Eosinophils, mast cells, and basophils in induced sputum from patients with seasonal allergic rhinitis and perennial asthma: Relationship to methacholine responsiveness

doi:10.1016/S0091-6749(97)70195-7

Journal of Allergy and Clinical Immunology

Volume 100, Issue 1, July 1997, Pages 58-64

https://doi.org/10.1016/S0091-6749(97)70195-7 Get rights and content

Abstract

Objectives: We attempted to determine whether inflammation is present in induced sputum of patients with seasonal allergic rhinitis (AR) as compared with those with perennial asthma (AS) and examined its relationship with bronchial responsiveness to methacholine. Methods: Sputum was induced in 30 patients with seasonal rhinitis in response to grass pollens only and in 15 patients with stable, asymptomatic asthma. The AR group was divided according to methacholine PD₂₀ value: the AR– group (n = 15) had a methacholine PD₂₀ greater than 24 μmol; the AR+ group (n = 15) had a methacholine PD₂₀ ranging between 2.2 and 19.6 μmol. In the AS group, methacholine PD₂₀ ranged between 0.42 and 2.6 μmol. The percentage of eosinophils and metachromatic cells (alcian blue–positive) was assessed in sputum by light microscopy. Tryptase-positive cells and EG2+ cells were identified by immunocytochemistry with the mouse anti-human mast cell–tryptase monoclonal antibody and the monoclonal anti-eosinophil cationic protein antibody. Results: We found that the number of eosinophils in the AS group was greater than that in the AR+ group (p < 0.05) and in the AR– group (p < 0.01). Moreover, the eosinophil count was lower in the AR– group compared with the AR+ group (p < 0.05). Similarly, the number of EG2+ cells was greater in the AS group than in the AR group (p < 0.02) and the AR– group (p < 0.05). Moreover, the EG2+ cell count was lower in the AR– group than in the AR+ group (p < 0.05). The number of mast cells and basophils in the AS group was greater than that in the AR group (p < 0.05 and p < 0.01, respectively). Mast cells in sputum were tryptase-positive. Basophils were present in sputum from 23% of patients with AR and 53% of patients with asthma. There was a significant correlation between methacholine PD₂₀ and eosinophils (p < 0.005) and mast cells (p < 0.02) but not with basophils in those patients showing a measurable methacholine PD₂₀ (AR+ and AS groups). Conclusions: Inflammatory cells are present not only in the airways of patients with asthma but also in airways of patients with seasonal AR, even outside natural exposure. Moreover, we provide evidence for the presence of basophils in sputum of patients with asthma even during clinical remission. The presence of bronchial responsiveness is associated with an increase in the number of eosinophils and metachromatic cells. Our findings are consistent with the hypothesis that eosinophils, as well as mast cells, contribute to bronchial responsiveness not only in AS but also in seasonal AR. (J Allergy Clin Immunol 1997;100:58-64.)

Section snippets

Patients

We selected two groups of consecutive patients who were referred to our outpatient clinic: patients with a history of seasonal AR in response to grass pollen without asthma (AR group) and patients with AS (AS group).

Inclusion criteria for patients with rhinitis were: (1) a history of seasonal rhinitis without cough, wheezing, or shortness of breath during natural exposure, lasting 3 to 5 years; (2) positive skin test responses to grass pollens only; and (3) baseline FEV₁ greater than 80% of

Results

Characteristics of patients with AR and asthma, who successfully accomplished sputum induction, are shown in Table I. We studied 30 patients with AR and 15 patients with asthma. Four patients in each group were former smokers. Seven patients with asthma were using inhaled β₂-agonists as needed. The two groups were not different in regard to gender, age, and baseline FEV₁. Methacholine PD₂₀ values in the AR group were different from those of the AS group (p < 0.001, Table I). Patients in the AR

Discussion

In this study we demonstrated that eosinophils and metachromatic cells are present in induced sputum from patients with seasonal AR even outside exposure to the sensitizing antigen. In these patients, cell counts differed from those of patients with asthma. We found that metachromatic cells in induced sputum had the features of both mast cells and basophils. An important and new observation of this study was that basophils were present in seven of 30 (23%) sputum samples from patients in the AR

References (31)

A Foresi et al.
Inflammatory markers in bronchoalveolar lavage and bronchial biopsy in asthma during remission
Chest
(1990)
L Boulet et al.
Influence of natural antigenic exposure on expiratory flows, methacholine responsiveness, and airway inflammation in mild allergic asthma
J Allergy Clin Immunol
(1993)
S Lam et al.
Differences in mediator release between allergic rhinitis and asthma
J Allergy Clin Immunol
(1991)
M Plaut et al.
Abnormalities of basophil “releasability” in atopic and asthmatic individuals
J Allergy Clin Immunol
(1986)
BL Bradley et al.
Eosinophils, T-lymphocytes, mast cells, neutrophils, and macrophages in bronchial biopsy specimens from atopic subjects with asthma: comparison with biopsy specimens from atopic subjects without asthma and normal control subjects and relationship to bronchial hyperresponsiveness
J Allergy Clin Immunol
(1991)
L Boulet et al.
Influence of natural antigenic exposure on bronchoalveolar lavage in subjects with pollen-induced rhinitis
J Allergy Clin Immunol
(1990)
R Bascom et al.
Basophil influx occurs after nasal antigen challenge: effect of topical corticosteroid treatment
J Allergy Clin Immunol
(1988)
DM Claman et al.
Analysis of induced sputum to examine the effects of prednisone on airway inflammation in asthmatic subjects
J Allergy Clin Immunol
(1994)
FL Jahnsen et al.
Erroneous immunohistochemical application of monoclonal antibody EG2 to detect cellular activation
Lancet
(1994)
R Djukanovic et al.
Mucosal inflammation in asthma
Am Rev Respir Dis
(1990)

R Beasley et al.

Cellular events in the bronchi in mild asthma and after bronchial provocation

Am Rev Respir Dis

(1989)

WJ Stevens et al.

Bronchial hyperreactivity in rhinitis

Eur J Respir Dis

(1983)

R Djukanovic et al.

Bronchial mucosal manifestations of atopy: a comparison of markers of inflammation between atopic asthmatics, atopic non asthmatics and healthy controls

Eur Respir J

(1992)

PG Gibson et al.

Cellular characteristics of sputum from patients with asthma and chronic bronchitis

Thorax

(1989)

JV Fahy et al.

Cellular and biochemical analysis of induced sputum from asthmatic and from healthy subjects

Am Rev Respir Dis

(1993)

Cited by (176)

Tissue mast cell counts may be associated with decreased severity of gastrointestinal acute GVHD and nonrelapse mortality
2020, Blood Advances
The functions of mast cells in human graft-versus-host disease (GVHD) are unknown. We studied 56 patients who had an allogeneic hematopoietic cell transplantation (alloHCT) with a biopsy for diagnosis of gastrointestinal tract (GIT) GVHD before any treatment (including steroids): 35 with GIT GVHD, 21 HCT recipients whose biopsies did not confirm GVHD, and 9 with a new diagnosis of inflammatory bowel disease (IBD) as a comparison. The median number of mast cells (mean of CD117⁺ cells, counted in 3 selected spots under 40× magnification) was similar between patients with GVHD (59 cells) and those without GVHD (60 cells). However, the median number of mast cells was significantly associated with maximum clinical stage of GIT GVHD; the lowest counts of mast cells were observed in the highest clinical stage of GIT GVHD (stage 1, 80; stage 2, 69; stage 3, 54; stage 4, 26; P = .01). Moreover, every decrease by 10 mast cells was associated with increased nonrelapse mortality through 1 year (hazard ratio, 0.77; 95% confidence interval, 0.59-1.00; P = .05). AlloHCT recipients all had significantly fewer mast cells, even those without GVHD compared with those with IBD (median, 59 vs 119; P < .01). The median number of GIT mast cells was also significantly lower in patients who received myeloablative conditioning (61.5 cells) than in those who received reduced intensity conditioning (78 cells) in the entire study population (P = .02). We conclude that GIT mast cells are depleted in all alloHCT patients, more prominently in those receiving myeloablative conditioning and those with severe GIT GVHD. Our novel findings warrant further investigation into the biological effects of mast cells in GIT GVHD.
Relationship between various cytokines implicated in asthma
2019, Human Immunology
Citation Excerpt :
A clinical study involving steroid-resistant and steroid-sensitive asthmatics found that BAL cells from steroid-resistant asthmatics expressed higher levels of IL-4 mRNA compared with steroid-sensitive asthmatics [73]. Interleukin 3 is a pluripotential haematopoietic growth factor that acts together with other cytokines such as GM-CSF to promote the development of erythroid progenitors [75], megakaryocytes [75], neutrophils [74], eosinophils [20], basophils [20], mast cells [20] and monocytes [75]. It is generally produced by activated Th cells [136] and mast cells [75].
Asthma is a complex disorder involving immunologic, environmental, genetic and other factors. Today, asthma is the most common disease encountered in clinical medicine in both children and adults worldwide. Asthma is characterized by increased responsiveness of the tracheobronchial tree resulting in chronic swelling and inflammation of the airways recognized to be controlled by the T-helper 2 (Th2) lymphocytes, which secrete cytokines to increase the production of IgE by B cells. There are many cytokines implicated in the development of the chronic inflammatory processes that are often observed in asthma. Ultimately, these cytokines cause the release of mediators such as histamine and leukotrienes (LT), which in turn promote airway remodeling, bronchial hyperresponsiveness and bronchoconstriction. The CD4⁺ T-lymphocytes from the airways of asthmatics express a panel of cytokines that represent the Th2 cells. The knowledge derived from numerous experimental and clinical studies have allowed physicians and scientists to understand the normal functions of these cytokines and their roles in the pathogenesis of asthma. The main focus of this review is to accentuate the relationship between various cytokines implicated in human asthma. However, some key findings from animal models will be highlighted to support the discoveries from clinical studies.
Bronchoprovocation Testing in Asthma: An Update
2018, Immunology and Allergy Clinics of North America
Citation Excerpt :
Among nonatopics, elevated allergen-specific and polyclonal IL-10 production, TNFα, and IFNγ were associated with BHR, whereas BHR among atopic individuals was associated with eosinophilia, IL-5, and IgE.64 There are numerous epidemiologic and clinical trial studies that present associations, albeit modest, between BHR and features of atopy or other biomarkers of eosinophilic inflammation in the airway (from sputum, bronchoalveolar lavage, and exhaled substances).61,65–73 The associations with various measures of atopy can be variable.
Evaluation of vascular endothelial growth factor-A and Endostatin levels in induced sputum and relationship to bronchial hyperreactivity in patients with persistent allergic rhinitis monosensitized to house dust
2015, Revista Portuguesa de Pneumologia (English Edition)
Citation Excerpt :
Although the inflammatory pathogenesis of BHR is understood in asthmatic patients, light has not been shed on the precise mechanism of BHR in patients with AR.11,19,20 In our trial, we demonstrated higher eosinophil numbers in patients with PAR compared to healthy controls independent of BHR, as in previous studies including our first study in allergic rhinitis patients monosensitized to pollen.10,21–25 Also, the percentage of eosinophils in induced sputum was found to be significantly higher in PAR patients with BHR when compared with those without BHR The presence of sputum eosinophils in our patients seemed to be linked to the presence of both AR and BHR.
Studies about the pathogenesis of bronchial hyperreactivity (BHR) in patients with persistent allergic rhinitis (PAR) and its relationship with lower airway remodeling are extremely limited.
This study evaluated bronchial vascular remodeling via the measurement of angiogenic factor, vascular endothelial growth factor-A (VEGF-A), and anti-angiogenic factor, Endostatin, and evaluated their relationship with BHR in patients with PAR.
The study group consisted of 30 patients with PAR monosensitized to house dust mites and 14 non-allergic healthy controls. All subjects underwent induced sputum and methacholine (M) bronchial provocation tests. VEGF-A and Endostatin levels were measured by ELISA in induced sputum supernatants.
The percentages of eosinophils in induced sputum were significantly increased in patients with PAR compared with healthy controls. There were no significant differences between patients with PAR and healthy controls in terms of levels of VEGF (37.9 pg/ml, min–max: 5–373 pg/ml vs. 24.9, min–max: 8–67 pg/ml, p = 0.8 respectively), Endostatin (532.5 pg/ml, min–max: 150–2125 pg/ml vs. 644, min–max: 223–1123 pg/ml, p = 0.2 respectively) and VEGF/Endostatin ratio (0.057 vs. 0.045, p = 0.8 respectively). In addition, there were no significant differences between patients who are BHR positive (n = 8), or negative to M (n = 22) in terms of levels of VEGF, Endostatin and VEGF/Endostatin ratio and no correlations among value of PD20 to M and levels of VEGF, Endostatin and VEGF/Endostatin ratio.
We conclude that VEGF-A and Endostatin did not differ between patients with PAR and healthy controls regardless of BHR to M.
Bronchial inflammation in seasonal allergic rhinitis with or without asthma in relation to natural exposure to pollen allergens
2015, Allergologia et Immunopathologia
Nasal inflammation in allergic rhinitis enhances bronchial Th2 driven inflammation and development of asthma. We assessed bronchial inflammation induced by natural allergen exposure during pollen season in patients with pollinosis with or without asthma to show the intensity of inflammation in asthma and rhinitis and possible persistence of inflammation in periods without allergen exposure.
Sputum was induced in 52 patients with seasonal allergic rhinitis without asthma, 38 patients with seasonal allergic rhinitis and seasonal asthma and 23 healthy volunteers. Sampling was performed 6–8 weeks before the expected beginning of symptoms, during symptomatic period and 6–8 weeks after the end of symptoms. Sputum ECP was measured by means of chemiluminiscent immunometric assay and sputum cell counts were assessed by classical staining and immunocytochemistry.
Sputum eosinophils were on the whole higher in both asthma and rhinitis compared to controls (p < 0.001, p = 0.003). The rise of eosinophils during pollen season compared with values out of pollen season was significant in asthma (classical staining) (p = 0.014) and slightly apparent in rhinitis (immunocytochemistry) (p = 0.073). The seasonal rise of sputum ECP was observed only in rhinitis (p = 0.006).
Inflammation of the lower airway in patients with allergic rhinitis with and without asthma has been confirmed by means of both sputum eosinophil count and sputum ECP level. Persistent inflammation of lower airway in periods without allergen exposure was proven in seasonal asthma. This may have implications for the therapy of seasonal allergic rhinitis with and without asthma in terms of promoting long-term anti-inflammatory treatment.
Bronchial hyperresponsiveness in seasonal allergic rhinitis patients is associated with increased IL-18 during natural pollen exposure
2012, Cytokine
The mechanism of bronchial hyperresponsiveness (BHR) is not certain in seasonal allergic rhinitis (SAR) patients.
We aimed to investigate the effects of natural pollen exposure on IL-18 and its relationship with BHR.
Thirty-two SAR patients with grass pollen sensitivity, 14 nonallergic rhinitis (NAR) patients and 17 normal-controls were included. Sixteen SAR patients had BHR during pollen season and off-season. Serum IL-18 levels were measured in SAR patients during pollen season between May–August and off-season between November–February. IL-18 levels were measured in NAR patients and normal controls once.
During pollen season, SAR patients with BHR had significantly increased levels of IL-18 than those without BHR (279.2 ± 161.1 versus 145.3 ± 101.0 pg/ml, p = 0.012). Serum IL-18 levels were not different between SAR patients with and without BHR during off-season (233.8 ± 139.7 versus 183.2 ± 162.9 pg/ml, p = 0.16). Serum IL-18 levels in SAR patients during pollen season (212.3 ± 148.8 pg/ml) and off-season (208.5 ± 151.5 pg/ml) were not different than those NAR patients (224.8 ± 180.1 pg/ml, p = 0.98 and p = 1.0, respectively) and normal controls (174.8 ± 76.0 pg/ml, p = 0.60 and p = 0.76, respectively).
The results suggested us that BHR in SAR patients is associated with increased IL-18 during natural pollen exposure.

View all citing articles on Scopus

^☆: From Servizio di Fisiopatologia Respiratoria, Modulo di Allergologia ed Immunopatologia Polmonare, Sesto San Giovanni, and Istituto di Clinica delle Malattie dell'Apparato Respiratorio, Università di Parma.

^☆☆: Reprint requests: Antonio Foresi, MD, Servizio di Fisiopatologia Respiratoria, Viale Matteotti 83, 20099 Sesto San Giovanni, Italy.

^★: 1/1/81444

View full text

Eosinophils, mast cells, and basophils in induced sputum from patients with seasonal allergic rhinitis and perennial asthma: Relationship to methacholine responsiveness☆,☆☆,★

Abstract

Section snippets

Patients

Results

Discussion

Chest

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

Lancet

Mucosal inflammation in asthma

Am Rev Respir Dis

Cellular events in the bronchi in mild asthma and after bronchial provocation

Am Rev Respir Dis

Bronchial hyperreactivity in rhinitis

Eur J Respir Dis

Bronchial mucosal manifestations of atopy: a comparison of markers of inflammation between atopic asthmatics, atopic non asthmatics and healthy controls

Eur Respir J

Cellular characteristics of sputum from patients with asthma and chronic bronchitis

Thorax

Cellular and biochemical analysis of induced sputum from asthmatic and from healthy subjects

Am Rev Respir Dis