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  • 1
    Electronic Resource
    Electronic Resource
    Cytokine gene expression and release from epithelial cells. A comparison study between healthy nasal mucosa and nasal polyps (1995)
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 25 (1995), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Backgroud Epithelial cells release cytokines and they probably contribute to chronic inflammation detected in bronchial asthma, rhinitis and nasal polyposis.Objectives To investigate the effect of cultures on cytokine gene expression to compare epithelial cell cytokine release by both healthy nasal nucosa (HNM) and nasal polyps (NP), and the modulation by dexamethasone and to investigate which cytokines may promote eosinophil survival.Methods Epithelials cells were cultured to confluence, human epithelial cell conditioned media generated with or without dexamethasone, and supernalanls measured by ELISA. Cytokine gene expression was investigated by reverse transcription-polymerase chain reaction (RT-PCR).Results Fresh epithelial cells only expressed mRNA for intesleukin-8 (IL-8) and granulocyte macrophage-colony stimulating factor (GM-CSF) while cultured cells expressed mRNA for IL-1β IL-6, IL-8, tumour necrosis factor-α (TNFα) and GM-CSF. Epithelial cells from NP significantly (P 〈 0.05) released more IL-8 (25431 ± 3163 pg/mL), and GM-CSF (1229 ± 391 pg/mL) than those from HNM (18604 ± 1723pg/mL for IL-8; and 611 ± 98 pg/mL for GM-CSF), Dexamethasone 10 μM inhibited the release of all cytokines, this effect being similar (40 50%) in both HNM and NP. except for IL-6 which was higher in HNM. Eosinophil survival induced by epithelial cell secretions from both HNM and NP was strongly blocked by GM-CSF antibody while it was partially blocked by antibodies to TNFα and IL-8.Conclusions These findings suggest that although epithelial cell culture procedures may upregulate cytokine gene expression, nasal polyps may represent a more active inflammatory tissue by releasing more cytokines than healthy nasal mucosa this release being inhibited by steroids; and that, in addition to GM-CSF.other cytokines such as TNFo and IL-8, may also be involved in the promotion of eosinophil survival.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2222.1995.tb01108.x
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  • 2
    Electronic Resource
    Electronic Resource
    Comparison of the role of nasal polyp and normal nasal mucosal epithelial cells on in vitro eosinophil survival. Mediation by GM–CSF and inhibition by dexamethasone (1994)
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 24 (1994), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Eosinophilic infiltration of the respiratory mucosa is considered an inflammatory hallmark of allergic rhinitis, bronchial asthma and nasal polyposis. However, the mechanisms involved in this infiltration have not yet been totally elucidated. The objective of this study was to investigate and compare the influence of epithelial cell secretions from both nasal polyps (NP) and normal nasal mucosa (NM) on in vitro eosinophil survival. Epithelial cells were identified by microscopy; and immunohislochemisiry. cultured to confluence, and human epithelial cell conditioned media (HECM) was generated from cultures. Eosinophits were isolated at high viability and purity (〉90%) from peripheral blood and incubated with HECM. HECM from both NM and NP increased eosinophil survival in a dose-dependent manner, this effect being maximal at a concentration of 25% for NM (73.4%±5.5%, n= 26, P〈 0.001) and of 10% for NP (74.5%± 84%n= 18, P 〈 0.001). Incubation of monoclonal antibody to human GM-CSF with HECM, neutralized the induction of eosinophil survival by HECM from both NM and NP. HECM from NP contained higher concentrations of GM-CSF (111 ± 25.4 pg/ml, n= 17) than HECM from NM (97.1 ± 15.2 pg/ml. n= 8). without reaching statistical significance. Pre-incubation of dexamethasone with eosinophils also blocked HFCM-induced eosinophil survival from both NM (10−8-10−5 M; IC50 = 9.5 nM) and NP (10-7-10-5 M; IC50 = 83 nM). These results suggest that: firstly eosinophil infiltration into the respiratory mucosa during allergic reaction and nasal polyposis may be modulated at least in part by GM-CSF from epithelial cells; and secondly epithelial cells from NP might have a more potent effect on inducing eosinophil infiltration of the respiratory mucosa than epithelial cells from NM. Finally, we may consider this as a reliable in vitro model to compare the role of epithelial cells from inflammatory (NP) and non-inflammatory (NM) tissue in respiratory inflammation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2222.1994.tb00240.x
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  • 3
    Electronic Resource
    Electronic Resource
    Effects of topical anti-inflammatory drugs on eosinophil survival primed by epithelial cells. Additive effect of glucocorticoids and nedocromil sodium (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 27 (1997), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Eosinophil infiltration is a hallmark of the inflammatory response in rhinitis and in nasal polypcsis.Objective We studied the effect of steroids and nedocromil sodium on eosinophil survival primed by epithelial cells from healthy (nasal mucosa) and inflamed (nasal polyp) respiratory tissue.Methods Blood eosinophils were incubated with increasing concentrations (10-11 10-5 M) of topical steroids (fiuticasone propionate, budesonide, triamcinolone acetonide and beclomethasone dipropionate) and/or nedocromil sodium prior to the addition of human epithelial cell conditioned media (HECM), eosinophil viability was measured and IC50 for each drug was calculated.Results All four steroids and nedocromil sodium caused a dose-related inhibition of HECM-induced eosinophil survival. The IC50 of steroids were lower in eosinophils primed by mucosa HECM than on those primed by polyp HECM (fluticasone, 4nM vs 114nM: budesonide, 21 nM vs 280 nM; triamcinolone, 7 nM vs 853 nM; and beclomethasone, 171 nM vs 181 nM). The combined inhibitory effect of 10-7M budesonide plus 10-5M nedocromil (43.8 ± 10.8%, P 〈 0.03) was significantly higher than budesonide (28.5 ± 9.2%) or nedocromil (16.7 ± 5.4%) alone and close to 10-5M budesonide (52.3 ± 11%). No differences were found in cytokine (IL-8, IL-6, GM-CSF, TNFα, IL-lβ and RANTES) concentrations between HECM from mucosa and polyps.Conclusion These results suggest that topical anti-inflammatory drugs may diminish airway eosinophilic infiltration by decreasing eosinophil viability, that nasal polyp epithelial cell secretions may induce steroid resistance in eosinophils, and that nedocromil sodium has additive effects with steroids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2222.1997.1750975.x
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  • 4
    Electronic Resource
    Electronic Resource
    Identification of mast cells in bronchoalveolar lavage fluid (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 46 (1991), S. 0 
    Details
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The present study was carried out to compare the effectiveness of different fixation and staining methods in the identification of mast cells obtained by bronchoalveolar lavage from patients with interstitial lung diseases. Cell preparations were fixed with formaldehyde or methanol. Mast cells were identified by metachromatic staining with May Grunwald Giemsa, Toluidine blue or Gallamine blue Giemsa. After formaldehyde fixation only a few mast cells were identified, regardless of the stain method used. In contrast, a significantly higher number of mast cells were observed after methanol fixation. Using this fixative, Toluidine blue stain showed a higher number of mast cells than May Grunwald Giemsa. However, there was no difference between Toluidine and Gallamine blue Giemsa in the number of cells observed. The easy identification of mast cells after staining with toluidine, combined with its easy application, suggest that Toluidine blue stain after methanol fixation is the most useful method for determining the presence of mast cells in lavage fluid.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1398-9995.1991.tb00575.x
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  • 5
    Electronic Resource
    Electronic Resource
    Immunocytologic analysis of nasal cells obtained by nasal lavage: a comparative study with a standard method of cell identification (1993)
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 48 (1993), S. 0 
    Details
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: For evaluation of two methods of nasal cell identification, cell morphology and immunocytologic analysis, nasal lavage was performed in 16 healthy subjects and 29 patients suffering from rhinitis. Nasal lavage smears were stained with May-Grünwald-Giemsa (MGG), and cells were identified according to their structure as epithelial cells, neutrophils, lymphocytes, eosinophils, and metachromatic cells (basophils and mast cells). Immunocytologic analysis was performed with monoclonal antibodies by the immunoalkaline phosphatase method. The following monoclonal antibodies were used: CK1, EG2, and CD3, which identify epithelial cells, activated eosinophils, and T lymphocytes, respectively; CD 15, which recognizes mature granulocytic cells; and CD 14, which reacts with monocytes and macrophages. A significant difference was observed between the two methods in the number of identified epithelial cells, in both controls (64.6 ± 7.8% with MGG, 14.2 ± 3.5% with CK1 analysis) and patients with rhinitis (56.9 ± 7.6% with MGG, 18.2 ± 3.7% with CK1 analysis). In contrast, no significant differences were found in eosinophil and neutrophil counts when the two methods were compared. After nasal allergic provocation, a significant increase in the number of eosinophils was observed with both methods in seven patients with rhinitis. The results of this study indicate that: 1) MGG staining is a useful method to identify the cells obtained by nasal lavage, and 2) immunocytologic analysis with monoclonal antibodies accurately identifies granulocytic cells, while only a low proportion of epithelial cells are detected, probably because anticytokeratin monoclonal antibody reacts only with viable cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1398-9995.1993.tb00753.x
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