Nerve growth factor localization in the nasal mucosa of patients with persistent allergic rhinitis

Background and objectives:  Nerve growth factor (NGF) and NGF receptors have been shown to be expressed by structural and infiltrating inflammatory cells in the human allergic bronchial mucosa and conjunctiva. In the nose, a positive immunostaining for NGF was recently reported in biopsies of subjects undergoing surgery for refractory nasal obstruction. This study was aimed at studying by immunohistochemistry NGF expression and localization in the nasal mucosa from subjects with moderate/severe persistent allergic rhinitis and natural allergen exposure.
Methods:  Immunostaining for NGF, tryptase and eosinophil cationic protein was performed in human nasal turbinate sections of 25 patients affected by persistent allergic rhinitis and sensitization to Dermatophagoides pteronyssinus .
Results:  NGF was consistently expressed in the epithelium and in the submucosa of allergic rhinitic subjects, preferentially localized in eosinophils and mast cells. A strong NGF immunostaining was found in mucous cells of the epithelial lining and in the submucosal glands.
Conclusions:  As previously shown for allergic asthma and allergic conjunctivitis, NGF is also detectable in the nasal mucosa of patients with persistent allergic rhinitis. The preferential NGF localization in mucous cells of the epithelial lining and in the submucosal glands suggests a possible role for NGF in modulating secretion in allergic rhinitis and possibly other allergic diseases.     

Surgical treatment of the inferior turbinate for allergic rhinitis: clinical evaluation and therapeutic mechanisms of the different techniques

Many different surgical procedures have been described to alleviate nasal obstruction due to turbinate hypertrophy in patients with allergic rhinitis (AR) who continuously have chronic nasal congestion. The goal of surgical procedures should be 'optimal reduction of volume and secretion with preservation of function'. For a period of about 25 years, laser surgery of the inferior turbinate has been considered a simple and effective surgical treatment with a low complication rate that has been in widespread use by many ENT surgeons. Various trials have demonstrated the clinical effectiveness of a series of different laser systems including carbon dioxide, Nd:YAG, potassium-titanyl-phosphate, and diode laser. Laser surgery generally induces significant increases of the total volume of the nasal cavity. Several studies have provided evidence that the expression degree of local inflammatory cytokines and chemical mediators can be attenuated by the treatment. The mucociliary function of the inferior turbinate seems to be preserved after CO₂ laser intervention. A breakthrough in the surgical procedures available for allergic patients who severely suffer from both intractable hypertrophic inferior turbinates and unregulated nasal secretion was introduced in Japan. Posterior nasal neurectomy is a novel alternative method in which neural bundles are selectively cut or cauterized from the sphenopalatine foramen. Resection of the posterior nasal nerve at this point enables surgeons to inhibit the parasympathetic nerve function of the nasal mucosa with no troublesome complications such as decrease of lacrimal secretion. The procedure simultaneously provides interruption of the somatic afferent innervation to the turbinate mucosa. Herein we assess the clinical effectiveness of this surgical procedure based on objective and measurable monitoring parameters and discuss the underlying therapeutic mechanisms.     

Histaminergic Regulation of Natural Killer Cell-Mediated Clearance of Tumour Cells in Mice

Treatment of Swiss albino mice with histamine enhanced the clearance of natural killer (NK)-cell sensitive YAC-1 lymphoma and B16/F10 melanoma cells from lung tissue in vivo , but did not affect the elimination of NK-cell-insensitive P815mastocytoma cells. The effect of histamine was apparently mediated by H₂-type histamine receptors (H₂R) since it was blocked by ranitidine, an H₂R antagonist. Histamine did not affect clearance of tumour cells in animalsdepleted of NK cells in vivo by treatment with antibodies to asialo-GM1 or NK1.1. The effect of histamine was time-dependent: pretreatment with histamine for 3 h significantly augmented the clearance of YAC-1 cells, whereas, pretreatmentwith histamine for 5 min was ineffective. Histamine potentiated the anti-tumour properties of NK-cell activators such as interleukin-2 (IL-2) or interferon-α (IFN-α) in vivo . None of these lymphokines significantly affected theclearance of YAC-1 cells unless animals were concomitantly treated with histamine. Treatment with ranitidine alone reduced the in vivo clearance of YAC-1 cells from lungs but did not affect the clearance of NK-cell-insensitive P815 cells. Effects of ranitidine on NK-cell function in vivo were not shared by a chemical control to ranitidine, AH20239AA, thus indicating that the inhibition of NK-cells results from H₂R antagonism rather than non-specific toxicity. It is concludedthat histaminergic mechanisms may be involved in the regulation of NK cell function in vivo     

Book reviews

Book reviewed in this article:
H₁ and H₂ Histamine Receptors edited by Guy A. Settipane     

Liquid secretion properties of airway submucosal glands

The tracheobronchial submucosal glands secrete liquid that is important for hydrating airway surfaces, supporting mucociliary transport, and serving as a fluid matrix for numerous secreted macromolecules including the gel-forming mucins. This review details the essential structural elements of airway glands and summarizes what is currently known regarding the ion transport processes responsible for producing the liquid component of gland secretion. Liquid secretion most likely arises from serous cells and is principally under neural control with muscarinic agonists, substance P, and vasoactive intestinal peptide (VIP) functioning as effective secretogogues. Liquid secretion is driven by the active transepithelial secretion of both Cl⁻ and HCO₃⁻ and at least a portion of this process is mediated by the cystic fibrosis transmembrane conductance regulator (CFTR), which is highly expressed in glands. The potential role of submucosal glands in cystic fibrosis lung disease is discussed.     

Histamine content, synthesis and degradation in human nasal mucosa

Histamine content and enzyme activities of histamine metabolism, histidine decarboxy-lase (HDC), histamine N-methyltransferase (HMT) and histaminase (diamine oxidase, DAO) in human nasal mucosa were determined with a highly sensitive and specific fluorescent method which was combined with high performance liquid chromatography. Histamine content and HDC activity were determined in 10 specimens of nasal polyp, nine specimens of maxillary sinus and five specimens of inferior turbinate. HMT and histaminase activities were determined in 15 specimens of nasal polyp, nine specimens of maxillary sinus and five specimens of inferior turbinate obtained during surgical therapy. Histamine and activities of HDC, HMT and histaminase were detected in all specimens except the case of histaminase activity in one specimen of nasal polyp. The mean values of histamine content and activities of HDC, HMT and histaminase of human nasal mucosa were 137.3 nmol/g wet weight, 26.3 fmol/min/mg protein, 26.4 pmol/min/mg protein and 0.5 pmol/min/mg protein, respectively. Histamine content in the mucosal tissue of the maxillary sinuses was significantly higher than that of nasal polyps or inferior turbinates. There were no significant differences in HDC activities among three kinds of nasal mucosa. Activities of HMT and histaminase, including their kinetic constants (Km and Vmax values for histamine) indicated that HMT has a greater potential than histaminase for histamine degradation in the human nasal mucosa. The presence of these enzymes suggests that these activities constitute an important modulating factor in histamine mediated allergic and inflammatory reactions in human nasal mucosa.     

Characterization and autoradiographic localization of histamine H1 receptors in human nasal turbinates.

To examine the localization of histamine H1 receptors (H1R) in human nasal mucosa, the autoradiographic distribution of H1R was studied in human nasal inferior turbinates. Cryostat sections were incubated with various concentration of [3H]pyrilamine in saturation-binding studies and with 1 nmol/L of [3H]pyrilamine for autoradiography. Nonspecific binding was determined by adding 2 mumol/L of pyrilamine. Scatchard analysis demonstrated high-affinity binding sites with a maximum binding capacity of H1R of 193 +/- 46 fmol/mg of protein, and dissociation constant was 0.6 +/- 0.1 nmol/L. Autoradiograms indicated H1R exist exclusively on the endothelium of vessels. No specific labeling could be observed in the submucosal glands or epithelium. These results extend and support our previous finding that histamine directly causes vascular permeability through H1R and stimulates nasal glandular secretion indirectly through reflexes.     

Interleukin-6 is essential for Staphylococcal exotoxin B-induced T regulatory cell insufficiency in nasal polyps

Background The pathogenesis of nasal polyps is still unclear. There is increasing evidence indicating that Staphylococcal aureus (S. aureus) is associated with the formation of nasal polyps, but the mechanism has not been well documented to date.
Methods We stimulated cultured nasal polyps and turbinate tissues with Staphylococcal exotoxin B (SEB), detected the expression of pro-inflammatory cytokines (IL-2, IL-6, and IL-8) and T cell cytokines (IFN-γ, IL-4, IL-5, IL-10, and IL-17) in the supernatants, and evaluated mRNA expression (T-bet, GATA-3, Foxp3, and RORγt) and frequencies of CD4⁺CD25⁺ T regulatory cells (Tregs) in nasal tissues. We also evaluated the effects of blocking IL-6 with monoclonal antibodies to T cell profiles in cultured nasal tissues stimulated by SEB.
Results Levels of IL-6, IFN-γ and IL-4 increased significantly in SEB-stimulated nasal polyps. Meanwhile, mRNA expressions of T-bet and GATA-3 were significantly up-regulated, while Foxp3 was inhibited and the frequencies of CD4⁺CD25⁺ Tregs were decreased after SEB stimulation. After blocking IL-6, the levels of IL-10 and Foxp3 mRNA, as well as the frequencies of CD4⁺CD25⁺ Tregs, were significantly increased, while IFN-γ and IL-4 production and the mRNA expression of T-bet and GATA-3 were significantly inhibited.
Conclusions SEB is able to modulate pro-inflammatory factors, T-helper type 1/Th2 profiles and suppress Treg activity in cultured nasal polyps, which were rescued by blocking IL-6 activity. Therefore, IL-6 is essential for SEB-induced Treg insufficiency in nasal polyps.     

Forkhead box P3+ T cells express interleukin-17 in nasal mucosa of patients with both allergic rhinitis and polyposis

The pathogenesis of nasal polyposis remains unclear; it severely affects patients' quality of life and complicates inflammation in adjacent organs such as sinusitis and asthma. Aberrant immune regulatory function in these patients is proposed. The present study aims to examine the regulatory T cells (T(reg) ) in nasal mucosa of patients with allergic rhinitis (AR) and nasal polyposis (NP). Patients with AR or AR/NP were treated with inferior turbinectomy for their inferior turbinate hyperplasia. Surgically removed nasal mucosa was collected to examine the T(reg) by immunohistochemistry and flow cytometry. The results showed that more forkhead box P3 (FoxP3)(+) cells were found in AR with polyps than in those with AR alone. Further studies revealed that these FoxP3(+) T cells from AR/NP group also expressed interleukin (IL)-17. In vitro study showed that staphylococcal enterotoxin B (SEB) induced CD4(+) FoxP3(+) T cells to become FoxP3(+) IL-17(+) cells via facilitating the expression of IL-6, that in synergy with transforming growth factor-beta, induce the expression of IL-17 in FoxP3(+) cells. We conclude that FoxP3(+) IL-17(+) T cells were localized in the nasal mucosa of patients with AR and NP. SEB may play a role in converting FoxP3(+) T(reg) to FoxP3(+) IL-17(+) T cells. The presence of IL-17(+) FoxP3(+) T cells may play a role in the remodelling of the nasal airways in certain people who develop polyps, irrespective of whether or not they are atopic.     

Histaminergic Regulation of NK-Cells: Protection Against Monocyte-Induced Apoptosis

Human natural killer (NK) cells (with CD3⁻/56⁺ phenotype) acquired features characteristic of apoptosis after incubation with autologous monocytes, as revealed by apoptotic nuclear morphology and degradation of DNA into oligonucleosomal fragments. The monocyte-induced apoptosis in NK-cells was prevented by the biogenic amine histamine at concentrations exceeding 0.1 μ M . The protective effect of histamine was blocked by the H₂-receptor (H₂R) antagonist ranitidine but not by AH202399 A, a chemical control to ranitidine devoid of H₂R affinity. It is concluded that histaminergic mechanisms may serve to protect NK cells from damage inflicted by products of the oxidative metabolism of monocytes.     

IP3 receptors: some lessons from DT40 cells

Summary:  Inositol-1,4,5-trisphosphate receptors (IP₃Rs) are intracellular Ca²⁺ channels that are regulated by IP₃ and Ca²⁺ and are modulated by many additional signals. These properties allow them to initiate and, via Ca²⁺-induced Ca²⁺ release, regeneratively propagate Ca²⁺ signals evoked by receptors that stimulate formation of IP₃. The ubiquitous expression of IP₃R highlights their importance, but it also presents problems when attempting to resolve the behavior of defined IP₃R. DT40 cells are a pre-B-lymphocyte cell line in which high rates of homologous recombination afford unrivalled opportunities to disrupt endogenous genes. DT40-knockout cells with both alleles of each of the three IP₃R genes disrupted provide the only null-background for analysis of homogenous recombinant IP₃R. We review the properties of DT40 cells and consider three areas where they have contributed to understanding IP₃R behavior. Patch-clamp recording from the nuclear envelope and Ca²⁺ release from intracellular stores loaded with a low-affinity Ca²⁺ indicator address the mechanisms leading to activation of IP₃R. We show that IP₃ causes intracellular IP₃R to cluster and re-tune their responses to IP₃ and Ca²⁺, better equipping them to mediate regenerative Ca²⁺ signals. Finally, we show that DT40 cells reliably count very few IP₃R into the plasma membrane, where they mediate about half the Ca²⁺ entry evoked by the B-cell antigen receptor.     

Involvement of inferior turbinate mucosa in chronic sinusitis--localization of T-cell subset

BACKGROUND: In chronic sinusitis (CS), different subsets of leukocytes are involved in development of persistent inflammation of the nasal mucosa. The localization and differentiation of these infiltrating lymphocytes may help us to understand the inflammatory interactions in the epithelium, lamina propria, and seromucous glands of the nasal mucosa in CS. METHODS: We examined frozen sections of inferior turbinates from 14 patients with nonallergic CS and 10 normal nonallergic controls. We used the avidin-biotin-peroxidase (ABC) technique with monoclonal antibodies against CD3 (total T cells), CD4 (T-helper/inducer cells), CD8 (T-suppressor/cytotoxic cells), CD22 (B cells), CD56 (natural killer cells), elastase (neutrophil granulocytes), eosinophil cationic protein (eosinophil granulocytes), and CD68 (macrophages). RESULTS: We found significant increases (P < 0.05) of CD3, CD4, and CD8 T cells and B cells in the nasal mucosa of patients with CS. The number of CD68 cells and eosinophils showed no significant rise. CONCLUSIONS: The different types of leukocytes play a key role in the defense of the respiratory tract. The analysis of the distribution of cells in the epithelium, mucosa, and glands of the inferior turbinate confirmed that nonallergic CS is, in fact, chronic, bacterial rhinosinusitis involving the inferior turbinates, and that the pathomechanism is therefore different from that of nasal polyposis.     

Desloratadine activity in concurrent seasonal allergic rhinitis and asthma

Seasonal allergic rhinitis (SAR) and asthma, which are frequently comorbid, share some common allergic pathogenic bases. Clinical manifestations of these disorders might therefore be viewed as local manifestations of a systemic inflammatory state. Not only do the onsets of allergic-rhinitis (AR) and asthma symptoms often coincide (within 1 year), but also nasal challenges with SAR allergens can induce airways hyperreactivity (AHR). Eosinophils, which are key effector cells in both SAR and asthma, cause AHR, tissue damage, and neuronal effects through secretion of toxic granule proteins, enzymes, and other mediators. The novel, nonsedating, histamine H₁-receptor antagonist, desloratadine, which exerts various favorable effects on the allergic cascade, significantly decreased SAR symptoms (e.g., nasal congestion) and diminished daily β₂-agonist use and improved asthma symptoms, while maintaining pulmonary function, in patients with SAR-asthma who were treated with once-daily desloratadine regimens.     

Current status of intranasal glucocorticosteroids in the management of allergic rhinitis

Glucocorticosteroids are the most effective drugs for controlling inflammation of allergic rhinitis (AR). Because of their strong pharmacological action, which can be a so-called 'double-edged sword', glucocorticosteroids are usually taken intranasally so as to reduce their potential for eliciting adverse effects. Accumulating evidence suggests that intranasal glucocorticosteroids control not only nasal symptoms but also ocular symptoms. In contrast to sedating H₁-receptor antagonists, intranasal glucocorticosteroids can improve impaired performance such as daytime sleepiness associated with AR. In Japanese cedar pollinosis, treatment begun immediately after initiation of pollen release or onset of initial symptoms, known as prophylactic (initial) treatment, is recommended. The current version of the practical guideline for management of allergic rhinitis in Japan recommends the use of chemical mediator release inhibitors, second-generation H₁-receptor antagonists, or leukotriene receptor antagonists for prophylactic treatment. However, recent evidence suggests that intranasal glucocorticosteroids might also be useful as first-line drugs for prophylactic treatment. The molecular mechanism of anti-inflammatory action of glucocorticosteroids supports this contention. Moreover, a meta-analysis of studies of intranasal glucocorticosteroids given as monotherapy has revealed that these agents are superior to oral H₁-receptor antagonists and leukotriene antagonists for controlling major symptoms of AR. These findings suggest that glucocorticosteroids, especially intranasal glucocorticosteroids, might be positioned as first-line drugs for the treatment of both perennial and seasonal AR.     

Cyclic AMP-dependent Cl− secretion induced by thromboxane A2 in isolated human colon

Increased release of thromboxane A₂ (TXA₂) has been shown to be involved in inflammatory bowel diseases. In the present study, we have investigated the effect of a stable TXA₂ analogue (STA₂) on the electrical parameters in isolated human colonic mucosa. In the human mucosa set between Ussing chambers, STA₂ stimulated Cl⁻ secretion in a concentration-dependent manner with an EC₅₀ of 0.06 μ m . The STA₂-induced Cl⁻ secretion was significantly inhibited by ONO-3708 (10 μ m ), a specific TXA₂ receptor antagonist. The effect of STA₂ (0.3 μ m ) was independent of the colonic segment from which the tissue was obtained, from caecum to rectum. Chromanol 293B, an inhibitor of the cAMP-dependent KvLQT1 channel, attenuated the STA₂-induced Cl⁻ secretion in the human colonic mucosa (IC₅₀ value 1.18 μ m ). We found that KvLQT1 mRNA and protein were expressed in all the tested segments of the human colon. The STA₂-induced Cl⁻ secretion was significantly inhibited by 8-bromo-2'-monobutyryladenosine-3',5'-cyclic monophosphorothioate (50 μ m ), a membrane-permeant cAMP antagonist. STA₂ (0.3 μ m ) significantly increased the intracellular cAMP levels and the short-circuit current via TXA₂ receptor in a human colonic cell line. These results suggest that the TXA₂-induced Cl⁻ secretion in the colon is mediated via the cAMP pathway in addition to the Ca²⁺–calmodulin pathway which was previously reported.     

Endothelial L-selectin ligand expression in nasal polyps

Background:  L-selectins on leukocytes and their counter-receptors on endothelial cells have been shown to be involved in leukocyte recruitment in chronic rhinosinusitis without nasal polyps (NP).
Objectives:  The purpose of this study was to evaluate the expression level of functionally active endothelial L-selectin ligands in NP obtained from patients with NP of different etiology [simple NP, antro-choanal polyps (ACP) and cystic fibrosis (CF) NP] and inferior turbinate specimens of healthy controls and to compare these levels to the presence of various leukocyte subsets.
Methods:  Nasal polyp specimens and healthy nasal mucosa specimens were obtained from patients undergoing surgery and were immunohistochemically stained with monoclonal antibodies detecting CD34, sialyl Lewis x (sLe^x) of sulfated extended core 1 lactosamines and various leukocyte subsets.
Results:  All NP are characterized by a decrease in the number of CD34+ vessels. The number of eosinophils and the percentage of vessels expressing endothelial sulfated sLe^x epitopes is upregulated in all groups of simple NP. Tissue eosinophilia is increased in those patients with increased disease severity (acetyl salicylic acid intolerance), but the percentage of endothelial sulfated sLe^x epitopes is not. Results on CF NP are similar to those observed for simple NP. Antro-choanal polyps, on the contrary, are characterized by low numbers of tissue eosinophils and relatively few vessels expressing endothelial sulfated sLe^x epitopes.
Conclusions:  Our results suggest that functionally active L-selectin ligands might play a role in guiding leukocyte traffic into NP in patients with simple NP and CF NP but not ACP.     

Glycogen synthase kinase 3 in chronic rhinosinusitis: two faces of a single enzyme in one disease

BackgroundThe origin and pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remain unclear. Glycogen synthase kinase 3 (GSK-3) is a unique multitasking kinase involved in the regulation of inflammation and apoptosis and is an important messenger in the downstream signaling of interleukin 6.ObjectiveTo analyze the possible role of GSK-3 in the pathogenesis of CRSwNP.MethodsWe examined tissue samples of nasal polyps and the inferior turbinate of patients with CRSwNP and the inferior turbinate of individuals without chronic sinusitis (healthy mucosa). Expression levels of GSK-3 and its inactivated form phosphorylated GSK-3 (pGSK-3) were analyzed using DNA microarray, protein array, Western hybridization, and immunohistochemical analysis.ResultsWe found increased expression of GSK-3 in both the nasal polyps and the inferior turbinate of patients with CRSwNP compared with those with healthy mucosa (P < .01). We did not observe a difference between nasal polyps and the inferior turbinate of patients with CRSwNP, but a highly significant increase in the phosphorylation rate of GSK-3 was detected in the tissue of nasal polyps compared with the turbinates of patients with CRSwNP (P < .01).ConclusionGSK-3 may play a crucial role in the inflammatory process in CRSwNP. Nasal polyps originate mainly in the mucosa of the middle meatus of the nose and rarely occur in the region of the inferior turbinate. The inhibition of GSK-3 by phosphorylation in nasal polyps, in contrast to the inferior turbinate, is a possible explanation for the different behavior of the mucosa of the middle meatus and the inferior turbinate.     

Human nasal mucosal carboxypeptidase: Activity, location, and release

Background: Carboxypeptidases (CPs), such as carboxypeptidase N (CPN) (kininase I, E.C.3.4.17.3), may regulate peptide-mediated vasodilation and vascular permeability in respiratory mucosa by degrading proinflammatory peptides such as bradykinin, anaphylatoxins, and neuropeptides during allergic and nonallergic inflammation. The sources of CP activity in human nasal secretions were investigated. Methods: Well-characterized human nasal provocation and secretion analysis methods were used. Potential sources of CPN in human nasal mucosa were identified by immunohistochemistry. CP activity was defined as DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid inhibitable Bz-Gly-Lys degradation. CP activity was measured in nasal mucosal homogenates and nasal lavage fluids induced by methacholine, histamine, and allergen nasal provocation. Results: CPN-immunoreactive material was localized to the glycocalyx of the epithelium, some vessels, and gland ducts near the epithelial basement membrane but not to submucosal gland cells. CP activity in human nasal lavage fluid after saline nasal provocation was 0.10 ± 0.04 U/L. Histamine provoked secretion of significantly more CP activity (3.84 ± 0.99 U/L; p < 0.01 vs saline). Methacholine did not significantly increase secretion (0.54 ± 0.22 U/L). After nasal allergen challenge, CP activity was at a maximum between 11 and 20 minutes, and CP activity correlated with IgG concentration (r = 0.91, p < 0.01), a marker for proteins of plasma origin, suggesting that CP activity originated in plasma. Conclusions: These data suggest that plasma is the predominant source of CP activity secreted from human nasal mucosa and that plasma extravasation and interstitial fluid exudation across the epithelium are the primary processes regulating its appearance in nasal secretions. (J ALLERGY CLIN IMMUNOL 1995;96:924-31.)     

Reduction of soluble ICAM-1 levels in nasal secretion by H1-blockers in seasonal allergic rhinitis

ICAM-1, a transmembrane glycoprotein promoting adhesion in immunologic and inflammatory reactions, was found to be increased on nasal epithelial cells of patients with allergic rhinitis. Loratadinae, an H₁-Mocker, was found to reduce in vitro the expression of ICAM-1 on nasal epithelial cells. A double-blind, parallel-group study was carried out during the pollen season to compare the effect of two H₁-blockers, cetiraizine (10 mg OD) and loratadine (10 mg OD), on the release of soluble ICAM-1 in nasal secretions. A group of untreated patients was used as a control group. sICAM-1 was measured by enzyme immunoassay before and after 2 weeks of treatment. Symptoms were significantly decreased in the actively treated groups. sICAM-1 levels were unchanged in the control group but were significantly reduced in the two treated groups ( P <0.015, Wilcoxon's W test). This study shows that two H₁-blockers, loratadine and cetirizinae, have a similar effect on sICAM-1 released in nasal secretions during the pollen season.