Ultrastructure of mast cells in nasal polyps]

More mast cells were found in the base of nasal polyps and most of them showed varying degrees of degranulation. There were three types of granules in the cytoplasm, namely: high electron-dense granules with crystalline structure; low electron-dense granules with netlike structure and some granules containing a mixture of components of different electron density. The appearance of cell degeneration in nasal polyps and granules broken away from mast cells through the dissolved cellular membrane were noted. These findings suggested that mast cells degranulation in nasal polyps might be in some way different from the more rapid changes found in cells activated by the classical IgE-mediated pathway and showed that mast cell degranulation may play a part in formation and growth of the nasal polyps.     

Effects of glucocorticoids on infiltrating cells and epithelial cells of nasal polyps

Glucocorticoids are known to be effective in the treatment of nasal polyps (NPs). To examine the mechanisms of their effect, we evaluated 1) the ability of glucocorticoids to induce the apoptosis of eosinophils and T lymphocytes in NPs, and 2) the ability of dexamethasone to down-regulate epithelial cell functions that relate to eosinophilic inflammation. In vitro and in vivo, glucocorticoids increased the apoptosis of both eosinophils and T lymphocytes in NPs. Dexamethasone inhibited the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) from both NP epithelial cells that were unstimulated and NP epithelial cells that were stimulated with interleukin-4 or tumor necrosis factor alpha. These results suggest that the clinical efficacy of glucocorticoids on NPs may be due to 1) induction of apoptosis in both eosinophils and T lymphocytes that infiltrate NPs, and 2) down-regulation of epithelial GM-CSF production, which prolongs eosinophil survival.     

肥大细胞在鼻息肉中分布特点分析

目的：探讨肥大细胞（MC）在鼻息肉中的分布特点及向上皮层移动的机制。方法：以免疫组织化学染色方法观察鼻息肉组织中MC的分布情况,观察CCL5、CCL11、CX3CL1、IL-8、IL-6等MC趋化因子在鼻息肉中的表达情况。结果：MC在鼻息肉组织中向上皮层移动,CCL5、CCL11、CX3CL1、IL-8在鼻息肉组织上皮层中的表达均明显增强;但在鼻息肉上皮层未见IL-6免疫阳性细胞,在间质的炎性细胞中偶见有免疫阳性细胞。结论：MC在鼻息肉中向上皮层移动,其移动的机制可能是上皮细胞受炎症因素刺激后CCL5、CCL11、CX3CL1、IL-8、IL-6等MC趋化因子的表达增加的原因。MC在鼻息肉的形成过程中可能发挥重要作用。     

Regulation of immunoglobulin secretion by plasma cells infiltrating nasal polyps

OBJECTIVE/HYPOTHESIS: To learn more about the role of plasma cells infiltrating nasal polyps in the pathogenesis of nasal polyposis, we examined their function by analyzing immunoglobulin (Ig) production and the factors implicated in the secretion. STUDY DESIGN: A series of 19 consecutive nasal polyp tissue samples and, as a control, peripheral blood samples from the same patients, were studied by histopathological and immunological examination. METHODS: Hematoxylin-eosin and immunohistochemical staining was carried out to identify plasma cells infiltrating nasal polyps. Nasal polyp mononuclear cells (NPMNCs) were purified from nasal polyp tissue samples, and Ig-secreting cells were identified in cytospin preparations stained with fluorescein isothiocyanate-conjugated antibodies against IgA, IgG, IgM, and IgE. Purified NPMNCs were cultured in basal conditions and after the addition of several stimuli. Ig secreted into the culture supernatants was evaluated by an enzyme-linked immunosorbent assay. RESULTS: Plasma cells accounted for an important fraction of the inflammatory infiltrate. The main Ig isotype synthesized by these cells was IgA, whereas little IgE was detected. In vitro cultures demonstrated that the plasma cells actively secreted Ig for a short period. When cytokine dependence was analyzed, interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-alpha) were shown to be partially responsible for the Ig production. Dependence on CD95-mediated apoptosis was not observed. CONCLUSIONS: Nasal polyp-infiltrating plasma cells are mainly IgA-secreting cells, the latter property being related to the mucosal immune system. The IgA production is partly dependent on IL-10 and TNF-alpha. The absence of IgE-secreting cells in most of the samples suggests that a type I hypersensitivity reaction is not essential for the development of nasal polyp.     

Role of mast and goblet cells in the pathogenesis of nasal polyps

In this study, the role of mast and goblet cells and eosinophils in the pathogenesis of nasal polyposis was investigated. The study group consisted of 28 adult patients (15 males, 13 females) with nasal polyposis who underwent functional endoscopic sinus surgery (FESS). All patients in the study group were examined with a questionnaire, an otolaryngologic examination, an endoscopic examination with 0 degrees and 30 degrees endoscopes, Waters' graphy, and axial and coronal computed tomography of the paranasal sinuses. The control group consisted of 10 adult patients without nasal polyp (7 males and 3 females) who underwent septoplasty. They gave written approval to enter the study. The polyp specimens from the study group were excised from four regions: the maxillary sinus, ethmoid sinus, sphenoid sinus, and nasal cavity. They were examined at x400 magnification by light microscopy, and only the slides with polypoid tissue were included in the study. Slides including a chronic inflammatory process without polypoid tissue were excluded from the study. The control group was composed of the slides of specimens from the inferior turbinate. Forty slides (10 in each group) in the study group and 10 slides in the control group were included in the study. The surgical specimens from the study and control groups were examined with a histochemical staining technique. In every surgical specimen, the type of epithelium and the numbers of goblet and mast cells and eosinophils were calculated in x400 high-magnification field in 10 areas on light microscopy, as well as the mean number of these cells, and for mast cells separately, cell count in the epithelium and the stromal layer of polyp tissue and total mast cell count, including both epithelial and stromal mast cells, were identified. Goblet cells, mast cells, and inflammation with eosinophils were observed in all sinonasal mucosa. The common epithelial type in the polyp tissue was pseudostratified ciliated cylindric epithelium, which contains goblet cells. Goblet cell numbers in the maxillary, ethmoid, and sphenoid sinuses and nasal cavity were found to be significantly higher than in the control group (p < .05). For total mast cell and eosinophil count, no statistically significant difference was found between all five groups. In each group, there was no statistically significant difference between goblet and mast cells. Increased goblet cells in sinonasal polyps indicated that systemic factors also affect nasal polyposis as much as local factors, such as airflow and mucosal contact. Surgical treatment of sinonasal polyps by FESS causes more sufficient air ventilation in the nasal cavity and paranasal sinuses. Therefore, the goblet cell density will decrease because of the exposure of the mucosal surfaces to the air. In particular, FESS and then the appropriate medical treatment may decrease the recurrence rates and increase the patient's comfort. The significantly increased goblet cell count in the sinonasal mucosa demonstrated the importance of these cells in the pathogenesis of nasal polyposis. Also, mast cells and eosinophils may have a role in the inflammatory processes, leading to nasal polyposis formation.     

Differentiation and proliferation of mast cells in nasal mucosa]

We determined the distribution of mast cells in nasal mucosa and studied their proliferation. Inferior turbinate mucosa was sampled in 13 patients with allergic rhinitis (allergic group) and 5 without (non allergic group) and stained immunohistochemically using anti mast cell tryptase antibody, anti-c-kit antibody, anti-PCNA antibody, and anti mast cell chymase antibody. Tissue sections stained with anti tryptase antibody revealed a higher degree of infiltration of tryptase-positive cells, i.e., mast cells, in the allergic group than in non allergic group. In the allergic group, the number of tryptase-positive cells, c-kit-positive cells, and PCNA-positive cells was significantly greater in the epithelium and shallow lamina propria than that in the deep lamina propria. Tryptase-positive, c-kit-positive cells, i.e., c-kit-positive mast cells, and tryptase-positive, PCNA-positive cells, i.e., PCNA-positive mast cells, were also abundunt in the epithelium and shallow lamina propria. The stem cell factor and c-kit receptor are reported to play a primary role in mast cell differentiation and proliferation. PCNA-positive cells represent actively proliferating cells. Based on the above, we concluded that mast cells in the epithelium and shallow lamina propria in the allergic group differentiated and proliferated more actively than those in the deep lamina propria.     

The effects of different fixations on the distribution and numbers of mast cells in patients with nasal polyps

Mast cells in rats have been categorized into mucosal and connective tissue mast cells by properties which include their fixation characteristics and their distribution in tissues. There are a number of factors which make this classification unsatisfactory including the variability of fixation, intensity of background staining and the application of animal studies to man. Seasonal allergic reactions in the nose of man occur both in the surface metachromatic cells and in those within the epithelium; the latter are presumed to be mucosal mast cells. The aim of this study was to look at the distribution and the fixation characteristics of mast cells in inferior turbinates and nasal polyps. Both techniques show that cells are more abundant in the submucosa and the fact that Carnoy's fixative shows them better and this may be due to the lack of intensity in the background staining. There is no evidence that mast cells are more frequent in the epithelium. These findings would suggest the nasal polyps are not an allergic disease.     

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.     

Mucociliary function in patients with nasal polyps

Patients with nasal polyposis complain of nasal blockage and rhinorrhoea, which may be due to impaired mucociliary clearance. The saccharine clearance time and ciliary beat frequency of samples of ciliated epithelium from patients with nasal polyps was measured. We also studied the effect of fluid from the oedematous stoma of nasal polyps and of histamine and prostaglandin (PG) D₂ E₂ and F_2x on the cilia from normal individuals. Polyp fluid was found to increase ciliary beat frequency. Histamine and PGD2 had no effect, but PGE₂ and PGF_2x both increased ciliary beat frequency and so may cause the ciliostimulatory effect of polyp fluid. The saccharine clearance time was prolonged in three of nine patients, but ciliary beat frequency was only slightly reduced in one of these. Thus, where mucociliary clearance is reduced, it is likely to be due to abnormalities of mucus rather than impaired ciliary activity.     

鼻息肉上皮细胞的分化

目的 探讨鼻息肉上皮细胞的分化状态及其病理学意义.方法 采用免疫组化染色法检测鼻息肉组(29例)和下鼻甲组(11例)上皮细胞CK14、CK18 和CK13蛋白的表达.结果 ①鼻息肉组上皮细胞CK14阳性指数,CK18阳性指数,CK13区域性表达率均大于下鼻甲组(P<0.01).②鼻息肉上皮下微腺体上皮细胞CK18 高表达,并与鼻息肉上皮CK18 阳性细胞呈连续渐变的显微解剖学关系.结论 ①鼻息肉上皮基底细胞具有多向分化潜能,可以分化为假复层纤毛柱状上皮、非角化复层鳞状上皮和间质腺上皮,形成鼻息肉上皮种类的多样性.②鼻息肉鳞状化生现象较为多见,这可能是鼻息肉合并内翻性乳头状瘤和鼻息肉上皮存在原位癌区的病理学基础.     

Hypoxia effects on vascular endothelial growth factor derived epithelial cells of nasal polyps]

OBJECTIVE: To understand the role of nasal mucous epithelial cells to hypoxia in early stage of nasal polyps(NP) formation. METHODS: Epithelial cells of NP and inferior turbinate (IT) were cultured without serum under normal oxygen and hypoxia, and stimulus of inflammatory cytokines. Erythropoietin (EPO) was regarded as hypoxia mark, and expression of vascular endothelial growth factor(VEGF) mRNA and protein derived from epithelial cells were detected respectively by in situ hybridization and ELISA. RESULTS: 1. Under hypoxia, EPO mRNA was expressed intensely in epithelial cells from NP and IT, and there was no significant difference between both of them. This result suggested that EPO might be regarded as a hypoxic mark. 2. The ability of producing VEGF mRNA increased with cytokines stimulation, especially under hypoxia. Protein level of VEGF from epithelial cells of NP and IT increased with cytokines stimulation, especially in hypoxia and was time-dependent. CONCLUSION: Epithelial cells actively produce vast VEGF under hypoxia. The VEGF induced by hypoxia of the mucosa in middle meatus is of importance in the formation of nasal polyps(NP) in early stage, which may be the major cause of NP formation in middle meatus.     

鼻息肉及变应性鼻炎中IL-17表达与嗜酸粒细胞浸润的意义

目的:观察鼻息肉与变应性鼻炎中IL-17表达与嗜酸粒细胞(Eos)浸润情况,探讨IL-17与Eos在其发病中作用及相关性。方法:采用免疫组织化学SP法检测鼻息肉21例、变应性鼻炎18例及单纯中隔偏曲12例中IL-17表达情况,苏木精-伊红染色下观察炎性细胞浸润并计数Eos进行比较分析。结果:①发现在变应性鼻炎鼻黏膜下固有层、鼻息肉组织上皮层及间质内Eos、中性粒细胞为主的炎性细胞及部分腺体内有IL-17表达;②检测3组中免疫组织化学IL-17阳性细胞数、IOD(IL-17阳性表达吸收度值)及苏木精-伊红染色Eos计数,发现鼻息肉与变应性鼻炎间差异无统计学意义(P>0.05),但均高于正常对照组,差异有统计学意义(P<0.05)。Eos数与IL-17阳性细胞数在鼻息肉中存在正相关性,相关系数r=0.606,P<0.01,在变应性鼻炎中两者亦有相关性,相关系数R=0.446,P<0.05。结论:①IL-17是一种新型的细胞因子,在鼻息肉组织与变应性鼻炎鼻黏膜组织中有表达,因此在发病过程中可能起调节免疫炎症反应作用,可做为深入研究鼻息肉与变应性鼻炎发病机制的指标;②推断在鼻息肉及变应性鼻炎发病中IL-17亦有趋化E...     

两种细胞外基质糖蛋白在鼻息肉中的表达

OBJECTIVE: To explore the role of tenascin (TN) and fibronectin (FN) in the pathophysiology of nasal polyp. METHODS: The expression of TN and FN in nasal polyps from 34 patients and in inferior turbinates from 20 patients with deviation of nasal septum was studied with immunohistochemical method. In patients with nasal polyps, the relations between expression and histopathologic characteristics, eosinophilias (EOS) infiltration, clinical staging and the size of nasal polyps were analyzed. RESULTS: (1) The gray score of TN and FN expression was 163.10 +/- 10.54 and 163.24 +/- 11.52 in nasal polyps respectively, whereas it was 175.49 +/- 9.29 and 173.93 +/- 7.92 in inferior turbinates respectively. The difference between two groups was significant(P < 0.01); (2) The expression of TN and FN in edematous type was significantly stronger than that in cystic fibrous and glandular type (P < 0.05); (3) The association between FN expression and EOS infiltration was significant(r = -0.60, P < 0.01); (4) The expression of TN and FN did not correlate with clinical staging and size(P > 0.05). CONCLUSION: Abnormal ECM may contribute to proliferation of epithelium, accumulation of EOS and edema formation, as a result, to enhance the development of nasal polyps.     

Expression and distribution of Aquaporin 1 in the nasal polyps]

OBJECTIVE: To confirm the expression and distribution of Aquaporin 1 (AQP1) in the nasal polyps and to investigate the relation between AQP1 and the nasal polyp edema. METHODS: fourteen cases of normal inferior turbinates and 26 cases of nasal polyps were used. The expression and distribution of AQP1 in nasal polyps were examined by immunohistochemical technique. RESULTS: The level of AQP1 in the epithelial cells and serous cells from nasal polyps was higher than that in inferior turbinates, and the level of AQP1 in the epithelial cells and cilium cells from inferior turbinate was higher than that in nasal polyps. CONCLUSION: There is close relationship between AQP1 and nasal polyp edema.     

Effect of proinflammatory cytokines on human leucocyte antigen expression of epithelial cells of nasal polyps]

OBJECTIVE: To explore the effect of proinflammatory cytokines on the expression of human leucocyte antigen(HLA-DR) in epithelial cell of nasal polyps. METHODS: The samples were from 23 patients with nasal polyps and 11 cases of normal inferior turbinates. The samples were fixed, paraffin-embeded and sectioned for observing distribution of HLA-DR positive cells. Nasal epithelial cells were obtained after enzymatic digestion and cell culture was performed for observing influences of interleukin (IL)-1 beta and tumor necrosis factor (TNF-alpha) to expression of HLA-DR of epithelial cells and the effect of dexamethasone(10(-5) mol/L). RESULTS: The HLA-DR expression in epithelium was significantly stronger in nasal polyps than that in inferior turbinates. The expression of HLA-DR in cultured cells under stimulation of proinflammatory cytokines was increased compared with controls, but the expression was more intense in nasal polyps than that in inferior turbinates. Increased HLA-DR expression under stimulation of cytokines was reduced significantly by dexamethasone. CONCLUSION: The epithelium is in a key position to participate in inflammatory and immunologic events in the airways. Epithelial cells potentially contribute to airway inflammation by antigen presentation and the production of proinflammatory cytokines.     

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     

嗜酸粒细胞主体碱性蛋白对鼻粘膜上皮肥大细胞组释放的影响

Eosinophils and nasal epithelial mast cells (NEMCs) are closely related in nasal allergy. The activity of the eosinophil cationic protein, major basic protein (MBP) on histamine release from human NEMCs has not been documented. We examined the effects of MBP on histamine release from NEMCs. Nasal scrapings (containing NEMCs) from 59 patients with house dust mite nasal allergy were incubated with MBP or alkylated MBP. Specimens were also pre-incubated with MBP followed by incubation with house dust mite allergen (HDM) or preincubated with HDM followed by incubation with MBP or HDM. Histamine released in incubated media and remained in incubated mast cells was quantified by radio-immunoassay. MBP at the concentration of 10(-5) mol/L induced mild but statistically significant histamine release from the NEMCs. Preincubation with MBP at the concentration of 1.57 x 10(-5) mol/L produced slight inhibition of subsequent HDM-induced histamine release but preincubation with HDM showed no effect on the subsequent MBP-induced or HDM-induced histamine release from NEMCs. These results suggest that accumulated and activated eosinophils may release MBP resulting in clinical symptoms by interaction with NEMCs and inhibit of histamine release from NEMCs by subsequent interaction with allergen.