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Previous investigations have suggested that nasal secretions, obtained by lavage or scraping, and the nasal submucosa, sampled by biopsy, are two distinct compartments. We investigated the effect of intranasal corticosteroids on antigen-induced eosinophil influx into both compartments. We performed a double-blind, placebo-controlled study in 15 patients with seasonal allergic rhinitis. Beclomethasone dipropionate, 84 microg twice a day, was delivered to one nostril while the other nostril received placebo for 1 wk. Subjects were then challenged with grass or ragweed extracts on each inferior turbinate. Nasal scrapings from both inferior turbinates were obtained before and 24 h after challenge, and bilateral inferior turbinate biopsies were obtained 24 h after challenge, with the subjects still receiving treatment. Intranasal steroids led to a significant reduction in sneezes and eosinophil influx in nasal secretions without affecting the number of eosinophils in the submucosa. Furthermore, intranasal steroids had no effect on the numbers of submucosal EG2+ (activated eosinophils) or CD25+ (IL-2-receptor-bearing) cells, nor did they decrease the endothelial expression of vascular cell adhesion molecule-1 (VCAM-1). These data show that pretreatment with intranasal steroids successfully inhibited the clinical response to allergen and reduced eosinophils in the superficial compartment of the nasal mucosa, but it had no effect on inflammation in the deeper compartment. This might be related to a different distribution of the active medication and antigen into the nasal mucosa or to a specific effect of the active medication on the epithelium resulting in inhibited migration of eosinophils across this layer.

Allergic rhinitis affects about 20% of the US population. The diagnosis is based on patterns of symptoms, physical examination, and assessment of IgE antibodies by skin or in vitro testing. The most common offending allergens are pollens of grasses, trees, and weeds; fungi; animal allergens; and dust mites. In an individual with nasal allergy, exposure leads to rapid release of mast cell-derived mediators. This immediate response is followed by a cell-dominated response, including eosinophils and lymphocytes. Cytokines from T(H)2 lymphocytes, such as interleukin 4 and interleukin 5, orchestrate allergic inflammation. Resulting tissue changes produce symptoms of the disease and augment responses on subsequent exposure to allergens and irritants. Strategies for avoiding offending agents are important in management. In intermittent disease, antihistamines and/or decongestants are first prescribed. More continuous symptoms may mandate intranasal steroids. Immunotherapy is often helpful for patients who respond poorly to pharmacotherapy and avoidance.

The study of the IgE response to seasonal antigen exposure is limited by its occurrence once a year and by the variability of patient exposure to pollens. To overcome these problems, we investigated whether nasal challenge with antigen causes an increase in serum anti-ragweed IgE levels. We challenged individuals with ragweed allergy intranasally with nanogram quantities of ragweed antigen extract and measured their serum anti-ragweed IgE levels before and at weekly intervals after challenge. In a series of studies we found that there was a reproducible rise in antigen-specific serum IgE levels beginning the first week after challenge that plateaued at about 180% of baseline levels during the fourth week and remained elevated for 8 weeks. Not all individuals showed this response. The magnitude of the allergen-specific IgE response to nasal challenge appeared to be greater than the response to seasonal exposure. Treatment with intranasal beclomethasone before challenge did not affect the response. The results demonstrate a human in vivo model for the study of the antigen-specific secondary IgE response to allergen.

Background: Immunotherapy effectively treats the symptoms of allergic rhinitis and improves its pathophysiology. We studied whether the effects of immunotherapy on the early response to nasal challenge with antigen and seasonal symptoms persist after discontinuation., Methods: Twenty subjects with ragweed allergy who were receiving immunotherapy and who had nasal challenges performed before initiation of treatment were selected. The patients had been receiving maintenance therapy with aqueous ragweed extract at a dose of 12 microg of Amb a 1 equivalent for a minimum of 3 years, at which point they were randomized to receive either placebo injections or to continue with the maintenance dose. Nasal challenges were performed before and 1 year after randomization. Nasal challenges were monitored by counting the number of sneezes and measuring histamine, N-alpha-tosyl-L-arginine methyl ester-esterase activity, and kinins in recovered nasal lavages. In the same year symptom diaries were collected during the ragweed season., Results: The initial immunotherapy significantly reduced responses to nasal challenge in both groups. The group continuing to receive active treatment showed no significant changes from the response before randomization. In contrast, the group randomized to placebo treatment showed a partial return of histamine, kinins, and N-alpha-tosyl-L-arginine methyl ester-esterase in nasal secretions and the numbers of sneezes. IgG antibodies to ragweed declined only in the group switched to placebo treatment. Seasonal rises of IgE antibodies to ragweed did not return during the first season after treatment was stopped. Symptoms reported during the ragweed season were not different between the groups., Conclusions: One year after discontinuation of ragweed immunotherapy, nasal challenges showed partial recrudescence of mediator responses even though reports during the season appeared to indicate continued suppression of symptoms.

Studying the inflammatory response that follows the early response to nasal challenge with antigen provides a better understanding of allergic rhinitis than just studying the immediate (early) response. Nine allergic volunteers were challenged unilaterally with antigen-containing discs, and bilateral changes in physiologic responses as well as in the concentration of histamine in nasal secretions were measured for 11 h. We found significant immediate increases in symptoms, sneezes, ipsilateral nasal airway resistance, and ipsilateral histamine in the early phase response. Two-thirds of the allergen-challenged volunteers showed increases in physiologic parameters or histamine in the hours after allergen challenge. The pooled data of all subjects exhibited significant increases in bilateral nasal airway resistance and in ipsilateral and contralateral histamine, hours after unilateral provocation. These responses differed significantly from control subjects. In another group of 11 volunteers challenged ipsilaterally with antigen, the number of basophils increased both on the side of challenge and on the contralateral side. The magnitude of the increase on the ipsilateral side correlated with the increase on the contralateral side (r(s) = 0.72). The basophils are the most likely source of the contralateral increase in histamine as they are on the ipsilateral side. Although the mechanisms underlying this contralateral increase in basophils and histamine are not known, we speculate that delayed, neurogenic responses play a contributory role.

Ten subjects with asymptomatic seasonal allergy, outside of their allergy season, underwent allergen provocation following 1 hour of exposure to air at either 20 degrees C and 30% relative humidity (RH) or air at 37 degrees C and 90% RH. The ipsilateral changes following antigen challenge were compared under the two conditions. Conditioning of the nose to 37 degrees C, 90% RH reduced total histamine release (7.9 +/- 1.8 ng vs 4.2 +/- 1.3 ng; p < or = 0.05), sneezes (6 +/- 2 vs 3 +/- 1; p < or = 0.05), pruritus (score of 17.4 +/- 6.0 vs score of 2.0 +/- 1.8 out of a total score of 100, p < or = 0.01), nasal airway resistance (1.4 +/- 0.8 kPa/L/sec vs 0.2 +/- 0.1 kPa/L/sec; p < or = 0.05), human serum albumin levels (389.6 +/- 53.4 micrograms vs 242.2 +/- 37.9 micrograms; p < or = 0.05), and congestion (score of 23.8 +/- 4.8 vs score of 10.6 +/- 5.4 out of a total score of 100, p < or = 0.01). It had no effect on the volume of secretions (p = 0.8), lactoferrin levels (p = 0.3), or rhinorrhea (p = 1.0). Thus air at 37 degrees C and 90% RH partially reduces the early response to antigen. Its effects are greatest on histamine release, the vascular response, and neural responses, with no effect on the glandular response. The mechanisms underlying these effects are unknown.

To understand the events involved in selective eosinophil migration into allergic inflammatory sites, we studied the expression of vascular endothelial adhesion molecules in the nasal mucosa. Ten subjects with asymptomatic seasonal allergic rhinitis and 13 nonallergic subjects underwent localized allergen challenge of one inferior turbinate. Twenty-four hours later, biopsy specimens were obtained from the inferior turbinates, bilaterally in the seasonally allergic subjects and unilaterally in the nonallergic control subjects. The specimens were divided, sectioned, and either stained for identification of eosinophils or analyzed immunohistochemically for intercellular adhesion molecule-1, E-selectin, vascular cell adhesion molecule-1 (VCAM-1), and von Willebrand's factor. Intercellular adhesion molecule-1 expression was observed in all mucosal specimens, with no significant difference among groups. E-selectin showed minimal baseline expression, and low levels were significantly induced on the challenged mucosa of the allergic compared with nonallergic subjects (p < 0.05). VCAM-1 was expressed basally and was significantly upregulated by allergen challenge, compared with the nonchallenged side and nonallergic control subjects (p < 0.05). Submucosal eosinophils increased significantly in allergic subjects 24 hours after antigen challenge, compared with nonallergic control subjects and weakly correlated with VCAM-1 expression (rs = 0.33, p = 0.06). Our results suggest that endothelial activation accompanies allergic inflammation. Furthermore, because the counterligand for VCAM-1, very late activation antigen-4, is present on eosinophils, VCAM-1 upregulation may contribute to the selective recruitment of these cells to the nasal mucosa.

Allergic rhinitis is the sixth most prevalent chronic health condition in the United States. To study the pathogenesis of the allergic response, we have used a model of nasal provocation with antigen. During the initial reaction of an allergic subject to allergen provocation, increases occur in the levels of histamine, tryptase, and prostaglandin D2. This pattern of mediator release, combined with histologic evidence of mast-cell degranulation, strongly supports the role of the mast cell in the acute allergic reaction. The response to antigen, however, does not end with mast-cell degranulation. Hours after challenge we observed the spontaneous recurrence of symptoms and increased responsiveness to antigenic and nonantigenic stimuli. Our central hypothesis is that cellular infiltration and activation after antigen challenge are responsible for the observed increase in nasal reactivity. The predominant cells in nasal lavage 24 hours after challenge are eosinophils and neutrophils, whereas the predominant cell in the mucosa is the CD4+ lymphocyte. An early step in the movement of cells from the peripheral blood involves adhesion between circulating leukocytes and the endothelium. Evidence suggests that vascular endothelial adhesion molecule may be responsible in part for the selective adherence of eosinophils to the endothelium.

We enrolled nine allergic subjects in a double blind, placebo-controlled study to examine the effect of premedication with 0.6 mg of atropine on nasal antigen challenge. The challenge consisted of unilaterally stimulating the nasal septum with diluent followed by three increasing doses of antigen and recording responses bilaterally. Sneezes, symptoms, and nasal airway resistance (NAR) were recorded. Secretions were collected using preweighed filter paper discs and histamine was measured. Antigen challenge with the subjects on placebo led to significant dose-dependent increases in sneezes, symptom scores, ipsilateral and contralateral secretion weights, ipsilateral NAR, and total amount of ipsilateral histamine (p < 0.05 versus diluent). Bilaterally applied atropine led to significant inhibition of ipsilateral and contralateral nasal secretions as well as rhinorrhea scores (p < 0.05 versus placebo) but had no significant effect on other parameters. Challenge after atropine premedication led to higher increases in histamine concentration than placebo (p < 0.01). These results suggest that parasympathetically stimulated fluids did not contain histamine and diluted the histamine released by mast cells. To support this hypothesis, we challenged the same subjects with methacholine. The concentration of histamine decreased and was significantly lower than after challenge with antigen (p < 0.01). The data suggest that: (1) histamine is released locally at the site of antigen challenge, (2) the volume of glandular secretions is primarily controlled by parasympathetic stimulation, and (3) the total amount of a mediator recovered in a fixed time interval best reflects the underlying pathophysiologic events.

To investigate the role of H1 receptor-mediated effects in allergic rhinitis, we challenged 12 allergic volunteers with allergen 2 hours after administration of either placebo or 60 mg of terfenadine. Filter paper discs were used for the unilateral administration of allergen and the collection of nasal secretions. Secretion weights, levels of histamine in recovered nasal secretions, and nasal airway resistance (NAR) were measured for each nostril separately, and the number of sneezes was counted. After placebo treatment, allergen challenge led to significant increases in ipsilateral and contralateral secretion weights, ipsilateral histamine levels, ipsilateral NAR, and sneezing. Contralateral histamine levels were not elevated. H1 antagonism with terfenadine markedly reduced the number of sneezes and partially decreased ipsilateral and contralateral secretion weights, without affecting the increase in NAR. Terfenadine premedication also lowered the amount of histamine in ipsilateral secretions after allergen challenge. Performing identical nasal challenges with a 10-fold lower dose of antigen produced similar results. Previous studies showed that terfenadine had no effect on methacholine provocation and completely abolished ipsilateral and contralateral secretion weights after histamine challenge. We conclude that sneezing after allergen challenge is caused almost exclusively by a reflex initiated through H1 receptors and that H1 antagonism has no influence on allergen-induced increases in NAR. Unilateral allergen challenge leads to bilateral increases in secretion weights, which are only partially inhibited by terfenadine, suggesting the involvement of mediators other than histamine in the nasonasal reflex. As reported earlier, terfenadine also decreases allergen-induced histamine release after challenge with the highest dose of antigen.

To evaluate changes in the nasal response to histamine, we challenged 19 subjects with allergic rhinitis caused by ragweed (RW) before, during, and after the RW season with increasing doses of histamine diphosphate. We compared their response, as measured by symptoms and the levels of TAME-esterase activity and albumin recovered in the nasal lavage fluid, with response of two groups with allergic rhinitis undergoing immunotherapy with moderate-dose (N = 16) and high-dose (N = 11) RW (2 and 24 micrograms of antigen E [Amb a I] as maintenance dose, respectively). Four challenges with histamine were performed in each group: before, at the peak of, near the end of, and 2 weeks after the RW season. The three groups of subjects had similar skin sensitivity to antigen and levels of TAME-esterase activity and albumin recovered from nasal lavages after histamine challenge performed before seasonal exposure. Symptom diaries obtained throughout the season revealed a significant reduction only in the high-dose immunotherapy-treated group. At the peak of the season, the untreated group had more symptoms in response to the challenge compared with the challenges before and after the season (p = 0.04 for both groups). The saline challenge occurring before challenging with histamine also demonstrated a significant increase at the peak of the season compared with increases before the season (p = 0.02). This observation was also true for the levels of albumin and TAME-esterase activity. If the response after saline challenge was subtracted from each response after histamine challenge, no difference was found in the results between any of the visits.(ABSTRACT TRUNCATED AT 250 WORDS)

In order to compare results of allergen immunotherapy in paediatric and adult populations, 22 children with a history of ragweed hay fever were matched with an equal number of adults for skin sensitivity to ragweed and all were given a 1-year course of immunotherapy with a partially purified ragweed extract. Biological responses were measured by nasal challenges with ragweed before therapy was started, after 12 weekly injections and when the maintenance dose had been reached and also by methacholine bronchoprovocation tests before and after 12 months of therapy. Skin-test sensitivity to ragweed and control allergens, and ragweed-specific IgE and IgG antibody responses were measured at the same intervals as the challenges and at the end of the study. The effect of the therapy on clinical symptoms was not evaluated. Before therapy the groups of adults and children were comparable by all indices, except for TAME esterase activity in nasal washes during ragweed nasal challenge which was significantly lower in children. During treatment, mediators released during sequential nasal challenges declined to undetectable levels in most patients and changes in nasal ragweed sensitivity were comparable in both groups. Ragweed IgE increases after 12 weeks of therapy and IgG levels at maintenance therapy tended to be higher in the children, but neither difference was statistically significant. At the end of the study IgE and IgG antibody levels were comparable in both groups. Results of methacholine inhalation tests did not change significantly in either group. The decrease in skin sensitivity to ragweed was similar in both groups. We conclude that ragweed immunotherapy leads to immunological and biological consequences that are comparable in children and adults.

We have developed sensitive amplified immunoassays for measurement of IgE and IgG4 ragweed (RW) antibodies in unconcentrated nasal washes. IgE to Amb a I (formerly antigen E) can be assayed to less than 0.1 ng/ml using IgE capture by anti-IgE on microtitre plates and an alkaline phosphatase-conjugated Amb a I with an amplification substrate technique. IgG4 to whole RW extract was assayed to less than 0.01 ng/ml by amplification ELISA using monoclonal anti-IgG4. Nasal washes (NW) (10 ml) and serum were obtained in December from 22 RW-sensitive patients before and after 1 and 2 yr of RW immunotherapy (IT), and assayed for Amb a I IgE or RW IgE and RW IgG4 antibodies Amb a I IgE could be measured in the NW of 15/22 pre IT, 19/22 at 1 yr IT, but only 3/10 at 2 yr IT (compared with pre-IT, P less than 0.05). Mean Amb a I IgE in NW was 0.66, 0.36 and 0.21 ng/ml at pre, 1 yr and 2 yr IT (P-values greater than 0.05). Mean serum RW IgE, was 76, 55 and 27 ng/ml at pre, 1 yr and 2 yr IT (P-values greater than 0.05). Amb a I IgE in nasal washes was correlated with RW IgE in serum (r = 0.56, P less than 0.001, n = 44). RW IgG4 was detectable in NW of 15/22 pre-IT, 18/22 at 1 yr IT and 9/10 at 2 yr IT (P-values greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Using a recently developed model of nasal challenge, we have obtained data that clearly demonstrate, for the first time, kinin generation during a local allergic reaction in vivo. Allergic individuals (n = 8) and matched nonallergic controls (n = 8) were challenged intranasally with the appropriate antigen and nasal washes were taken before and after challenge. Washes were assayed for kinin, histamine, and [3H]-N-alpha-tosyl-L-arginine methyl ester (TAME)-esterase activity. Increased kinin generation was found by radioimmunoassay (RIA) in the nasal washes of all the allergics (5,560 +/- 1,670 pg/ml) but in none of the controls (38 +/- 16 pg/ml). The presence of kinin was highly correlated with that of histamine and TAME-esterase activity and with the onset of clinical symptoms (P less than 0.001). Serial dilutions of nasal washes produced RIA displacement curves that paralleled the standard curve, and recovery of standard kinins that were added to nasal washes was 100 +/- 4% (n = 14). Kinin recovery was identical in both allergics and controls and did not vary significantly with antigen challenge. The immunoreactive kinin in nasal washes was stable to boiling and not precipitated by ethanol, but completely destroyed by carboxypeptidase B. It was evenly distributed between the sol and gel phases of nasal washes. High performance liquid chromatography analysis of the immunoreactive kinin in nasal washes showed it to be a mixture of lysylbradykinin and bradykinin. We conclude that kinins are produced during local allergic reactions in the nose and may contribute to the symptomatology of the allergic response.

During the course of an open immunotherapy (IT) study of ragweed (RW)-allergic patients, nasal mediator release was studied by provocation testing. All subjects had a history of seasonal RW rhinitis, positive skin puncture test to RW, and RW-specific IgE by RAST. Nasal challenge was performed with serial dilutions of RW extract, before and after 12 weekly injections, providing a cumulative dose of 0.22 microgram of Amb a I. Serum IgE and IgG and basophil histamine release with RW were also measured. By 12 weeks of IT, when only 1% of the usual maintenance level dose had been administered, mean histamine release and TAME-esterase activity in nasal washes decreased significantly (p less than 0.05 and p less than 0.01). Prostaglandin D2 release did not change. Skin sensitivity decreased (p less than 0.05), whereas RW-specific IgE increased (p less than 0.05). No significant change in basophil histamine release was observed for RW or a control antigen. Only six of 40 subjects had an RW-specific IgG rise greater than 0.05 microgram/ml. Changes in nasal sensitivity did not correlate with the increases in IgE or IgG or with the change in skin test sensitivity. These present data indicate that there is a significant decline in nasal sensitivity to inhaled RW very early in the course of IT. There is, however, no indication of a relationship between the decreased nasal sensitivity and the production of RW-specific IgG antibodies.

An in vivo model of human allergic disease has been developed in which nasal challenge with antigen leads to physiologic changes, together with a release of increased amounts of inflammatory mediators into nasal secretions obtained by washing the nose with saline. In 105 experiments involving 35 subjects, only allergic subjects consistently demonstrated an increase in the concentrations of the mast cell mediator, histamine, and the putative mast cell mediators, TAME-esterase and PGD2. The release of each mediator was significantly (p less than 0.001) related to the physiologic change (sneezing). The release of each mediator also correlated significantly with the release of the other 2 mediators (p less than 0.001). This system, for the first time, clearly relates an in vivo symptom and mediator release and thus should provide an excellent tool for the further study of the allergic response and nasal pathophysiology.

Background Multiple mediators including prostaglandin D2 and leukotriene B4 have been shown to increase in nasal secretions during the early response to nasal challenge with antigen. Objective Our objective was to investigate the time course of prostanoid and leukotriene B4 release into nasal secretions on both the ipsilateral and contralateral side after a unilateral nasal allergen challenge. Methods We performed a controlled, randomized trial. Six volunteers were challenged unilaterally with antigen or diluent in a randomized order and discs were used to collect nasal secretions from both nostrils at 2min intervals for 20min after the challenge. Prostanoids and leukotriene B4 (LTB4) in recovered nasal secretions were measured by combined capillary gas chromatography-negative ion chemical ionization mass spectrometry (GC/MS). Results Nasal allergen challenge resulted in a significant and immediate increase in symptoms and sneezing, PGD2 was significantly elevated above diluent values (0.6±0.6pg) 30s after removal of the allergen disc (P<0.05), reached its peak (423.2±182.4 pg) at 2min and then slowly decreased. PGD2 also increased on the contralateral side after unilateral allergen challenge, reaching peak values about six times lower than on the ipsilateral side (70.8±21.7pg at 6min). Levels of 9a, llb- PGF2 after antigen provocation became significantly higher than after diluent (0±0pg) on the ipsilateral side at 2min (17.2±5.9pg), and reached peak levels at 4 min (25.1±8.0pg). LTB4 also increased significantly on the side of challenge. For the other prostanoids measured (PGF2, PGF2α, TxB2, 6kPGF10), no significant changes in either ipsilateral or contralateral secretions were observed after allergen challenge. Conclusions Our study described the kinetics of PGD2 and LTB4 release as well as the contralateral release of PGD2. [ABSTRACT FROM AUTHOR]

Background Multiple mediators including prostaglandin D2 and leukotriene B4 have been shown to increase in nasal secretions during the early response to nasal challenge with antigen. Objective Our objective was to investigate the time course of prostanoid and leukotriene B4 release into nasal secretions on both the ipsilateral and contralateral side after a unilateral nasal allergen challenge. Methods We performed a controlled, randomized trial. Six volunteers were challenged unilaterally with antigen or diluent in a randomized order and discs were used to collect nasal secretions from both nostrils at 2min intervals for 20min after the challenge. Prostanoids and leukotriene B4 (LTB4) in recovered nasal secretions were measured by combined capillary gas chromatography-negative ion chemical ionization mass spectrometry (GC/MS). Results Nasal allergen challenge resulted in a significant and immediate increase in symptoms and sneezing, PGD2 was significantly elevated above diluent values (0.6±0.6pg) 30s after removal of the allergen disc (P<0.05), reached its peak (423.2±182.4 pg) at 2min and then slowly decreased. PGD2 also increased on the contralateral side after unilateral allergen challenge, reaching peak values about six times lower than on the ipsilateral side (70.8±21.7pg at 6min). Levels of 9a, llb- PGF2 after antigen provocation became significantly higher than after diluent (0±0pg) on the ipsilateral side at 2min (17.2±5.9pg), and reached peak levels at 4 min (25.1±8.0pg). LTB4 also increased significantly on the side of challenge. For the other prostanoids measured (PGF2, PGF2α, TxB2, 6kPGF10), no significant changes in either ipsilateral or contralateral secretions were observed after allergen challenge. Conclusions Our study described the kinetics of PGD2 and LTB4 release as well as the contralateral release of PGD2. [ABSTRACT FROM AUTHOR]