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.     

鼻息肉中单核细胞趋化蛋白1和血管内皮生长因子的表达

目的明确单核细胞趋化蛋白1（monocyte chemotactic protein 1，MCP-1）和血管内皮生长因子（vascular endothelial growth factor，VEGF）在鼻息肉组织中的表达及其相关性，初步探讨MCP-1与鼻息肉发生的关系。方法取40例鼻息肉组织和25例下鼻甲组织，应用原位杂交和免疫组织化学等方法检测MCP-1和VEGF mRNA及蛋白质的表达。结果鼻息肉组织中MCP-1和VEGF mRNA及蛋白质的表达均高于对照组下鼻甲组织（P值均〈0．01）；鼻息肉组织中MCP-1和VEGF蛋白质的表达呈正相关（r=0．871，P〈0．05）。结论鼻息肉组织中MCP-1和VEGF表达增加，二者协同作用可能是鼻息肉形成的原因之一。     

鼻息肉中免疫球蛋白E白细胞介素5和粒细胞巨噬细胞集落刺激因子的表达

目的 :探讨鼻息肉组织中免疫球蛋白E(IgE)、白细胞介素 5 (IL 5 )和粒细胞巨噬细胞集落刺激因子(GM CSF)的表达对嗜酸性粒细胞 (EOS)浸润聚集的作用及IgE与IL 5和GM CSF的关系及其意义。 方法 :应用连续两步免疫酶法 (Sandwich)检测 31例鼻息肉标本 (鼻息肉组 )和 11例下鼻甲黏膜标本 (对照组 )组织匀浆中IgE水平 ;应用酶联免疫吸附测定法检测两组组织匀浆中IL 5和GM CSF水平 ,同时观测两组组织中EOS的浸润程度。结果 :鼻息肉组IgE、IL 5和GM CSF水平均明显高于对照组 (圴 P <0 .0 1) ,且三者水平分别与鼻息肉组织中EOS浸润数呈正相关 (r分别为 0 .73、0 .5 4和 0 .4 8,均P <0 .0 1) ,且鼻息肉中IgE与IL 5和GM CSF水平呈正相关 (r分别为 0 .6 5和 0 .4 2 ,分别P <0 .0 1和P <0 .0 5 )。结论 :IgE局部水平的上调提示鼻息肉中存在局部变态反应 ,其对鼻息肉中IL 5和GM CSF的表达上调和EOS的聚集具有重要作用 ;IL 5和GM CSF在鼻息肉中高表达 ,对EOS的浸润聚集具有重要作用。     

鼻息肉组织中固生蛋白C的表达与转化生长因子β1的关系

目的探讨细胞外基质(extracellularmatrix,ECM)成分固生蛋白C(tenascinC,TNC)在鼻息肉组织中异常表达的原因。方法采用免疫组化方法检测TNC和转化生长因子β1(transforminggrowthfactor-β1,TGF-β1)在鼻息肉组织中的表达和关系。进一步采用细胞培养、实时定量RTPCR和原位ELISA技术研究TGFβ1及嗜酸粒细胞对人呼吸道上皮细胞系BEAS2B细胞TNC表达的调控作用。结果①TNC和TGF-β1蛋白在鼻息肉组织中的表达显著上调,TNC表达强度与TGFβ1阳性细胞总数(r=-0.58,P<0.01)和TGFβ1阳性的嗜酸粒细胞数显著相关(r=-0.61,P<0.01);②浓度为1ng/ml和10ng/ml的TGFβ1刺激4h后BEAS2B细胞TNCmRNA的表达分别为未刺激状态下的(7.20±3.43,x±s,下同)倍和(22.48±5.35)倍,与未刺激状态下的表达水平相比差异有统计学意义(P值<0.01);与刺激24h后TNC蛋白表达的荧光强度相比差异亦有统计学意义(P<0.05);③BEAS2B细胞和嗜酸粒细胞以2∶1、1∶1和1∶2的数量比例进行共培养,4h后BEAS2B细胞TNCmRNA的表达与未刺激状态下的表达水平相比,差异均有统计学意义(P值均<0.01);24h后TNC蛋白表达的荧光强度与未刺激状态下的荧光强度相比差异亦均有统计学意义(P值均<0.05);嗜酸粒细胞的这种诱导作用可以被抗TGFβ1的中和抗体显著抑制(P<0.05)。结论TGF-β1和嗜酸粒细胞可以诱导呼吸道上皮细胞对TNC的表达,嗜酸粒细胞的作用部分通过TGFβ1介导,鼻息肉组织中TNC表达的增高同嗜酸粒细胞来源的TGF-β1有关。     

Immunological features of patients with polypous ethmoiditis

34 patients with nasal polyps (NP) and 12 normal individuals were studied immunologically to investigate association of nasal polyp formation with disorders of general and local immunity. In NP patients there were decreased preoperative and early postoperative levels of peripheral blood T- and B-lymphocytes, functional activity of lymphocytes and neutrophils. In contrast to normal individuals, nasal secretion of NP patients contained degenerative epithelial cells and neutrophils, activated lymphocytes, monocytes and eosinophils. Nasal polyp tissues obtained after polypectomy contained more B-lymphocytes than T-lymphocytes. Lymphocytes and neutrophils in nasal polyps had elevated functional activity in tissue culture. We conclude that local hyperactivation of T- and B-lymphocytes as well as neutrophils contribute much to nasal polyp formation.     

Eosinophil chemoattractants and related factors in nasal polyps

OBJECTIVE: In vitro studies and animal experiments have shown that cytokines and chemokines are closely related to eosinophil migration, activation, and survival. It remains controversial, however, whether some chemokines or cytokines are actually responsible for the accumulation of eosinophils in nasal polyp tissues. We studied cytokines and chemokines in nasal polyp tissues taken from patients with chronic rhinosinusitis to clarify the pathogenesis of eosinophil accumulation. MATERIALS AND METHODS: Nasal polyp tissues obtained from 20 patients with chronic rhinosinusitis were studied. Concentrations of interleukin (IL-) 5, IL-13, eotaxin, regulated upon activation in normal T cell expressed and secreted (RANTES), and thymus and activation-regulated chemokine (TARC) in homogenates of polyp tissues were measured by ELISA. Nasal polyp tissues were stained by hematoxillin and eosin and were immunostained by an antibody against EG2. The numbers of eosinophils and immunopositive cells for EG2 in the submucosal layer were counted using a microscope. RESULTS: No significant differences were seen in the numbers of eosinophils and EG2-positive cells, or in the concentration of IL-5, eotaxin, TARC, RANTES in nasal polyp tissues between patients with and without atopic predisposition. Significant positive correlations existed, however, between the number of eosinophils and IL-5, eotaxin, and TARC concentration. IL-13 concentration was below detection in all patients. CONCLUSION: We hound that IL-5, eotaxin, and TARC may play an important role in the accumulation of eosinophils in nasal polyps regardless of the presence of atopic predisposition.     

白细胞介素-5及嗜酸性粒细胞在鼻息肉中的相关性研究

目的探讨白细胞介素-5(IL-5)在鼻息肉中的表达及其在鼻息肉形成与发展中的作用。方法采用双抗体夹心酶联免疫吸附测定法检测31例鼻息肉患者、11例慢性鼻窦炎患者及6例正常人组织匀浆中IL-5含量;对15例鼻息肉、11例慢性鼻窦炎及6例正常人进行HE切片染色,以观察组织中嗜酸性粒细胞的数量。结果①鼻息肉、慢性鼻窦炎及正常对照组IL-5含量(pg／mg)分别为23．44±6．68,16．41±3．09,12．86±4．17,鼻息肉组织IL-5含量高于慢性鼻窦炎及正常对照组(P<0．01)。②鼻息肉、慢性鼻窦炎及正常对照组的苏木精伊红染色嗜酸性粒细胞数分别为7．42±2．33,1.30±0．59,1．07±0．70,鼻息肉组嗜酸性粒细胞数高于慢性鼻窦炎和正常对照组(P<0．05)。结论鼻息肉是以嗜酸性粒细胞浸润为特征的疾病过程,对嗜酸性粒细胞有活化作用的IL-5对鼻息肉的形成起着一定的作用。     

天然免疫蛋白PLUNC在鼻息肉中的表达及与鼻息肉临床特征的关联性

目的：探讨天然免疫蛋白腭、肺、鼻上皮克隆（PLUNC）在鼻息肉组织中的表达情况,分析PLUNC蛋白浓度与鼻息肉大小（鼻内镜记分）和手术后复发的关联性。方法：采用免疫组织化学和实时定量PCR检测鼻息肉组（28例鼻息肉患者,其中13例为术后复发患者）和对照组（16例钩突黏膜对照）PLUNC的组织染色定位和mRNA的表达情况,同时采用ELISA检测初发和复发鼻息肉组织中PLUNC的蛋白浓度差异,评估PLUNC蛋白浓度与息肉大小、鼻塞、流涕症状记分的相关性。结果：鼻息肉组织中PLUNC主要定位在黏膜上皮和腺体,染色强度记分显著低于对照的钩突组织（P〈0．01）;PLUNCmRNA表达水平也显著低于对照的钩突组织（P〈0．01）。初发和复发鼻息肉组织中PLUNC蛋白OD值分别为0．33±0．11和0．15±0．05,差异有统计学意义（P〈0．01）。小型和大型鼻息肉PLUNC蛋白OD值分别为0．32±0．14和0．19±0．07,差异有统计学意义（P〈0．05）。鼻息肉组织中PLUNC蛋白浓度与鼻塞和流涕记分存在显著负相关（r=-0．51和r=-0．57,P〈0．01）。结论：PLUNC的表达降低提示鼻息肉的发生可能与天然免疫反应的减弱有关,因此,上调天然免疫分子如PLUNC等的表达可作为干预鼻息肉发病的一个新策略。     

Inducible cyclooxygenase and interleukin 6 gene expressions in nasal polyp fibroblasts: possible implication in the pathogenesis of nasal polyposis

BACKGROUND: Inflammation is believed to be related to the pathogenesis of nasal polyp (NP). Inducible cyclooxygenase (COX-2) and interleukin (IL) 6 are important mediators of inflammation. However, no information is available regarding the expression of these mediators in nasal polyp fibroblasts (NPFs). The inductive effects of proinflammatory cytokines (IL-1alpha or tumor necrosis factor alpha) alone or in combination with prostaglandin E(2) on IL-6 and COX-2 messenger RNA (mRNA) synthesis in NPFs were investigated. DESIGN: The expressions of IL-6 and COX-2 mRNAs in NPFs and in 34 surgical specimens of NP were detected by Northern blot and in situ hybridization. RESULTS: Significant amounts of constitutive IL-6 and COX-2 mRNAs were produced in NPFs. Cytokines induced IL-6 and COX-2 mRNA synthesis in NPFs. Meloxicam (a specific COX-2 inhibitor) suppressed the induction of cytokines on IL-6 mRNA levels, and these effects could be reversed by exogenous prostaglandin E(2). In situ hybridization revealed that IL-6 and COX-2 mRNAs were detected primarily in fibroblasts, macrophages, and plasma cells. Aggregation of plasma cells as well as collagen deposition in vicinity to IL-6 mRNA-producing fibroblasts was found. Rich vascularity around COX-2 mRNA(+) fibroblasts was also identified. CONCLUSIONS: The pathogenesis of nasal polyposis involves NPFs through synthesizing IL-6 to modulate the activation of immune responses (plasma cell formation) and synthesis of stroma. Inducible cyclooxygenase also contributes to NP development by promoting vasodilatation and modulating the cytokine-induced IL-6 gene expression in NPFs.     

金黄色葡萄球菌肠毒素B对人鼻黏膜上皮细胞的促炎作用

目的探讨金黄色葡萄球菌肠毒素B（staphylococcus aureus enterotoxin B，SEB）对原代培养的人鼻黏膜上皮细胞释放促炎因子和（或）趋化因子的作用。方法将无血清原代培养的人鼻息肉及下鼻甲上皮细胞分别在SEB1、10、100ns／ml，白细胞介素（intedeukin，IL）1β 20ng／ml及SEB 10ng／ml＋地塞米松13ng／ml等不同条件下孵育，12、24和48h后采用原位杂交方法检测鼻黏膜上皮细胞分泌的IL-5和粒细胞巨噬细胞集落刺激因子（granuloyte macrophagc colony stimulating factor，GM-CSF）在mRNA水平的变化。结果①SEB刺激鼻黏膜上皮时，IL-5和GM—CSF mRNA表达增加，在一定范围内呈现时间和剂量依赖性，以10ng／ml、24h时最明显（P〈0．05），且鼻息肉组的表达高于下鼻甲组，差异具有统计学意义（P〈0．05）；②IL-1β 20ng／ml组IL-5和GM-CSF mRNA表达增加不及SEB 10ng／ml组明显，差异具有统计学意义（P值均〈0．05）；③SEB 10ng/ml＋地塞米松13ng／ml组的IL-5和GM-CSF mRNA表达强度较SEB 10ng／ml组弱，差异具有统计学意义（P值均〈0．05）。结论SEB对原代培养的人鼻黏膜上皮细胞具有促炎作用。     

Expression and localization of the inducible isoform of nitric oxide synthase in nasal polyps

Nitric oxide (NO) and peroxynitrite play an important role in pathophysiology of several airway diseases. An inducible isoform of nitric oxide synthase (iNOS) is known to be expressed in the nasal mucosa in allergic and chronic rhinitis. Few reports exist, however, on the expression of iNOS in nasal polyps. We detected and localized iNOS expression in nasal polyp tissue. Nasal polyps were obtained from 10 patients following polypectomy, and divided into allergic and infectious groups based on clinical presentation and laboratory testing. One nasal mucosa of the inferior turbinate was also obtained from a cadaver without nasal disease. iNOS expression was studied by immunohistochemistry under light and electron microscopy. Immunoreactivity for iNOS was localized to the mucosal epithelium, inflammatory cells, vascular endothelium and smooth muscle, and nasal gland. Strong immunoreactivity was shown in the mucosal epithelium of both groups, and weak to moderate reactivity in the mucosal epithelium of the inferior turbinate. Vascular endothelium and smooth muscle of both groups sometimes showed weak to moderate immunoreactivity. Nasal glands of both groups sometimes showed weak immunoreactivity. A significant difference between allergic and infectious groups was observed in predominant types of inflammatory cells. Neutrophils were predominant in the infectious group (p < 0.01), and eosinophils in the allergic group (p < 0.0001). About 50%-53% in allergic and 42% in infectious groups--of inflammatory cells showed positive immunoreactivity for iNOS. Immunoreactive cells were neutrophils, eosinophils, and macrophages. Lymphocytes, plasma cells, and mast cells invariably reacted negatively. A significant difference between allergic and infectious groups was observed in predominant iNOS-immunoreactive cells. Ratios of immunoreactive neutrophils to all neutrophils (p < 0.05) and to all inflammatory cells (p < 0.05) were significantly higher in the infectious group. The ratio of immunoreactive eosinophils to all inflammatory cells was significantly higher in the allergic group (p < 0.0001), while the ratio of immunoreactive eosinophils to all eosinophils did not differ between infectious and allergic groups. The ratios of immunoreactive macrophages to all macrophages and to all inflammatory cells did not differ significantly between groups. Electron microscopy showed that degenerated cells with pyknotic nuclei were located next to immunoreactive eosinophils, suggesting the cytotoxicity of NO, peroxynitrite, or superoxide.     

白细胞介素—6和8在鼻息肉组织中的表达

OBJECTIVE: To explore the role of interleukin-6,8 in nasal polyp formation and to search into the effect of allergy in the pathogenesis of nasal polyps (NP). METHODS: The expression and significance of interleukin-6,8 were studied in 36 nasal polyps and 36 serum samples of NP patients by radioimmunoassay. RESULTS: The mean value of IL-6 and IL-8 was (2.7658 +/- 0.3797) ng/L and (4.1877 +/- 0.1758) ng/L in all nasal polyp tissue homogenates. As compared with serum of NP patients, IL-6 and IL-8 were over expressed in nasal polyp tissue homogenates. No relation was found between the expression of IL-6/IL-8 and patients' gender, age and clinical stage. The expression of IL-6 and IL-8 in patients' serum, cord blood and normal serum showed no significant difference. CONCLUSION: IL-6 and IL-8 are strongly correlated with the formation of nasal polyp. Neither allergy nor infection play a role in the pathogenesis of nasal polyps.     

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。     

Exhaled inflammatory markers in aspirin-induced asthma syndrome

BACKGROUND: Interleukin (IL)-4 and IL-6, respectively, markers of neutrophilic and eosinophilic inflammation, were analyzed in nasal and oral exhaled breath condensate to understand the inflammation of upper and lower airways in subjects with aspirin-induced asthma (AIA) syndrome, evaluating possible differences between AIA and the single pathological conditions included in AIA syndrome. METHODS: Twelve patients with AIA, 17 patients with mild asthma (MA), 12 patients with nasal polyposis (NP), 11 patients with mild asthma + nasal polyposis (MA + NP), and 10 healthy subjects (HSs) were enrolled. Nasal and oral exhaled IL-4 and IL-6 were measured by enzyme immunoassay kit. RESULTS: Higher levels of nasal and oral exhaled IL-4 and IL-6 were observed in AIA compared with MA, NP, MA + NP, and HSs. Moreover, a positive correlation was identified between nasal exhaled IL-4 and IL-6 and, respectively, the number of neutrophils and eosinophils and in nasal scraping. CONCLUSION: The concentration of eosinophilic and neutrophilic markers in upper and lower airways of subjects with AIA syndrome is higher compared with HS and subjects with MA, NP, and MA + NP.     

鼻息肉中总IgE与嗜酸粒细胞浸润的相关性研究

目的:探讨总IgE及嗜酸粒细胞(EOS)在鼻息肉中的表达及其相关关系,以分析其在鼻息肉发病中的作用。方法:采用免疫组织化学SP法检测38例鼻息肉、15例中鼻甲黏膜组织中总IgE阳性细胞表达;采用Chromotrope2R染色法检测EOS浸润情况。结果:①鼻息肉组织中总IgE及EOS水平均明显高于中鼻甲黏膜组织,均P<0.01;②鼻息肉组织中总IgE与EOS呈明显正相关关系(r=0.843,P<0.01)。结论:EOS浸润在鼻息肉的形成和发展中起重要作用,IgE介导的Ⅰ型变态反应对鼻息肉中EOS浸润发挥一定作用。     

鼻息肉调节激活正常T细胞表达和分泌因子的测定及其意义

目的 探讨在鼻息肉形成过程中,上皮应答时产生调节激活正常Ｔ细胞表达和分泌因子（ｒｅｇｕｌａｔｅｄ ｕｐｏｎ ａｃｔｉｖａｔｉｏｎ,ｎｏｒｍａｌＴｃｅｌｌ ｅｘｐｒｅｓｓｅｄ ａｎｄ ｓｅｃｒｅｔｅｄ,ＲＡＮＴＥＳ）对嗜酸粒细胞趋化、移行、局部聚集的影响。方法 采用无血清原代细胞培养法培养鼾症患者下鼻甲上皮细胞和鼻息肉上皮细胞,经炎性介质ＩＬ－１β（２５ｕｇ／Ｌ,５０ｕｇ／Ｌ）刺激后收集２４ｈ和４８     

Inflammatory cells in nasal mucosa and nasal polyps

OBJECTIVE: Since some controversy exists concerning the frequency of inflammatory cells in nasal polyps, we have compared the frequency of tissue inflammatory cells (lymphocytes, neutrophils, eosinophils and plasma cells) including 11 kinds of lymphocyte subsets in the same specimens of nasal mucosa and nasal polyps. METHODS: Histopathological observations and flow cytometric analyses were performed on eight mucosal specimens of the inferior turbinates of patients with nasal polyps and on 13 polyp specimens. RESULTS: Nasal polyps contained significantly more eosinophils, neutrophils and plasma cells than nasal mucosa, and EG2+ cells (activated eosinophils) were significantly more frequent in nasal polyps than in nasal mucosa. Flow cytometric analysis showed that there were no significant differences in the frequencies of lymphocytes and lymphocyte subsets (CD1+, CD2+, CD3+, CD5+, CD7+, CD4+, CD8+, CD10+, CD19+, CD20+ and HLA-DR+ cells) including CD4/8 ratios between nasal mucosa and polyps, though, both nasal mucosa and polyps contained significantly more lymphocytes than eosinophils, neutrophils or plasma cells. The T cell lineage (CD2+, CD3+, CD5+ and CD7+ cells) was found in high frequency and B cell lineage (CD10+, CD19+ and CD20+ cells) in low frequency in both nasal mucosa and polyps. The frequency of HLA-DR+ cells (most of which were activated T cells) was not significantly different between nasal mucosa and nasal polyps. CONCLUSION: Histopathological and flow cytometric analyses were performed on the composition of inflammatory cells in nasal mucosa of the inferior turbinates and in polyps from the same patients. The elevated numbers of activated eosinophils, neutrophils and plasma cells in nasal polyps compared with nasal mucosa suggest that inflammatory processes play important roles in the pathophysiology of nasal polyps. The frequencies of lymphocytes and lymphocyte subsets were not significantly different between these two tissues.