Cells In Nasal Secretions Research Articles

EFFECT OF NASAL CORTICOSTEROIDS ON OXIDATIVE STRESS PARAMETERS IN CHILDREN WITH ALLERGIC RHINITS

Oxidative stress (OS) is a result of an imbalance between the production of free radicals and the antioxidant defense mechanisms in the organism. Allergic rhinitis (AR) is characterized by chronic inflammation and increased presence of eosinophils, basophils and mast cells in nasal secretions and mucosa. Stimulation of these cells results in production of free radicals (reactive oxygen metabolites). The aim of this study was to determine the level of OS and antioxidant capacity in children with AR after 8 weeks treatment with nasal corticosteroid (NCS). Thirty children with AR (mean age 8.3 ± 2.9 years) were included. The FRAS 5 Bravo system was used to determine the OS parameters. Demographic characteristics, medical history, children's living conditions and eating habits were obtained from the questionarrie. Anthropometric measurements, absolute number of eosinophils in the peripheral smear and eosinophils in nasal smear were performed in each child. The mean value of blood concentration of serum peroxides (d-ROMs) was 403.4 ± 91.3 U. Carr, while the mean value of OS index was 95.1 ± 45.9. After 8 weeks of treatment with NCS, the mean values were 337.6 ± 71.9 U. Carr, and 53.6 ± 28.3, respectively. Both parameters had significantly lower values (p<0.05) compared to those before the initiation of NCS therapy. After treatment, a significant improvement in the symptoms score of AR was detected (p<0.05). NCS therapy makes asignificant improvement of the symptoms and a considerable reduction in the OS in children with AR. Keywords: allergic rhinitis, children, oxidative stress, d-ROMs, PAT, OSI

Possible role of highly activated mucosal NK cells against viral respiratory infections in children undergoing haematopoietic stem cell transplantation

Infection is the leading cause of non-relapse-related mortality after allogeneic haematopoietic stem cell transplantation (HSCT). Altered functions of immune cells in nasal secretions may influence post HSCT susceptibility to viral respiratory infections. In this prospective study, we determined T and NK cell numbers together with NK activation status in nasopharyngeal aspirates (NPA) in HSCT recipients and healthy controls using multiparametric flow cytometry. We also determined by polymerase chain reaction (PCR) the presence of 16 respiratory viruses. Samples were collected pre-HSCT, at day 0, +10, +20 and +30 after HSCT. Peripheral blood (PB) was also analyzed to determine T and NK cell numbers. A total of 27 pediatric HSCT recipients were enrolled and 16 of them had at least one viral detection (60%). Rhinovirus was the most frequent pathogen (84% of positive NPAs). NPAs of patients contained fewer T and NK cells compared to healthy controls (p = 0.0132 and p = 0.120, respectively). Viral PCR + patients showed higher NK cell number in their NPAs. The activating receptors repertoire expressed by NK cells was also higher in NPA samples, especially NKp44 and NKp46. Our study supports NK cells relevance for the immune defense against respiratory viruses in HSCT recipients.

Cytological Changes in Nasal Secretions Accompanying Delayed Nasal Response to Allergen Challenge

Background Patients with allergic rhinitis when challenged with allergen develop various types of nasal response, such as an immediate nasal response (INR), late nasal response (LNR), dual late nasal response (DLNR), or delayed nasal response (DYNR), probably because of different hypersensitivity mechanisms. This study was designed to investigate the cytological changes in the nasal secretions (NSs) during the DYNR, beginning 24–32 hours, reaching maximum at 32–36 hours, and resolving within 56 hours after the nasal provocation tests (NPTs) with allergens. Methods In 45 allergic rhinitis patients developing 45 positive DYNRs (p < 0.01), the NPTs and the phosphate-buffered saline (PBS) controls were repeated and supplemented with cytological examination of the NSs and determination of interferon (IFN) γ and IL-4 in nasal lavage fluid (NLF). Results The repeated DYNR (p < 0.001) was accompanied by significant changes (p < 0.05) in the counts of neutrophils, monocytes, lymphocytes, epithelial and goblet cells, and, to a limited extent, of plasma cells and platelets in the NSs and increased concentrations of IFN-γ but not of IL-4, in NLF between 24 and 48 hours after the NPT. No significant cytological changes were found in NSs during the PBS controls (p > 0.1). Conclusion The DYNR is associated with cytological profiles in the NS different from those observed during the INR or LNR. The significant count changes of neutrophils, monocytes, lymphocytes, epithelial and goblet cells in NSs, and increased IFN-γ but not of IL-4 concentrations in the NLF, suggest a possible involvement of the cell-mediated hypersensitivity in the DYNR. These results also emphasize the diagnostic value of NPTs combined with cytological examination of the NSs in patients with nasal allergy.

Editorial.

Patients with allergic rhinitis when challenged with allergen develop various types of nasal response, such as an immediate nasal response (INR), late nasal response (LNR), dual late nasal response (DLNR), or delayed nasal response (DYNR), probably because of different hypersensitivity mechanisms. This study was designed to investigate the cytological changes in the nasal secretions (NSs) during the DYNR, beginning 24-32 hours, reaching maximum at 32-36 hours, and resolving within 56 hours after the nasal provocation tests (NPTs) with allergens.In 45 allergic rhinitis patients developing 45 positive DYNRs (p < 0.01), the NPTs and the phosphate-buffered saline (PBS) controls were repeated and supplemented with cytological examination of the NSs and determination of interferon (IFN) γ and IL-4 in nasal lavage fluid (NLF).The repeated DYNR (p < 0.001) was accompanied by significant changes (p < 0.05) in the counts of neutrophils, monocytes, lymphocytes, epithelial and goblet cells, and, to a limited extent, of plasma cells and platelets in the NSs and increased concentrations of IFN-γ but not of IL-4, in NLF between 24 and 48 hours after the NPT. No significant cytological changes were found in NSs during the PBS controls (p > 0.1).The DYNR is associated with cytological profiles in the NS different from those observed during the INR or LNR. The significant count changes of neutrophils, monocytes, lymphocytes, epithelial and goblet cells in NSs, and increased IFN-γ but not of IL-4 concentrations in the NLF, suggest a possible involvement of the cell-mediated hypersensitivity in the DYNR. These results also emphasize the diagnostic value of NPTs combined with cytological examination of the NSs in patients with nasal allergy.

Nasal lavage natural killer cell function is suppressed in smokers after live attenuated influenza virus

BackgroundModified function of immune cells in nasal secretions may play a role in the enhanced susceptibility to respiratory viruses that is seen in smokers. Innate immune cells in nasal secretions have largely been characterized by cellular differentials using morphologic criteria alone, which have successfully identified neutrophils as a significant cell population within nasal lavage fluid (NLF) cells. However, flow cytometry may be a superior method to fully characterize NLF immune cells. We therefore characterized immune cells in NLF by flow cytometry, determined the effects of live attenuated influenza virus (LAIV) on NLF and peripheral blood immune cells, and compared responses in samples obtained from smokers and nonsmokers.MethodsIn a prospective observational study, we characterized immune cells in NLF of nonsmokers at baseline using flow cytometry and immunohistochemistry. Nonsmokers and smokers were inoculated with LAIV on day 0 and serial nasal lavages were collected on days 1-4 and day 9 post-LAIV. LAIV-induced changes of NLF cells were characterized using flow cytometry. Cell-free NLF was analyzed for immune mediators by bioassay. Peripheral blood natural killer (NK) cells from nonsmokers and smokers at baseline were stimulated in vitro with LAIV followed by flow cytometric and mediator analyses.ResultsCD45(+)CD56(-)CD16(+) neutrophils and CD45(+)CD56(+) NK cells comprised median 4.62% (range 0.33-14.52) and 23.27% (18.29-33.97), respectively, of non-squamous NLF cells in nonsmokers at baseline. LAIV did not induce changes in total NK cell or neutrophil percentages in either nonsmokers or smokers. Following LAIV inoculation, CD16(+) NK cell percentages and granzyme B levels increased in nonsmokers, and these effects were suppressed in smokers. LAIV inoculation enhanced expression of activating receptor NKG2D and chemokine receptor CXCR3 on peripheral blood NK cells from both nonsmokers and smokers in vitro but did not induce changes in CD16(+) NK cells or granzyme B activity in either group.ConclusionsThese data are the first to identify NK cells as a major immune cell type in the NLF cell population and demonstrate that mucosal NK cell cytotoxic function is suppressed in smokers following LAIV. Altered NK cell function in smokers suggests a potential mechanism that may enhance susceptibility to respiratory viruses.

Late phase responses after nasal challenges with allergen and histamine in asthmatic children with perennial nasal allergy

Objective: Late phase response (LPR) is difficult to investigate in patients with perennial nasal allergy because of their continuous presentation with nasal symptoms. Contribution of histamine to the LPR is also controversial. In this study, we investigated whether exogenous histamine can induce LPR in asthmatic patients with perennial nasal allergy to house dust. Methods: A total of 40 asthmatic children were divided into clinical, subclinical and non-rhinitis groups based on their daily nasal symptoms. Changes in nasal patency and in inflammatory cells in nasal secretion were quantitatively measured for 6 h by acoustic rhinometry and light microscopy respectively before and after nasal challenge with allergen or histamine. Results: The allergen challenge produced a significant biphasic decrease in nasal patency in the subclinical group and a marginal decrease in the clinical group, with increases in eosinophils 6 h after the challenge. By contrast, histamine challenge induced significant responses in the clinical group and only a slight response in the subclinical group. Eosinophils also accumulated in nasal secretion of the clinical group to significant levels 6 h after histamine challenge. Eosinophil accumulation following histamine challenge was earlier than that after exposure to allergen. Conclusion: We conclude that LPR can be demonstrated in asthmatic children with perennial nasal allergy. Exposure to exogenous histamine also induced LPR, mediated mainly by eosinophil-related mediators.

Pathogenic mechanisms underlying the clinical symptoms of allergic rhinitis.

This paper reviews our previous studies on an objective evaluation of nasal symptoms, a quantitative determination of biochemical mediators, and inflammatory cells in nasal secretions of atopic patients after nasal allergen challenge (NAC) and during natural allergen exposure. The use of the microsuction technique has proved to be a useful and reliable nasal sampling method permitting quantitative analysis of important mediators in nasal secretions. This has provided accurate data on the activity of some important inflammatory cells such as mast cells, basophils, and eosinophils in allergic rhinitis. Our studies demonstrate that a significant increase in the concentrations of histamine, tryptase, and LTC4 in nasal secretions occurs within seconds or minutes after NAC, and this is accompanied by itching, sneezing, rhinorrhea, and nasal obstruction. The infiltration and activation of eosinophils are found to be the predominant condition during the late-phase reaction (LPR), which is mainly characterized by unilateral and/or bilateral nasal obstruction with little sneezing and rhinorrhea. The latter condition is found to be very much similar to the pathophysiology of patients with ongoing allergic rhinitis. In conclusion, our studies demonstrate that patients with ongoing allergic rhinitis seem to be in a continuous late phase state of eosinophilia and increased mediator release, a condition that can explain priming and nonspecific hyperreactivity of the nasal mucous membrane.

鼻腔開存度へのエメダスチンの効果

The effect of emedastine difumarate after antigen challenge was investigated on nasal passage patency and the number of inflammatory cells in the nasal secretion of allergic rhinitis patients. The results showed that the change in nasal passage patency after 8 weeks of emedastine difumarate administration was significantly inhibited 30 minutes after the antigen challenge. No significant changes in the number of inflammatory cells in nasal secretion was observed during the acute or late phases.

Intranasal challenge with aspirin induces cell influx and activation of eosinophils and mast cells in nasal secretions of ASA‐sensitive patients

Although the mechanism of aspirin-sensitivity seems to be related to inhibition of cyclo-oxygenase by aspirin (ASA), the chain of biochemical events leading to the ASA-induced adverse reaction is not clear, and the contribution of particular mediators and inflammatory cells has not been elucidated. To investigate the involvement of secretory, vascular and cellular mechanisms in the pathophysiology of nasal reactions to aspirin. Six patients with ASA-sensitive asthma/rhinosinusitis and seven ASA-tolerant patients were challenged intranasaly with saline and lysine-acetylsalicylic acid (Lys-ASA) 12 mg, on separate occasions. Nasal lavages were obtained before, and then every 15 min after challenges, and analysed for biochemical and cellular composition. Lys-ASA challenge caused rhinorrhoea, sneezing and nasal congestion with parallel increases in total protein and albumin concentration, albumin % and lysozyme activity in the nasal secretions of ASA-sensitive patients. Concomitant with clinical symptoms, an influx of leucocytes into nasal secretions occurred with significant enrichment in eosinophils (mean prechallenge: 24 +/- 12%, postsaline 27 +/- 9%, postLys-ASA 51 +/- 10%; P < 0.03). The influx of eosinophils into nasal secretions was associated with a remarkable increase in Eosinophil Cationic Protein (ECP) immunoreactivity in five of six patients (mean 9.3 +/- 3.8 micrograms/L and 140.9 +/- 45.8 micrograms/mL before and after Lys-ASA, respectively). At the peak of ASA-induced symptoms an increase in the tryptase level was also observed in five of six patients (mean pre-challenge: 2 +/- 0.1 U/L; postLys-ASA 16 +/- 5 U/L; P < 0.01) suggesting activation of mucosal mast cells. In ASA-tolerant patients Lys-ASA did not induce significant symptoms or changes in the biochemical and cellular composition of nasal secretions. The results show that the ASA-induced nasal adverse reaction involves changes in vascular permeability and serous cell secretion. Both activated eosinophils and mast cells may contribute to the pathophysiology of the ASA-induced reaction in the nasal mucosa.

Correlations between complaints, inflammatory cells and mediator concentrations in nasal secretions after nasal allergen challenge and during natural allergen exposure.

A quantitative determination of the inflammatory mediators was performed and correlated with complaints and the measurement of the inflammatory cells in nasal secretions of 18 seasonal allergic rhinitis patients (group 1) outside the pollen season and 40 symptomatic patients (group 2) with seasonal allergic rhinitis during the pollen season. Ten nonallergic subjects (group 3) were also studied as a normal control group. In group 1, 17 (94%) out of 18 patients had an immediate response of nasal symptoms accompanied by a significant increase of histamine, leukotriene C4 (LTC4), and tryptase 5 min after nasal allergen challenge (NAC). One hour later, a simultaneous increase was seen both in the percentage of the eosinophils and in the eosinophil cationic protein (ECP) concentration. The eosinophil count reached a peak 2 h after NAC with a duration of 8 h, while the highest ECP level was reached only after 24 h with no clear-cut plateau. In group 2, a high percentage of eosinophils was observed. Mostly one observed significantly (p < 0.01) higher concentrations of ECP, LTC4 and histamine but not of tryptase than the baseline values of group 1. The authors concluded that during the pollen season allergic rhinitis reflects mainly a chronic state of allergic inflammation of the nasal mucosa involving various inflammatory components induced by one or more episodes of early-phase type allergic reaction. Infiltration of eosinophils and consequently release of the various late-phase inflammatory mediators into the nasal secretions are certainly believed to be the predominant pathophysiologic condition in the patients.

Quantitative cytology of nasal secretions under various conditions.

Quantitative cytology was performed in nasal secretions of normal control (NC), seasonal allergic rhinitis in season (SAR), perennial allergic rhinitis (PAR), chronic sinusitis with mucoid secretion (MS), and chronic sinusitis with mucopurulent secretion (MPS). The majority of inflammatory cells were neutrophils in NC, MS, and MPS; the majority were eosinophils in SAR and PAR. The concomitant appearance of inflammatory cells in nasal secretions was found, i.e., there were significant correlations between neutrophil and eosinophil counts in MPS, and between eosinophil and basophil counts in SAR. The eosinophil/neutrophil ratio was more than 0.1 in SAR and PAR, but the ratio was less than 0.1 in all NC, all MPS, and in 93% of MS; this indicates that 0.1 in eosinophil/neutrophil ratio is the critical value between allergic and nonallergic nasal diseases.

Indirect evidence of nasal inflammation assessed by titration of inflammatory mediators and enumeration of cells in nasal secretions of patients with chronic rhinitis

Pathophysiologic mechanisms of perennial rhinitis are poorly understood. The characterization of inflammation was studied in nasal lavage of patients with perennial rhinitis by the enumeration of cells involved in the allergic inflammation and the measurement of six mediators released in nasal secretions to determine whether some mediators were relevant for the etiologic diagnosis and the occurrence of symptoms. Ten healthy subjects and 57 patients with perennial rhinitis were placed into four groups according to the symptoms they presented at the time of the study and the origin of the allergy. Allergy was characterized by the history, skin prick tests to standardized allergens, and RAST. Eosinophil protein X (EPX), tryptase, histamine, myeloperoxidase, prostaglandin D 2, and leukotriene C 4/D 4 (LTC 4/D 4) were measured in nasal lavage by enzyme assay or radioimmunoassay. Eosinophils and neutrophils were enumerated after cytocentrifugation of the lavage fluid and May Grunwald Giemsa staining. Tryptase, myeloperoxidase and EPX but not histamine levels were increased in all four patient groups. Eosinophils, LTC 4/D 4, and prostaglandin D 2 were significantly ( p < 0.001, p < 0.03, and p < 0.01) increased in allergic and symptomatic patients. EPX was significantly increased in symptomatic allergic and nonallergic patients. This study suggests the involvement of mast cells, neutrophils, and eosinophils, but the latter cells appear to have a more prominent role. The importance of EPX and LTC 4/D 4 in the characterization of chronic symptomatic rhinitis was also observed.

Intraepithelial Cell Population in the Allergic Nasal Mucosa

Intraepithelial migrating cells are presumed to play an important role in nasal mucosal immunity because these are the first to come into contact with various foreign particles inhaled and deposited on the nasal surface. To elucidate nasal mucosal immunity, we carried out a quantitative study of the overall population of migrating cells and the lymphocyte subsets in the epithelial compartment of nasal mucosa in patients with house-dust nasal allergy, in patients with nonallergic rhinitis (chronic infectious rhinitis) and “normal” persons (as controls), using both light and electron microscopy in combination with an immunoperoxidase technique. Scrapings from the inferior turbinates were collected for examination of the epithelium. In order of predominance, the most numerous migrating cells were lymphocytes, followed by eosinophils, mast cells, neutrophils, and macrophages in the allergy group; in the nonallergy group neutrophils, lymphocytes, followed by macrophages and eosinophils; and in the normal group lymphocytes were the most predominant. Electron microscopic observation revealed globule leukocytes in addition to the other cells. In subjects with nasal allergy, the migrating cell population in nasal secretion differed from scrapings, in that neutrophils and eosinophils were predominant. In our study of the lymphocyte subsets, although CD8+ cells (suppressor/cytotoxic T cells) were the most numerous (followed by CD16+ [natural killer cells], CD4+ [helper/inducer T cells] and CD21+ [B cells] in that order of predominance in the epithelium), no difference was noted in number among CD4+, CD8+, and CD16+ cells in nasal secretions. On the basis of their ultrafine structure, many CD8+ cells and some CD4+ cells had the morphological appearance of activated lymphocytes. Lymphocytes, mast cells, and globule leukocytes, all of which reside in the allergic nasal epithelium, are likely to be involved in the nasal mucosal immunity.

Cytologic changes in the nasal secretions during the immediate nasal response

In 102 randomly selected patients with allergic rhinitis caused by immediate hypersensitivity, nasal provocation tests (NPTs) with allergens were evaluated by means of rhinomanometry, and cytologic examination of the nasal secretions (NSs) was performed. The cells in NS of 117 positive immediate nasal responses (INRs) and in 68 negative INRs, correlating with history and skin tests, and in 102 control challenges with phosphate-buffered saline were stained by modified May-Grünwald-Giemsa, toluidine blue, and Hansel's method. The positive INR was accompanied by significant changes in the count of eosinophils (increase followed by decrease) in 67% of neutrophils (decrease followed by increase) in 40%, goblet cells (increase followed by decrease) in 41%, and basophils (decrease) in 13% of the NSs. No significant changes in the count of other types of cells in the NSs were recorded during most of the cases of INR. No significant changes in the count of individual cell types in NSs were found during most cases of negative INR. During the phosphate-buffered saline control challenges, the individual cell types appeared irregularly, and no significant changes in their count were recorded in any patient. The cytologic examination of NS, evoked by allergen, appears therefore to be a valuable supplementary diagnostic parameter for nasal allergy. The repeated counting of eosinophils in NS, before and after allergen challenge, appears to be the best way to discriminate between positive and negative nasal responses, since the eosinophils demonstrated significant changes in their count during 67% of the positive and only 11% of the negative INRs.

Electron microscope study of basophilic cells in allergic nasal secretions.

In order to elucidate whether basophilic cells in nasal secretion belong to blood basophil or tissue mast cell, basophilic cells in the blood, nasal secretion, and nasal mucous membrane were electron microscopically observed in patients with house dust nasal allergy. The majority of basophilic cells in the nasal secretion was identical with the blood basophil in structure. The blood basophils pass through the vessels and emigrate in the mucous blanket in allergy.

Ultrastructural study of the basophilic granural cells in allergic nasal secretion and mucous membrane (author's transl)

A question has been presented as to whether the basophilic granular cells in allergic nasal secretion belong to the blood basophils or the tissue mast cells. The purpose of this study is to answer this by electron microscopic observation on blood basophils and basophilic granular cells in the nasal secretion and mucous membrane.From the results obtained, two types of basophilic granular cells were observed in both nasal secretion and mucous membrane. One of these, similar to the blood basophil, was small, round or oval in shape and lobulated in the nuleus, with a uniformly fine granular pattern of the specific granule. The other, similar to the tissue mast cell, was large, pleomorphic or oval in shape, mononucleated and varied in the pattern of the specific granule from granule to granule, showing scrolls, lamellae, reticula or granules.The majority of the basophilic granular cells in the nasal secretion belonged to the former type of cells, while the majority in the deeper layer of the mucous membrane belonged to be latter, and two-fifth of cells close to the subepithelial layer belonged to the former and the remaining three-fifth to the latter.Conclusively, the majority of the basophilic granular cells in nasal secretion belongs to the blood basophils, and mainly emigrates from the subepithelial small blood vessels of the nasal mucous membrane through the intraepithelial space.

Basophilic Cells in Allergic Nasal Secretions

There have been several reports on the appearence of basophilic granular cells in respiratory secretion of atopic patients but these cells have not yet been identified. We studied ten nasal allergy patients and seven nonatopic subjects by comparing basophilic granular cell counts in nasal secretion from circulating basophil counts five times a day, 7-8 a. m., at noon, 3 p. m., 6 p. m., and 9-10 p. m.Results indicated that circulating basophil counts were significantly increased at night but decreased in the morning and basophilic granular cell counts in nasal secretion of atopic subjects reversely decreased at night and increased in the morning. In seven nonatopic subjects, the basophil counts were not significantly changed throughout the day and basophilic granular cells in nasal secretion were negative.The present study suggests that basophilic granular cells or part of these cells in nasal allergic patients can be identified as basophils from the peripheral blood.

Basophilic cells in nasal secretions

Basophilic cells in nasal secretions

Respiratory syncytial virus infection of the newborn.

In an outbreak of respiratory syncytial (R.S.) virus infection in a maternity hospital the respiratory illness was of a mild nature and the virus was not found in infants without respiratory symptoms. This confirms the suggestion that R.S. virus can infect infants at a very early age. Rapid diagnosis was achieved by applying the direct fluorescent antibody technique to cells in nasal secretions. This proved to be more sensitive than culture techniques where there was delay between the onset of respiratory symptoms and submission of specimens to the laboratory.

The significance of mast cells in nasal smears from patients with food allergy

Mast cells, as well as eosinophils, must be considered as significant cells in the allergic nasal response. There is a difference of interval between the appearance of the peak of eosinophils and that of mast cells in nasal secretions after allergen provocation. A definite increase of mast cells in nasal secretions suggests food allergy.