Suppression of the late asthmatic reaction by hyposensitization in asthmatic children allergic to house dust mite (Dermatophagoides pteronyssinus)

In this study, we wanted to test the hypothesis that hyposensitization (HS) abrogates the late asthmatic reaction (LAR). We therefore selected 15 asthmatic children (subjects) sensitized to house dust mite (HDM), as proven by positive prick tests and/or specific IgE via the RAST. All children demonstrated a positive bronchial provocation test (BPT) to HDM; 14 showed a dual asthmatic reaction and one child showed an isolated LAR. All children were treated with anti-asthmatic drugs and received HS. They were rechallenged after 1 yr of HS while their anti-asthmatic medication was not changed, except for beta-agonists which were used only for relief of symptoms. Two BPTs, with a mean interval of 6.4 months, were also performed in eight asthmatic children (controls), who did not receive HS. In the controls, the same reaction pattern was observed during both BPTs. In the subjects, the LAR completely resolved in 5/15 after 1 yr of HS (P = 0.04). Furthermore, as a group, the subjects showed a less severe LAR after 1 yr of HS (expressed as mean fall of FEV1) (P less than 0.0001). The PD20 of the immediate asthmatic reaction (IAR) was the same as before HS was started, but the IAR was also less severe after 1 yr of HS (expressed as mean fall of FEV1) (P = 0.028). From these observations it is concluded that, in asthmatic children, HS may resolve and/or reduce the severity of the LAR. Although the PD20 of the IAR is not changed, the severity of the IAR is also reduced after 1 yr of HS.     

Effect of a bronchial provocation test with house-dust mite on blood eosinophilia, eosinophil cationic protein, soluble interleukin-2 receptor, and interleukin-6 in asthmatic children

Eighteen children with perennial asthma and allergy to house-dust mite (HDM) underwent a bronchial challenge with HDM. Before and 24 h after the test, a venous blood sample was taken to determine levels of eosinophils, eosinophil cationic protein (ECP), soluble interleukin-2 receptor (IL-2R), and interleukin-6 (IL-6). A histamine challenge was performed before and 24 h after the HDM challenge. All subjects showed an immediate asthmatic reaction (IAR). A definite late asthmatic reaction (LAR) was observed in 15 children, a probable LAR in two, and no LAR in one. Because of persistent bronchial obstruction (FEV₁>70%), eight children were unable to perform a histamine challenge 24 h after the allergen challenge. These were the children with the lowest prechallenge provocation dose (PD₂₀) of histamine. In the other 10 children, the mean PD₂₀ histamine decreased after the HDM challenge (mean PD₂₀ before was 0.56 mg/ml; after challenge it was 0.14 mg/ml; P= 0.007). After the HDM challenge, an increase was detected in the mean values of blood eosinophils (mean before was 446/mm³; mean after was 733/mm³; P= 0.002), ECP (mean before was 26.3 μg/1; mean after was 34.3 μg/1; P<0.040), and IL-2R (mean before was 116.35 U/ml; mean after was 128.52 U/ml; P<0.040). On the other hand, IL-6 remained unchanged after the HDM challenge (mean before was 9.47 pg/1; mean after was 9.70 pg/1; P= 0.360). Furthermore, as compared with a group of normal, age-matched children (n =18), asthmatic children were found to have higher prechallenge levels of ECP (mean: 10.3 μg/1 compared with 26.3 μg/1) (P>0.001) and IL-2R (mean: 80.30 U/ml compared with 116.35 U/ml) (P =0.009), but not of IL-6 (mean: 11.34 pg/1 compared with 9.47 pg/1) (P = 0.436). A correlation was found between the duration of asthma and the severity of the LAR expressed as area under the curve (AUCLAR) (r = 0.50; P<0.040). Furthermore, a correlation was detected between the level of total IgE and the level of ECP (r = 0.51; P<0.030). The decrease in FEV₁ during the LAR tended to correlate with the increase of IL-2R (r = 0.48; P = 0.050). This tendency was not found with the increase of eosinophils, nor with the increase of ECP. We conclude that both lymphocytes and eosinophils are activated by an allergen challenge, but that only the activation of lymphocytes tends to correlate with the LAR, suggesting that lymphocytes are also closely involved in the pathogenesis of the allergen-induced LAR.     

Evolution of the late asthmatic reaction during immunotherapy and after stopping immunotherapy

In previous studies it was demonstrated that the frequency and the severity of the late asthmatic reaction (LAR) can be attenuated by immunotherapy (IT). The present study was set up to observe the evolution of the LAR under IT and after having stopped IT. Nineteen children with bronchial asthma and an LAR to house dust mite (Dermatophagoides pteronyssinus) (HDM) were selected for this study. All subjects received IT with HDM extract during 1 year. Thereafter, the children were divided randomly into two groups to receive, double-blind, during a second year, IT with HDM (N = 9) or placebo injections (N = 10). Bronchial challenges were performed after the first and second year. After the first year, a significant decrease in the severity of the LAR was noted in all but one subject (mean decrease of FEV1 before, 40.53% versus after 1 year, 22.95%; p less than 0.0001). After the second year, the severity of the LAR remained the same in the group that received HDM injections during the second year (20.78% versus 23.00%), but in the group that received placebo injections during 1 year, a significant worsening of the LAR was observed after the second year (24.90% versus 31.20%; p = 0.038). From this study it can be concluded that the severity of the LAR decreases after the first year of IT but that the severity of the LAR remains the same after the second year of IT. In children who stop receiving IT after 1 year, we observed a recurrence of the LAR after 1 year to the same level as before IT was started.     

Frequency and intensity of late asthmatic reactions after bronchial allergen challenge in asthma and rhinitis

The occurrence of late asthmatic reactions after bronchial allergen challenge was studied in 50 house dust mite allergic patients subdivided in three groups: one group had asthma without nasal symptoms, another group had rhinitis without pulmonary symptoms and a third group had a combination of both asthma and rhinitis. Late asthmatic reactions were present in 80% of asthmatic patients and in 18.7% of rhinitis patients. The degree of non-specific bronchial reactivity to histamine (provocative dose 15 or PD15 histamine) and the degree of immediate reactivity to allergen (PD15 house dust mite) did not differ significantly between patients with and without late asthmatic reactions. These findings suggest that an important difference between asthma and rhinitis is the lack of late asthmatic reactions in rhinitis patients, whereas the degree of immediate bronchial reactivity to the allergen is similar in asthma and rhinitis.     

Lymphocyte transformation test with house dust mite (Dermatophagoides pteronyssinus) in normal children, asthmatic children and asthmatic children receiving hyposensitization

In the first part of this study the proliferative response of lymphocytes (lymphocyte transformation test) to house dust mite (HDM) stimulation in cultures was studied in normal children (n= 16), asthmatic children who never received hyposensitization (HS) (n = 50) and asthmatic children receiving HS with HDM for at least 6 months (n = 20). The results are expressed as disintegrations per minute (d.p.m.) and as stimulation index (SI = d.p.m. in the presence of the allergen/d.p.m. in the control culture). A positive SI (> 2) was found in 54% of the asthmatic children who never received HS, in 30% of the asthmatics receiving HS and in none of the normal children. Furthermore, between asthmatics with and without HS, the SI was not statistically different, although asthmatics without HS tended to have a higher SI (median value: 2.13 vs 1.38) (P= 0.10). In a second series of experiments the effect of adding interleukin-2 (IL-2) to the lymphocyte cell culture was studied in asthmatic children with and without HS. Interleukin-2 induced an additional stimulatory effect on the lymphoproliferative response to HDM and to phytohaemagglutinin in patients who never received HS, but had no effect in patients receiving HS. We conclude that HS treatment seems to have an inhibiting effect upon this proliferative response, not only inhibiting the degree of the allergen-induced lymphocyte proliferation, but also inhibiting the sensitivity of proliferating lymphocytes for IL-2. These inhibiting effects upon lymphocytic activation could be responsible for the anti-inflammatory effects (i.e. suppression of the late asthmatic reaction) of HS.     

Changes in serum haptoglobin level after allergen challenge test in asthmatic children

Bronchial asthma is characterized by airway inflammation, which underlies the phenomenon of bronchial hyperresponsiveness. Previous studies have shown that this correlates with the serum concentration of haptoglobin. The occurrence of the late asthmatic response (LAR) after an allergen challenge test is associated with airway inflammation. The objectives of this study were to examine serum levels of haptoglobin during the 24 h after allergen challenge and to compare changes between the subjects with and without LAR. We studied two groups of children with perennial asthma who developed the early asthmatic response (EAR) only (group 1: n = 14), and EAR but also LAR (group II: n = 14) after an allergen (Dermatophagoides pteronyssinus) challenge test. Serum concentrations of haptoglobin were measured at baseline, at EAR, and at 2 h (recovery), 8 h (LAR), and 24 h after the challenge. Baseline levels were similar in the two groups (group I: 128±57 mg/dl: group II: 129±50 mg/dl). In group I, there was no significant change in the level at any time point; in contrast, the subjects in group II showed a relative fall (92±40 mg/dl) at 8 h, and an increase (161±79 mg/dl) at 24 h after the challenge. Our results indicate that the serum concentration of haptoglobin decreases at the time of LAR and is subsequently replenished during the ensuing time. Although further studies are needed, we think that haptoglobin may be inflused into the airways during the inflammatory process associated with LAR, and that this may be followed by "overshooting" production.     

The late asthmatic response is associated with baseline allergen-specific proliferative responsiveness of peripheral T lymphocytes in vitro and serum interleukin-5

BACKGROUND: Increasing insights into the mechanism underlying the allergen-induced late asthmatic response (LAR) have been gained with implication of activated eosinophils and CD4+ T lymphocytes. However, the patient characteristics that indicate the individual capacity to develop a LAR are not well-defined. METHODS: In 22 subjects with mild to moderate house dust mite-allergic asthma, we investigated the relationship between the LAR and two other models of late-phase allergic inflammation, i.e. the allergen-specific proliferative response of peripheral blood T lymphocytes in vitro and the late cutaneous response. Non-specific bronchial responsiveness (PC20histamine), lung function (FEV1), peripheral blood eosinophil count, early phase allergic skin sensitivity, and levels of total and specific immunoglobulin E (IgE) were determined prior to bronchial allergen challenge. Serum levels of interleukin-5 (IL-5) were measured before and at several time points after allergen inhalation. RESULTS: A significant correlation was found between the magnitude of the LAR and the allergen-specific proliferative response of peripheral T lymphocytes (r = 0.44, P = 0.04) but not the late cutaneous response. Stepwise-multiple linear regression of the magnitude of the LAR on the parameters analysed at baseline, resulted in a model combining PC20 histamine, early phase allergic skin sensitivity, and the allergen-specific proliferative response of peripheral T lymphocytes (R2 = 0.84, P<0.001). No contribution of the late cutaneous response to the prediction of the LAR was found. Serum levels of IL-5 increased significantly at 6 h (P = 0.01) and 24 h (P = 0.003) after bronchial allergen challenge and correlated with the allergen-specific proliferative response of peripheral T lymphocytes in vitro (rho = 0.48, P = 0.02). CONCLUSIONS: The findings in this study point to a role of TH2-lymphocyte responses in the development of the allergen-induced LAR. In allergic asthmatic patients, allergen-specific responsiveness of peripheral T-lymphocytes in vitro may serve as a model to determine the individual capacity to develop a LAR after allergen inhalation.     

Vascular permeability and airway narrowing during late asthmatic response in dogs treated with Metopirone.

Recently, we have developed an animal model of late asthmatic response (LAR) by treating naturally sensitized dogs to Ascaris suum antigen with the cortisol-synthesizing inhibitor, Metopirone. By using this animal model, we examined the contribution of edema in the airway wall to the development of LAR. To study whether airway microvascular leakage is increased in association with LAR, we performed antigen challenge in dogs treated with Metopirone. We measured the amount of extravasated Evans blue (EB) dye from the esophagus, trachea, and large and small bronchi 8 hours after the antigen challenge in dogs demonstrating immediate asthmatic response alone (IAR) and in dogs demonstrating both IAR and LAR. Airway responses to A. suum antigen were assessed by changes in respiratory resistance measured with the force oscillation technique at 3 Hz. EB dye extravasation did not increase significantly from that of control in any tissues in IAR (P greater than 0.10), but in LAR, it increased significantly from that of control (p less than 0.01) and IAR (p less than 0.05) in large and small bronchi. Histologic assessment of vascular permeability revealed that Monastral blue-labeled leaking vessels were only in sections from LAR, and leaking vessels were limited to small vessels (10 to 25 microns) in the trachea, large (diameter, greater than 5 mm) and small bronchi (2 to 4 mm in diameter), and bronchiole. The permeability index defined as the ratio of area of small vessels labeled with Monastral blue to that of the total small vessels in the walls was highest in the small bronchi. LAR significantly increased submucosal thickness of the small bronchi (p less than 0.05) compared with that in IAR. Both EB dye extravasation and permeability index in large and small bronchi also significantly increased during IAR within 3 minutes after the antigen challenge (p less than 0.05), but IAR did not alter the submucosal thickness of the small bronchi. These results imply that the increase in vascular permeability and submucosal thickness, especially in small bronchi, may be an important factor in the pathogenesis of LAR.     

The changes in plasma beta-thromboglobulin (beta-TG), platelet factor 4 (PF4) and thromboxane B2 (TXB2) after a bronchial provocation test (BPT) with house dust (HD) allergen]

In order to investigate the role of platelets in allergic asthma, the time related changes in plasma levels of beta-TG, PF4 and TXB2 were evaluated following BPT with HD in 19 patients with bronchial asthma who were positive in skin test and RAST to HD. The results obtained were as follows. 1) Plasma beta-TG and PF4 levels tended to increase following BPT with HD at the time of immediate asthmatic response (IAR) in patients showing IAR alone. 2) Plasma beta-TG and PF4 levels increased significantly (p less than 0.05) at IAR and tended to increase at the time of late asthmatic response (LAR) in patients showing a dual asthmatic response (DAR). 3) The levels of plasma TXB2 in patients showing IAR alone significantly increased at IAR (p less than 0.05) and gradually decreased and the levels of plasma TXB2 in patients showing a DAR increased in each period of IAR, 3 hr after BPT and LAR, and the peak of TXB2 was observed in 3 hours after BPT. 4) These results suggest that platelets are activated at IAR and there was also a possible activation in platelets at LAR.     

Effect of budesonide and montelukast in asthmatic children exposed to relevant allergens

BACKGROUND: Montelukast has been shown to be effective in controlling the increase in exhaled NO in asthmatic children re-exposed to house dust mite (HDM). This study compared the effect of low dose inhaled budesonide and oral montelukast in preventing the expected relapse of airway inflammation and reactivity in a group of 24 mild asthmatic children allergic to HDM after a brief period of exposure to relevant allergens. METHODS: Lung function, bronchial hyperresponsiveness (BHR) to methacholine (PC(20)), fractional exhaled nitric oxide (FeNO) levels and sputum eosinophilia were evaluated. RESULTS: Pulmonary function remained stable. The BHR was unchanged after exposure in the group treated with budesonide, whereas a significant increase (P = 0.028) was observed in the patients receiving montelukast. No significant difference was observed in FeNO levels after exposure to mite antigen in the two groups. In both the groups of asthmatic children we observed a significant increase in sputum eosinophil % after the exposure to mite antigen. CONCLUSIONS: The significant increase in BHR level observed in the group of children receiving montelukast suggests a more comprehensive effect as disease controller by inhaled steroids than by leukotriene antagonist in allergic asthmatic children re-exposed to relevant allergens.     

Magnitude of late asthmatic response to allergen in relation to baseline and allergen-induced sputum eosinophilia in mild asthmatic patients.

BACKGROUND: Late asthmatic response (LAR) to allergen challenge is a validated method for studying the pathogenesis of and new treatments for asthma in the laboratory. OBJECTIVE: To evaluate the relationship between the magnitude of allergen-induced LAR and clinical and biological determinants, including sputum and blood eosinophil percentages and eosinophil cationic protein concentrations. METHODS: Thirty-eight untreated mild asthmatic patients (mean age, 21.2 years) were selected for the presence of allergen-induced early asthmatic response (EAR) and LAR. Each patient measured methacholine responsiveness (provocation dose that caused a decrease in forced expiratory volume in 1 second of 20% [PD20FEV1]) at baseline, differential blood cell counts and eosinophil cationic protein levels in blood and induced sputum, and serum neutrophil chemotactic activity at baseline and 24 hours after allergen challenge. RESULTS: A correlation was found between LAR (as area under the curve [AUC]) and sputum eosinophil percentages at baseline (r = 0.51; P = .001) and 24 hours after allergen challenge (r = 0.44; P < .007). Furthermore, we found significant correlations between AUC LAR and AUC EAR, baseline methacholine PD20FEV1, baseline blood eosinophil percentages, and baseline serum neutrophil chemotactic activity. A stepwise multiple regression analysis showed that the stronger determinants of AUC LAR were baseline sputum eosinophilia and AUC EAR. CONCLUSION: Baseline sputum eosinophilia and functional findings are determinants of the magnitude of allergen-induced LAR.     

Effect of ciclesonide on allergen challenge in subjects with bronchial asthma

BACKGROUND: The aim of this clinical trial was to investigate whether repeated inhalation of the new inhaled steroid ciclesonide reduces the early-phase (EAR) and late-phase (LAR) reactions after allergen challenge in patients with mild allergic asthma. Also, this study provides further data on safety and tolerance of ciclesonide. METHODS: The study was designed as a double-blind placebo-controlled randomized crossover trial. Following a baseline period, patients were randomized to either of two treatment sequences (ciclesonide/placebo, placebo/ciclesonide) each of which lasted for one week and were separated by 3-5 weeks from the alternate treatment sequence. Patients received 800 micro g ciclesonide twice daily by means of a Cyclohaler. At the end of each treatment patients were subjected to an allergen challenge. RESULTS: Thirteen asthmatic patients (mean FEV1 of 91% predicted) who experienced an EAR and LAR after allergen challenge participated in the study. The time-average FEV1 decreases 0-2 h (2-12 h) after allergen challenge as measure of the EAR (LAR) were significantly reduced (P < 0.05, one-sided) from 0.426 L to 0.233 L (EAR) and from 0.443 L to 0.213 L (LAR), respectively. Thus, the study results suggest that ciclesonide significantly lowered the extent of EAR and LAR compared to placebo. Ciclesonide was well tolerated and no drug-related adverse events were reported. Cortisol excretion in 24-h urine showed no significant difference between ciclesonide and placebo. CONCLUSIONS: The study supports the efficacy and safety of ciclesonide.     

Inflammatory cells and eicosanoid mediators in subjects with late asthmatic responses and increases in airway responsiveness

To determine the relationship of inflammatory cells and eicosanoid mediators to the pathogenesis of the late asthmatic response (LAR) and increases in nonspecific airway responsiveness, we studied bronchoalveolar lavage (BAL) cells and fluid in 27 subjects 12 hours after inhaled antigen challenge. Methacholine challenge was performed before antigen challenge and 24 hours later (12 hours after BAL). Eight subjects had no LAR (-LAR, less than or equal to 10% fall in FEV1), nine subjects had an equivocal LAR (+/- LAR, 11% 25% fall in FEV1), and 10 subjects had a definite LAR (+LAR, greater than 25% fall in FEV1). Subjects developing +LAR had increased airway responsiveness at baseline compared with that of subjects developing an +/- LAR, but not with subjects having -LAR. If airway responsiveness was markedly increased at baseline, further increases after antigen challenge were often not observed. We found that both percent neutrophils and eosinophils increased in BAL as the severity of the LAR increased, but significant differences between the groups with -LAR and +LAR were only observed when both cell types were considered together. In addition, there was a significant correlation between the combined cell percentages and the severity of the LAR as determined by fall in FEV1. Likewise, increases in airway responsiveness were associated with significant increases in both neutrophil and eosinophil numbers, but only neutrophils correlated with the change in airway responsiveness after antigen challenge. However, despite the significant physiologic and cellular differences that we found between our groups, no significant differences could be found in BAL eicosanoid-mediator concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Der p 1 and Der p 2 induce less severe late asthmatic responses than native Dermatophagoides pteronyssinus extract after a similar early asthmatic response

Background The models for exposure to house dust in research and clinical practice are selected with respect to their role in IgE‐mediated immediate hypersensitivity. The use of isolated major allergens instead of complex allergen extracts is becoming increasingly popular as it offers some important advantages for quantitative measures in diagnosis and research. Objective To compare house dust mite extract and isolated mite major allergens with respect to their ability to induce early and late asthmatic responses and bronchial hyperreactivity. Methods Bronchial responses to house dust mite (HDM, Dermatophagoides pteronyssinus) extract and isolated major allergens from HDM (Der p 1 and Der p 2) were compared in a double‐blind, randomized, cross‐over study in 20 patients with mild to moderate asthma who were allergic to HDM. Allergen was titrated to a standardized early asthmatic response. Bronchial hyper‐responsiveness to histamine (PC₂₀histamine) was determined before and after allergen inhalation to assess allergen‐induced bronchial hyper‐responsiveness and IL‐5 was measured in serum. In addition, the allergens were applied in intracutaneous skin tests and activation of basophil leucocytes and proliferation of peripheral blood mononuclear cells was tested in vitro. Results After a similar early asthmatic response (mean Δforced expiratory volume in 1 s (FEV₁)_,max?29.4 (SD 7.2) vs. ?33.1 (8.6) %; mean difference 3.6 (95% CI ?0.9 to 8.2) %), the late asthmatic response (mean ΔFEV_1,max?45.9 (21.9) vs. ?32.7 (22.3) %; mean difference 13.2 (3.8–22.3) %), the degree of allergen‐induced bronchial hyper‐responsiveness (mean ΔPC₂₀histamine, 1.8 (1.0) vs. 1.2 (0.9) doubling dose; mean difference 0.6 (0.2–1.1) doubling dose) and serum IL‐5 at 6 h were found to be significantly higher after bronchial challenge with HDM extract than after challenge with an isolated HDM major allergen. Likewise, there was an increased late skin reaction with HDM compared with isolated major allergen after a similar early skin reaction. Conclusion Constituents of HDM extract, other than Der p 1 or Der p 2, with no significant influence on the IgE‐mediated early asthmatic response contribute significantly to the allergen‐induced late asthmatic response and bronchial hyper‐reactivity.     

Effect of disodium cromoglycate on airway mucus secretion during antigen-induced late asthmatic responses in a murine model of asthma

BACKGROUND: Disodium cromoglycate (DSCG) is known to inhibit both immediate and late asthmatic responses (IAR and LAR). However, its effect on mucus hypersecretion is unknown. Using a murine model of asthma, we aimed to determine whether mucus secretion increased during IAR and LAR. We also studied the potency of DSCG in inhibiting mucus secretion and on airway eosinophilia. METHODS: Mice were subjected to initial intraperitoneal sensitization and airway challenge to ovalbumin (OVA) and then provoked by additional exposure to OVA. Some mice were pretreated with aerosolized DSCG (20 mg/ml) 1 h before the provocation with OVA. After serial measurements of enhanced pause (Penh), an indicator of airflow obstruction, serum samples and bronchoalveolar lavage fluids (BALF) were collected. Then, the lungs were excised and a morphometric analysis for mucus hypersecretion was performed. RESULTS: A biphasic increase in Penh (IAR and LAR) was observed in sensitized animals after provocation with OVA. Airway eosinophilia was observed during both responses. Intraluminal mucus significantly increased during LAR, but not during IAR. DSCG significantly attenuated both IAR and LAR, and significantly inhibited the increase in intraluminal mucus during LAR, but had no effect on eosinophilia in BALF. CONCLUSION: Our results suggest that airway hypersecretion may be involved as a component of airflow obstruction during LAR, and that this is unlikely during IAR. DSCG may be effective in reducing excessive airway mucus caused by exposure to allergens.     

A comparison of the effects of oral cetirizine and inhaled beclomethasone on early and late asthmatic responses to allergen and the associated increase in airways hyperresponsiveness

Background Cetirizine is a non-sedating H₁ antihistamine which is effective in the treatment of allergic rhinitis and urticaria. It inhibits eosinophil and basophil chemotaxis in late cutaneous allergic reactions in skin windows. Its effect on early (EAR) and late asthmatic reactions (LAR) is less certain. Methods We examined the effect on EAR and LAR of 3 days treatment with oral cetirizine (15mg twice daily) compared with a single dose of inhaled beclomethasone 10min prior to allergen challenge in a placebo-controlled (oral and inhaled) doubleblind cross-over design with three treatment arms separated by 14 days. Results Cetirizine did not significantly inhibit either the EAR or LAR documented by maximum percentage fall in FEV₁ (0-3 and 6-9h) or as area under the curve (AUC between 0 and 3 and 6–9 h), Beclomethasone inhibited the LAR compared with placebo (P = 0.02) when expressed as AUC (6–9h). This did not quite reach statistical significance (P = 0.06) when expressed as maximal percentage late fall in FEV₁ between 6 and 9h. A greater than twofold increase in airways responsiveness to methacholine was observed 3h after challenge which was significantly reduced by beclomethasone compared with placebo (P < 0.02) and cetirizine (P < 0.05). The data suggest that oral cetirizine does not significantly inhibit either the EAR or LAR. Beclomethasone inhibited both the early increase in airways responsiveness and the subsequent LAR. Our study also confirms the view that early increases in airway responsiveness precede the late response and suggests that these associated events are not dissociable by the pharmacological treatments employed in this study.     

Inter-Relation of Immediate and Late Asthmatic Reactions in Childhood

Following bronchial provocation tests with inhalent allergens, late asthmatic reactions (LAR) frequently follow immediate asthma reactions (IAR). This study of atopic asthmatic children demonstrated that patients could show IARs to one allergen, ryegrass extract, but isolated LARs without any preceding immediate response when challenged with an unrelated allergen extract of D. pteronyssinus. The patients' degree of skirt sensitivity to the concentration of extract inhaled appeared one factor which determined whether an isolated LAR or IAR followed allergen inhalation,     

Blood markers of early and late airway responses to allergen in asthmatic subjects. Relationship with functional findings

We evaluated the relationship between blood markers of mast-cell (plasma histamine and serum level of heat-stable neutrophil chemotactic activity [NCA]) and eosinophil (serum eosinophil cationic protein [ECP]) activation during early airway response (EAR) and late airway response (LAR) to allergen inhalation in 24 asthmatic subjects. After EAR, 14 subjects showed significant LAR (FEV₁ fall: 25%), while 10 subjects showed equivocal LAR (FEV₁ fall: 15–20%). A significant increase from baseline value was observed in plasma histamine and in serum NCA during both EAR and LAR, while serum ECP significantly increased only during LAR. The sensitivity of different markers to detect significant FEV₁ fall during EAR and LAR was low, except for NCA. Changes in blood mediators were similar in both groups with significant and equivocal LAR. There was a significant relationship between the increase in NCA during EAR and the severity of LAR. Stepwise regression between changes in different blood markers showed a significant relationship between histamine increase during EAR and ECP increase during LAR. Thus, serum NCA is a more sensitive marker of EAR and LAR than plasma histamine and serum ECP, and its increase during EAR seems predictive of the severity of the subsequent LAR.     

Early increase in urinary leukotriene E4 (LTE4) is dependent on allergen dose inhaled during bronchial challenge in asthmatic subjects

BACKGROUND: Urinary leukotriene E4 (LTE4) excretion is a good marker of the rate of total body production of sulfidopeptide leukotrienes released during allergen challenge. METHODS: Twenty-three subjects with allergic asthma were challenged with inhaled allergen, and the urinary excretion of LTE4 was determined by immunoenzymatic assay (associated with HPLC separation) at various intervals after challenge. RESULTS: Allergen challenge caused an early airway response (EAR) with a drop in FEV1 of 40.3+/-9.9%. This was associated with an increase in urine LTE4 excretion for 0-3 h after allergen inhalation (296+/-225.25 pg/mg creatinine) in comparison with baseline values obtained during the night before challenge (101.02+/-61.97 pg/mg creatinine). Urinary LTE4 excretion was significantly higher in subjects who inhaled a higher dose of allergen during challenge (LTE4 during EAR: 211+/-192 pg/mg creatinine in subjects with inhaled total dose of allergen <0.1 biologic units; 408+/-223 pg/mg creatinine in subjects with inhaled total dose >0.1 biologic units). All subjects showed a late airway response (LAR) to allergen of different severity, from mild (FEV1 fall: 15-20%) to severe (>30%); no correlation was found between the increase in urine LTE4 excreted during LAR (3-7 h after challenge) and the severity of LAR, but only subjects with severe LAR showed a significant increase in LTE4 during LAR in comparison with baseline value. CONCLUSIONS: A release of sulfidopeptide leukotrienes, as evaluated by urinary LTE4 excretion, can be documented during EAR and LAR to allergen in relation to the dose of inhaled allergen, and it can represent a useful index of the events underlying the airway inflammatory responses during allergen challenge.     

Dislocation of E-cadherin in the airway epithelium during an antigen-induced asthmatic response

The airway epithelium plays a critical role in asthma. E-cadherin, located on the basolateral side of the epithelial cells, forms adherent junctions. To investigate the role of E-cadherin on the regulation of permeability of molecules and fluid in asthmatic responses, we observed the dynamics of E-cadherin after an immunochallenge against guinea pigs. Immunohistochemical studies revealed that E-cadherin was expressed on the lateral sides of epithelial cells before the immunochallenge and after immediate airway responses (IAR). However, E-cadherin immunoreactivities decreased from the basolateral region in late airway responses (LAR) 6 h after the challenge. Simultaneously, soluble E-cadherin immunoreactivities were detected in lavage fluid only in LAR, suggesting that E-cadherin is partly cleaved and released into the lumen in LAR. Airway permeability, which was examined by penetration of horseradish peroxidase from the airway side into the epithelium, increased in both IAR and LAR. These results suggest that E-cadherin detachment from the lateral side of the epithelial cells increased airway permeability in LAR but not IAR. We conclude that an antigen challenge causes an opening of adherent junctions as well as increases airway permeability in LAR. This mechanism would participate in airflow limitation during attacks and the increase of airway permeability and hyperresponsiveness in asthmatics.