids seem to be used in every patient after polypectomy because of their anti-inflammatory stimulation.     

白介素-5在变应性鼻炎鼻黏膜中的表达及意义

目的：了解白介素 5（IL 5）在变应性鼻炎鼻粘膜内浸润的炎性细胞中的表达及分布,探讨IL 5与嗜酸性粒细胞积聚及发病机制的关系。方法：采用免疫组化染色法（SP法）对变应性鼻炎下鼻甲粘膜（A组）、单发鼻息肉组织（B组）、无变应性鼻炎下鼻甲粘膜组织（C组）切片进行IL 5染色,对IL 5染色阳性细胞计数分类,对统计结果行方差分析。结果：A、B两组组织中可见较多的IL 5阳性染色细胞,多见嗜酸性细胞、淋巴细胞、中性粒细胞等阳性染色,两组IL 5阳性细胞总数、Eos计数无统计学差异,但均高于C组（P＜0.01）。结论：IL 5在变应性鼻炎鼻粘膜、鼻息肉组织内浸润的多种炎性细胞中表达,能客观反映免疫或炎症反应的程度,可作为变应性鼻炎诊断评分系统的补充。     

VCAM-1表达在上下呼吸道一致性的研究

目的：探讨血管内皮细胞黏附分子（VCAM-1）在呼吸道变应性炎症患者中的表达和意义。方法：将39名患有变应性鼻炎并哮喘的患者和21例仅患有变应性鼻炎的患者分成两组,取鼻腔黏膜和支气管黏膜进行活组织病理检查,并用免疫组织化学的方法对VCAM-1的表达进行检测。结果：变应性鼻炎并哮喘组支气管黏膜中嗜酸粒细胞数显著高于变应性鼻炎组（t=12．81,P〈0．01）,变应性鼻炎组鼻腔黏膜中VCAM-1阳性细胞比例与变应性鼻炎并哮喘组相比差异无统计学意义。变应性鼻炎并哮喘组支气管黏膜中VCAM-1阳性细胞比例显著高于变应性鼻炎组（t=9．43,P〈0．01）,变应性鼻炎并哮喘组支气管黏膜中VCAM-1阳性细胞比例和支气管黏膜嗜酸粒细胞数呈显著正相关（r=0．788,P〈0．01）。结论：VCAM-1表达的上调,导致呼吸道变应性炎症的蔓延。     

转化生长因子β1白细胞介素-6及11和17在变应性鼻炎患者鼻黏膜中的表达

目的：探讨转化生长因子β1（TGF-β1），白细胞介素（IL）-6、11和17在变应性鼻炎患者鼻黏膜中的表达及意义。方法：采用酶联免疫吸附试验定量检测19例变应性鼻炎患者（变应性鼻炎组）下鼻甲黏膜和21例行鼻中隔偏曲矫正术的非变应性鼻炎患者（对照组）下鼻甲黏膜组织中TGF-β1，IL-6、11和17的蛋白表达量。结果：①变应性鼻炎组的TGF-β1蛋白表达量显著高于对照组（P〈0．01）；②IL-6、11和17的蛋白表达量在两组间差异无统计学意义（均P〉0．05）；③TGF-β1，IL-6、11和17的蛋白表达量在并发哮喘和不并发哮喘的变应性鼻炎患者间的表达差异无统计学意义（均P〉0．05）。结论：TGF-β1可能在变应性鼻炎的组织结构重塑中起作用，而IL-6、11和17可能在此过程中无明显作用，这可能是变应性鼻炎和哮喘组织结构重塑差异的分子基础之一。     

Immunoelectron microscopic findings in patients with allergic rhinitis

BACKGROUND: In middle Europe the prevalence of allergic rhinitis is up to 15 % to 25 %. Allergic rhinitis is characterised by an inflammation of the nasal mucosa induced by different allergens. The patients suffer from symptoms like sneezing, rhinorrhea and nasal airway obstruction caused by morphological changes of the nasal mucosa. This symptomatology is considered to be a result of accumulation and activation of inflammatory cells. Further some neuropeptides like Calcitonin Gene Related Peptide (CGRP) and Substance P (SP) play an additional role in pathophysiology of allergic rhinitis. PATIENTS AND METHODS: Tissue samples from 28 human turbinates of patients with perennial rhinitis were taken during nasal surgery and preserved in phosphate-buffered glutaraldehyde or paraformaldehyde. Ultrathin sections were cut. The samples were dehydrated and embedded in Araldit. After polymerization an immunocytochemical staining-technique using a gold-labeled antibody was carried out. Immunostained structures were photodocumented by using a transmission electron microscope. RESULTS: In the lamina propria mucosae an extensive edema and several inflammatory cells like lymphocytes, plasma cells, eosinophiles and macrophages was found. The capillaries showed an activated endothelium. Immunoreactive nerve fibers were found in the periglandular tissue around the acini, ducts and in the glandular connective tissue. Neuroglandular synapses with dense core vesicles and positive immunoreactions to CGRP and SP could be detected. Neuropeptidergic axons were often observed near to plasma cells. CONCLUSIONS: In the edematous nasal mucosa an infiltration with different inflammatory cells was found. Using electron microscopical techniques nerve structures near the submucosal glands could be demonstrated. Immunoreactions to the neuropeptides CGRP and SP were detected in the periglandular nerves and in neuroglandular synapses. These findings demonstrate the direct nerve control of glandular functions in allergic rhinitis. CGRP is generally known to have a vasodilatatory effect and to stimulate the secretion of nasal seromucous glands. In addition, SP as a short-acting vasodilatator may induce vascular permeability and glandular secretion. These immunoelectron microscopical findings further elucidate pathomorphological mechanisms in allergic rhinitis.     

Heme oxygenase (HO)-1 is upregulated in the nasal mucosa with allergic rhinitis

BACKGROUND: Heme oxygenase (HO) is considered to be an antioxidant enzyme that catabolizes heme to produce carbon monoxide (CO) and biliverdin. Three isoforms of HO have been discovered. Recently, HO-1 has been found to be upregulated after allergic inflammations of the lower airway. OBJECTIVE: The objective of this study was to address the expression of HO isoenzymes 1 and 2 in the nasal mucosa of patients with allergic rhinitis as well as normal control subjects. METHODS: Nasal mucosa from 30 patients with persistent allergic rhinitis as well as from 10 normal volunteers was used in this study. We used immunofluorescent technique, Western blotting, and real-time quantitative polymerase chain reaction to localize and quantify the expression of these isoenzymes in normal and allergic human nasal tissues. RESULTS: We found that HO-1 is expressed in the epithelial cells of seromucinous glands and macrophages with significant upregulation of its glandular expression in allergic rhinitis but with no difference in its macrophage expression between the study groups in contrast to HO-2 that is expressed in the vascular endothelial lining cells as well as macrophages with no marked difference between the study groups. CONCLUSION: We demonstrated that expression of HO-1, but not HO-2, was upregulated within the nasal tissues in allergic rhinitis inflammation, and understanding the induction of HO-1 expression may provide for better management of allergic rhinitis that involves oxidative stress.     

白细胞介素12基因治疗小鼠变应性鼻炎的实验研究

目的探讨鼻腔局部应用EB病毒(Epstein-Barrvirus,EBV)质粒载体介导的白细胞介素12(interleukin-12,IL-12)基因治疗对变应性鼻炎炎症反应的调节作用。方法将36只6～8周雄BALB/C实验小鼠随机分为变应性鼻炎组、IL-12基因治疗组和正常对照组,每组12只。以BALB/c小鼠经卵清蛋白(ovalbumin,OVA)免疫建立变应性鼻炎模型,用阳离子脂质体包裹EBV质粒载体介导的IL-12表达质粒(pGEG.mI-L12)形成混合物EBV/lipoplex,于激发前鼻腔局部滴入后,观察小鼠变应性症状的改善情况,并检测该基因在3组实验鼠鼻黏膜局部的表达情况以及对鼻黏膜炎性细胞和Th2细胞因子的影响。结果基因治疗组小鼠鼻黏膜中IL-12mRNA阳性细胞数量明显高于变应性鼻炎组,差异有统计学意义(P<0.01);基因治疗组小鼠鼻黏膜中IL12阳性细胞数量明显高于变应性鼻炎组(P<0.05);变应性鼻炎组鼻黏膜中嗜酸粒细胞、肥大细胞和IL5阳性细胞比例显著高于基因治疗组和正常对照组(P<0.01);变应性鼻炎组外周血中总IgE含量显著高于正常对照组和基因治疗组(F=1216.21,P<0.01)。结论鼻腔局部应用EBV/lipoplex后,pGEG.mIL-12能够在鼻黏膜中高效地表达,能明显抑制鼻腔的变应性反应。EBV/lipoplex有望成为变应性鼻炎免疫治疗一种新的方法。     

The concentration and expression of IL-5 in human nasal polyp tissue]

OBJECTIVE: To study the concentration and expression of IL-5 in nasal polyp tissues and explore its significance in the micro-environment differentiation of eosinophils accumulation and clarify the conception of nasal polyposis. METHODS: The 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 was used as control. RESULTS: 1. IL-5 concentration in the polyp tissues was significantly higher than that in inferion turbinate mucosa(P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentration in polyp tissues was markedly higher in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). IL-5 concentration had no correlation with age and sex (P > 0.05). 2. 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of the lymphocytes and neutrophils were IL-5 positive, and 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 inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissues was significantly stronger in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and inferior turbinate nasal mucosa (both P > 0.05). CONCLUSION: IL-5 is a key protein in eosinophilic pathologic mechanisms in nasal polyp tissues.     

黏蛋白在鼻息肉及变应性鼻炎中的基因表达水平的研究

目的：探讨5种人黏蛋白基因（MUC2、MUC5AC、MUC5B、MUC18、MUC19）在鼻息肉和变应性鼻炎中的基因表达水平及临床意义。方法：采用RT-PCR及实时荧光定量RT-PCR,检测35例鼻息肉、18例变应性鼻炎患者下鼻甲及18例正常人黏蛋白基因（MUCS）在下鼻甲黏膜上皮的基因表达水平。结果：RT-PCR及实时荧光定量RT-PCR的结果表明,MUC5AC、MUC5B在鼻息肉组和变应性鼻炎组中的表达高于对照组（P〈0.05）,鼻息肉组与变应性鼻炎组及对照组间表达差异均无统计学意义（P〉0.05）。MUC2、MUC18在3组中的表达差异无统计学意义（P〉0.05）。MUC19在变应性鼻炎组中的表达高于鼻息肉组和对照组（P〈0.05或P〈0.01）。结论：MUC5AC和MUC5B在鼻息肉、变应性鼻炎的黏液过度分泌过程中具有重要意义;MUC19在变应性鼻炎的黏液过度分泌过程中也有重要意义,但是对鼻息肉患者的黏液分泌没有明显作用;未发现MUC2、MUC18在变应性鼻炎的黏液过度分泌过程中起明显作用。     

吸入性氢气清除法鼻粘膜微循环血流量测定

测定国人鼻粘膜微循环血流量。方法 应用Ａｍｅｆｌｗ－Ｈ２微循环测量仪对１３０例鼻粘膜微循环血流一在局部喷雾血管收缩剂前后进行了测量，其中健康成人９８例，鼻息肉患者１８例，应变性鼻炎患者１４例。统计学处理采用ｔ检验。结果 健康成人对照组鼻粘膜微循环血流量大于鼻息肉组及变应性鼻炎组。     

Studies on the function of mast cells infiltrating in nasal polyps]

The function of nasal polyp mast cells has not been elucidated despite the large number of these cells observed in tissues. We examined these mast cells histochemically, immunohistologically and functionally. Ninety-three percent of collagenase dispersed cells in a nasal polyp were formalin-sensitive. These dispersed cells released histamine in reaction to calcium ionophore A23187 in a dose dependent manner, but not in response to C5a, Compound 48/80 or Substance P. From these results, dispersed mast cells from nasal polyps were considered to be analogous to dispersed mast cells from the human lung and nasal mucosa but not those from human skin. On the other hand, in the reaction with anti-human IgE, dispersed mast cells from a non allergic nasal polyp could not be seen to release histamine. In only 2 of 7 patients, could histamine release in response to Japanese red cedar antigen, from mast cells sensitized passively with the serum of Japanese red cedar pollinosis, seen. Using small tissue samples from polyps, histamine was released by anti-human IgE in allergic patients but not in non allergic patients. Immunohistologically in allergic nasal polyps, some IgE positive mast cells could be seen, whereas in non allergic polyps these cells were absent. These observations suggest that mast cells which had accumulated in nasal polyps both with and without allergy were capable of functional histamine release, whereas in the nasal polyps of allergy patients but not in non-allergic patients these cells are involved in IgE mediated reactions.     

Distribution of rantes and interleukin-5 in allergic nasal mucosa and nasal polyps.

Eosinophil-chemoattracting cytokines are thought to be important in the pathogenesis of allergic inflammation. However, little is known about the presence and significance of RANTES in nasal allergy and nasal polyps, two well-known rhinologic disorders characterized by eosinophil infiltration in the tissue. In order to evaluate the role of RANTES in eosinophil infiltration in vivo, the tissue distributions of RANTES and interleukin-5 (IL-5) and their correlation with eosinophil infiltration were investigated. Nasal mucosa specimens were obtained from 9 allergic and 12 control subjects, and nasal polyps from 6 allergic and 9 nonallergic subjects. All the subjects were divided into 4 groups: normal mucosa, allergic mucosa, nonallergic polyps, and allergic polyps. To identify the cellular localizations of RANTES and IL-5, we used specific immunohistochemical staining. We also investigated the differences in cytokine expression among the 4 groups, and the correlation between cytokine expression and eosinophil infiltration in the tissue. RANTES was expressed in the epithelium, endothelium, and some submucosal cells, while IL-5 was confined to the cells in the submucosa. Expression of both RANTES and IL-5 significantly increased in allergic mucosa and nasal polyps compared to normal mucosa; however, there was no significant difference in their expression between allergic and nonallergic polyps. Both cytokines had a significant correlation between their expression and either total or activated eosinophil numbers. The results of this study suggest that RANTES, as well as IL-5, plays a role in eosinophil recruitment in allergic nasal mucosa and nasal polyps in vivo.     

Expression and localization of TRPV1 in human nasal mucosa

Capsaicin is the pungent principle in chili peppers and previous studies reported that topical application of capsaicin to patients with allergic and non-allergic rhinitis produced significant and long-lasting relief of symptoms. The capsaicin receptor (TRPV1, VR1) is a nociceptive transducer and the existence of TRPV1 in non-neuronal cells as well as neuronal cells has been reported. In order to clarify the role of TRPV1 on the upper airway, we examined the localization and the expression of TRPV1 in human nasal mucosa. Surgically obtained human nasal specimens were processed for immunohistochemistry with commercial anti-TRPV1 antibody. We also performed immunofluorescence with anti-TRPV1 antibody and anti-neurofilament antibody or anti-CD31 antibody. Epithelial cells and vascular endothelial cells were cultured from nasal turbinates, respectively. For RT-PCR analysis, total RNA was isolated, and then RT-PCR was performed. Immunohistochemical studies revealed that TRPV1 positive cells were found on epithelial cells, vascular endothelial cells, submucosal glands and nerves in human nasal mucosa. By RT-PCR analysis, the mRNA expression of TRPV1 was confirmed in human nasal mucosa. These results suggest that capsaicin can directly influence the epithelial secretory and various functions via TRPV1 as well as the activation of the sensory neurons.     

Isolation and phenotypic characterization of mucosal nasal lymphocytes by direct ex vivo analysis

Cellular inflammation of the nasal mucosa demonstrates a local immune response which plays an important role in allergic rhinitis. The aim of the present study was to characterize nasal mucosal lymphocytes regarding their activation and differentiation state by direct ex vivo flowcytometric analysis. Lymphocytes from the inferior turbinates were isolated by a mechanical method of preparation and, for comparison, from peripheral blood by Ficoll gradient centrifugation. Patients suffering from rhinitis or difficulty in nasal breathing were divided into an allergic (pollen-allergy, n = 13) and non-allergic group (n = 24). Expression of different T- and B-cell markers was determined by flowcytometric analysis. CD4+ T-cells from the nasal mucosa exhibited a memory phenotype (CD45RO+, 97%), were highly activated (CD69+, 43–73%), and showed low expression of the cutaneous lymphocyte antigen (CLA+, 5%). Nasal CD20+ B-lymphocytes expressed significantly higher levels of mIgE and lower levels of CD23 and CD80 than peripheral B-cells. Subsets of CD80+ (4%) and CD86+ (6%) CD20+ B-lymphocytes were identified in the nasal mucosa. No significant differences between allergic and non-allergic individuals were determined. As expected, the data show profound phenotypical differences between circulating peripheral blood and nasal mucosal lymphocytes. Activated memory lymphocytes are present in the nasal mucosa from allergic, but also non-allergic patients and may indicate to a significant role of a local inflammatory state without systemic criteria for allergy. In our study, we show that direct ex vivo isolation of lymphocytes is practicable method and offers a new technique to examine the local nasal allergic immune response using a multiparametric phenotypical analysis. Christin Wolfram and Claudia Rasche contributed equally to this work.     

Nasal lavage cytology and mucosal histopathological alterations in patients with rhinitis

IntroductionThe extent of epithelial lesion in allergic and non-allergic rhinitis and its association with inflammatory changes in nasal lavage has not been clarified.ObjectiveTo verify the association between the inflammatory cells in the nasal lavage, epithelial lesion extent and basement membrane thickness, in the nasal mucosa of patients with rhinitis; to determine the cutoff point of the percentage of eosinophils in the nasal lavage associated with the atopic patients.MethodsPatients with rhinitis and indication for septoplasty and (or) turbinectomy for turbinate hypertrophy were selected, and were submitted to allergy skin tests, nasal lavage with measurement of albumin and interleukin-8 levels, total and differential counting of cells, and mucosal histopathological analysis to determine the extent of epithelial lesion, and degree of basement membrane thickening.ResultsFifty-six patients with a median age of 24.5 years and a diagnosis of allergic rhinitis (n = 36) and non-allergic rhinitis (n = 20) were studied. In atopic subjects, allergy skin tests were positive for Dermatophagoides pteronyssinus in 35 (97.0%) and Lolium perenne in 18 (50.0%). Atopic subjects showed a higher clinical score index of rhinitis compared to non-atopic ones. The total count of cells, neutrophils, and levels of albumin and IL-8 were not different in the nasal lavage of atopic and non-atopic subjects. The cutoff point for eosinophil count in nasal fluid for the distinction between allergic rhinitis and non-allergic rhinitis was 4%. Some degree of epithelial lesion was more frequent in allergic rhinitis (94%) than in non-allergic rhinitis (65%) patients. In the presence of basement membrane thickness, as a marker of remodeling, there was no difference in the nasal lavage of patients with allergic rhinitis and non-allergic rhinitis.ConclusionIn this series, 4% was the cutoff point for the number of eosinophils in the nasal lavage, for atopy differentiation. Upper airway remodeling accessed by basement membrane thickness showed similar inflammatory cell infiltrate in the nasal lavage, regardless of the presence of atopy.     

Expression of annexin A1 in normal and chronically inflamed nasal mucosa

OBJECTIVE: To examine the expression pattern of annexin A1 in normal and chronically inflamed nasal mucosa to investigate its possible role in nasal inflammation. DESIGN: Immunohistochemical analysis. SUBJECTS: Samples of middle turbinates from 5 healthy subjects and 5 patients with perennial rhinitis, and samples of nasal polyps from 7 patients. INTERVENTIONS: Annexin A1 expression was examined with a standard immunohistochemical protocol on paraffin-embedded sections. RESULTS: Annexin A1 was highly expressed by ciliated cells, where it was concentrated on the apical surface and within the cilia. Goblet cells and nondifferentiated basal epithelial cells did not stain. In the glands of the lamina propria, intense staining was found in the cytoplasm and in the nuclei of the cells in the duct epithelium, whereas acinar cells did not stain. Intense cytoplasmic staining was observed in infiltrating polymorphonuclear cells and macrophages. No differences in the pattern or the level of expression of annexin A1 were found in the epithelial cells and glands of normal and chronically inflamed (perennial rhinitis or polyps) nasal mucosa. CONCLUSION: These results suggest that the expression of annexin A1 in respiratory epithelium of nasal mucosa is related to cell type and differentiation status of the cells and is not significantly altered by inflammatory diseases.