Nasal Antigen Challenge Research Articles

In vivo release of 15-hete and other arachidonic acid metabolites in nasal secretions during early allergic reactions

The purpose of this study is to examine to “in vivo” release of 15-HETE and other arachidonic acid metabolites in nasal secretions following a challenge with “Dermatophagoides Pteronyssinus” in patients with allergic rhinitis and non-allergic controls. In addition, we examine the effects of a membrane stabilizer, such a sodium cromoglycate, on these metabolites Thirteen allergic subjects and seven healthy controls are studied. 15-HETE , peptide leukotrienes, LTB 4 PGD 2, PGE 2 and PGF 2α levels are evaluated before and after nasal challenge in sodium cromoglycate treated and untreated subjects. This study provides “in vivo” evidence that the pathophysiological responses to nasal antigen challenge could be related to the release of 15-HETE as well as other arachidonic acid metabolites, mainly arising from the lipoxygenase pathway.

アレルギー性鼻炎患者におけるケミカル・メディエーターに関する研究 第二報 二相性反応の鼻汁中ヒスタミン，ロイコトリエン，キニンに関する検討

In order to clarify the details of a late phase response (LPR) in allergic rhinitis, serial changes of nasal symptoms and concentration of chemical mediators (histamine, leukotriene (LT) C4 and kinins) in nasal lavage fluid were investigated after the nasal antigen challenge in ten patients with perennial allergic rhinitis with a positive intradermal skin reaction to mite extract. Samples were collected periodically for 8 hours following the challenge. Nasal airway resistance (NAR) and clinical symptoms were also recorded serially. Three out of ten patients showed only early phase response (EPR) and the other seven patients showed both early and late phase response after the challenge. The chief nasal symptom in LPR was nasal obstruction. In EPR, concentrations of chemical mediators in the nasal lavage fluid always increased. In dual (early and late) phase responders, increases of chemical mediators in nasal lavage fluids were notable during both early and late phase responses. In EPR, there was a significant correlation between severity of sneezing attack and concentration of histamine. In LPR, increase of LTC4 level was significantly correlated with the increase of NAR. RAST score and daily nasal symptoms tended to be higher and more sever in dual phase responders compared with early phase responders. We suggest that the amounts of released chemical mediators during allergic reaction denoted the LPR in allergic rhinitis patients.

25 A comparison between antigen-induced leukocyte populations in the nasal mucosa and nasal secretions

25 A COMPARISON BETWEEN ANTIGEN-INDUCED LEUKOCYTE POPULATIONS IN THE NASAL MUCOSA AND NASAL SECRETIONS. M. Brown M.D, M.D.. M. Furin. M.D.. M. Taylor. MS, D.. Ba Marym We evaluated antigen-induced leukocyte influx into both the nasal mucosa and secretions. Eight seasonal allergic (A) and 11 nonallergic (NA) subjects underwent nasal lavage before (PRE), and lavage followed by biopsy 24 hours after, nasal antigen (ANT) challenge administered locally by a paper disc placed on one inferior turbinate. Biopsy of the site was performed immediately after the lavage in both groups. In addition, the allergic group underwent contralateral turbinate biopsy in the absence of antigen provocation at another time. Eosinophils (EOS), mononuclear cells (MONO) and neutrophils (PMN) were quantified in lavages and tissues. m MUCOSA (/mm2) A-PRE 18+13 hFlmQ E 2010+451 A-ANT *246+74 *3613+364 *89562 NA-ANT **48+37 2107+225 117:53 pgz (x103) EOS %P PMN 43+28 4732250 A-ANT 2222175 *46;18 553+180 NA-ANT *19.4 6435 **46+29 (Values represent means LSEM, *p<.O5, **p<.Ol, each, compared to the value directly above.) Allergies had more EOS than the NA in lavage, a finding not paralleled in the mucosa. After challenge, the tissue number of EOS, MONO and PMN increased significantly compared to unchallenged, but only the eos increased significantly compared to the NA. In fact, the PMN in NA-ANT were increased relative to the A-PRE, possibly reflecting nonspecific irritation caused by the placement of the antigen. Changes in cell counts did not correlate between the mucosa and the lavage. Part of the differences may relate to the small area challenged. We believe the mucosa and overlying mucus layer represent two interrelated, yet distinct, compartments.

Localized Antigen Challenge of the Nasal Mucosa

We describe a localized nasal antigen challenge and the measurement of mediators found at the same site. Eight ragweed-allergic subjects were challenged on 2 days, 1 week apart. Challenges consisted of six sequential provocations, beginning with two control challenges (diluent for antigen-phenol-buffered saline) followed by four increasing antigen doses (0.6, 6, 60, and 160 protein nitrogen units) (antigen day) or an additional four control challenges (control day). The number of sneezes and the symptom scores increased significantly with increasing antigen doses. The levels of histamine and N-alpha-tosyl-L-arginine methyl ester-esterase activity increased in the eluted secretions on the antigen day, but not on the control day. The amount of secretions collected also increased per unit of time on the antigen day. We found no significant increase in the concentration level of either histamine or N-alpha-tosyl-L-arginine methyl ester-esterase in nasal secretions on either day. We conclude that the total amount of histamine and N-alpha-tosyl-L-arginine methyl ester-esterase activity increased per unit of time while their concentration did not.

Leukotriene B 4 as a mediator of early and late reactions to antigen in humans: The effect of systemic glucocorticoid treatment in vivo

The role played by leukotriene B 4 (LTB 4) in human allergic reactions has been the subject of recent interest and speculation. To characterize further the role of this mediator, we quantitated LTB 4 levels in nasal washing by radioimmunoassay in 13 atopic subjects during immediate and late reactions after nasal antigen challenge while the subjects were taking placebo or prednisone (20 mg every 8 hours for 48 hours) in a double-blind, placebo-controlled, crossover protocol and compared these levels with levels of seven nonatopic subjects undergoing similar nasal antigen challenge. Nasal antigen challenge of atopic subjects resulted in an increase in LTB 4 levels during the immediate reaction in 10 of 13 subjects ( 9 13 with a >50% increase over baseline) with no similar increase observed in nonatopic subjects ( p < 0.05). An increase in LTB 4 levels was observed in 12 13 atopic subjects ( 6 13 with >50% increase over a second baseline) during late time periods ( p < 0.05), which was associated with an influx of neutrophils (from 65,000 ± 43,000 to 1,246,000 ± 829,000; p < 0.05). However, nonatopic subjects appeared to demonstrate a similar late increase in LTB 4 levels. High-performance liquid chromatography analysis of immunoreactive LTB 4 demonstrated that 84% of immunoreactive LTB 4 coeluted with the biologically isomer during the immediate reaction, whereas 44% to 61% coeluted with the biologically active isomer during late reactions. Steroid pretreatment had no effect on either the early or late increase in LTB 4 levels or on the neutrophil influx observed during the late reaction. In a second series of studies involving eight subjects, we demonstrated that a single 50 mg oral dose of prednisone had no effect on the release of LTB 4 from neutrophils stimulated ex vivo with calcium ionophore A23187 or serum-activated zymosan, and, furthermore, that neutrophil concentration (“density”) had a marked effect on the ability of neutrophils to metabolize LTB 4 that they synthesize. These results indicate that human allergic nasal reactions are associated with the release of LTB 4 in vivo and that these levels and neutrophil influx are not modified by systemic steroid pretreatment. Because multiple inflammatory cells may participate in these allergic reactions to synthesize and metabolize LTB 4, this mediator may play a complex role in human allergic reactions.

抗原誘発後の鼻粘膜ＳＲＳ‐Ａ含有量の経時的変化

We evaluated the time course of production of SRS-A in the nasal mucosa after antigen challenge in the guinea pigs with nasal allergy and the effect of Ketotifen sprayed topically over the nasal mucosa on SRS-A production. Fifty-six Hartley strain guinea pigs were sensitized passively by intravenous injection of 0.4 ml of sera containing antiovaTbumin IgE and γ1 antibody (7 days' PCA titer, ×1664; 48 hours' RCA titer, ×5632). After 7 days, all animals received nasal challenge with 50 μl×2 of 1 % ovalbumin. Thirty-two animals were subdivided into 8 groups that were killed before the challenge and 15 min, 30 min, 1 hr, 3 hr and 12 hr after antigen challenge. The nasal mucosa was removed and the amount of LTC4, LTD4 and LTE4 in the nasal mucosa was measured using high-powered liquid chromatography combined with radioimmunoassay. In 12 animals, nasal spray of 0.1 mg Ketotifen dissolved in normal saline was given once a day for 6 days until the day of antigen challenge and another 12 animals were given nasal spray of normal saline as a control. Thirty minutes after nasal antigen challenge, the animals were killed and the nasal mucosa was removed to measure the amount of SRS-A. LTC4, LTD4 and LTE4 in the nasal mucosa increased significantly between 15 and 60 min after antigen challenge and then decreased markedly. Ketotifen sprayed topically over the nasal mucosa significantly inhibited production of LTC4, LTD4 and LTE4 by as much as 50%.

鼻アレルギーとロイコトリエン （第２報） 抗原誘発後の鼻腔洗浄液におけるペプチドロイコトリエンおよび炎症性細胞の経時的変化

The purpose of this study is to clarify the role of peptide leukotrienes (LTs) on the onset of characteristic hyperreactive nasal symptoms of nasal allergy by observing the time course of the correlation among degrees of nasal symptoms, and by observing the amount of chemical mediators and the number of inflammatory cells in the nasal lavage fluid after nasal antigen challenge in subjects with Japanese cedar pollinosis during off season. Sneezing was terminated within 10 minutes and nasal discharge within 2 hours. However, time course change of the percent increase of nasal airway resistance showed dual response consisting of immediate and late phase responses. The peak of the former was seen at 30 minutes and the latter was at 7 hours after provocation. The significant increase of eosinophils in the nasal lavage fluid was observed during both the immediate and the late phase responses, but during the late phase response, the increase was more prominent. Basophilic cells definitely increased during the late phase response. The amount of LTs in the nasal lavage fluid increased significantly during both the immediate and the late phase responses. In contrast, the level of histamine increased significantly only during the immediate phase response. Considering that LTs, especially LTD4, has potent and persistent effect on causing swelling of nasal mucosa, LTs may play important role in causing nasal obstruction during both the immediate and the late phase responses after antigen challenge. On the other hand, the role of histamine may be confined to cause the hyperreactive nasal symptoms during the immediate phase response.(ABSTRACT TRUNCATED AT 250 WORDS)

The Influx of Inflammatory Cells into Nasal Washings during the Late Response to Antigen Challenge: Effect of Systemic Steroid Pretreatment

Previous studies have demonstrated symptoms and mediator release occurring as long as 11 h after nasal challenge with antigen in selected allergic subjects. Pretreatment with systemic steroids reduced symptoms and mediators including histamine, TAME-esterase activity, and kinins. The aims of the present study were to characterize the cell influx during the late-phase response to antigen challenge and to determine the effect of pretreatment with systemic steroids on this response. We examined cytospin slides of nasal washings obtained before and hourly for 11 h after nasal antigen challenge in 10 asymptomatic allergic subjects with a history of seasonal rhinitis and 5 normal, nonallergic subjects. Allergic subjects received oral prednisone (20 mg 3 times a day) or placebo in a random, double-blind crossover manner for 2 days before each of 2 challenges 1 month apart. On placebo days, a mixed cell influx occurred in allergic subjects during the late response that was 50-fold greater than the cell influx in the nonallergic control subjects (p less than 0.005). During the first 3 h after antigen challenge, eosinophils (p less than 0.005), but not neutrophils or mononuclear cells, were observed. During the late phase (4 to 11 h), neutrophils, eosinophils, and mononuclear cells were all increased. Oral steroid pretreatment blocked the influx of eosinophils (p less than 0.005), but not that of other cells. These data demonstrate an inflammatory cell influx associated with the nasal late-phase response and suggest an important pathogenetic role for the eosinophil.(ABSTRACT TRUNCATED AT 250 WORDS)

Basophil influx occurs after nasal antigen challenge: Effects of topical corticosteroid pretreatment

Both the pattern of mediator release during the late-phase response (LPR) and the reduction of the LPR with corticosteroid pretreatment have suggested that basophils, not mast cells, represent the main source of histamine in the late response to nasal antigen challenge. We tested this hypothesis by examining alcian blue-stained cytospin slides of nasal washings obtained before and for 11 hours after nasal antigen challenge in 11 asymptomatic subjects with seasonal allergic rhinitis. In a double-blind manner, subjects received placebo or topical flunisolide (50 μg, each nostril, twice daily) for 1 week before antigen challenge. One month later, the challenge was repeated with the alternate pretreatment. On placebo-treatment days, a twelvefold increase occurred in the number and a threefold increase in the percentage of alcian blue-stained positive cells in nasal washings in the LPR compared to baseline. At least 68% of these alcian blue-stained positive cells were basophils, as determined by light microscopic criteria. Alcian blue-stained cell influx correlated with increases in histamine levels in nasal washes (p < 0.001). Topical steroid pretreatment blocked the influx of alcian blue-stained positive cells, as well as other inflammatory cells, including eosinophils, neutrophils, and mononuclear cells. Symptoms and mediator release were also blocked. These data demonstrate an influx of basophils and suggest that these cells are responsible for the histamine release observed in the LPR. Our findings indicate that pharmacologic control of basophil histamine release may represent a strategy for the treatment of a variety of chronic allergic diseases that are believed to resemble the LPR.

Eustachian tube obstruction after provocative nasal antigen challenge.

To test if allergic rhinitis is associated with abnormal eustachian tube function (ET), juvenile rhesus monkeys were passively sensitized with serum samples from patients allergic to timothy grass or ragweed pollen. Tympanostomy tubes were inserted for direct measurement of ET function by inflation-deflation and forced-response testing before and after provocative intranasal challenge. Intranasal challenge with pollen (antigen) resulted in rhinorrhea and nasal obstruction. Slight increases in passive ET function values and significantly impaired active ET function were observed after antigen challenge. No changes in nasal airway patency or active ET function resulted from control nonantigen pollen challenges. These data complement our studies in man and support the use of this model to evaluate the relationships among allergic rhinitis, ET function and middle ear disease.

Nasal conductance and effective airway diameter.

1. Transnasal pressure (delta p) in Pa and gas flow in cm3 s-1 were measured in men breathing gases of different density, assuming flow through each nostril (f) to be half the measured total flow. Measurements were also made in children with allergic rhinitis before and after nasal antigen challenge. 2. Flow always showed evidence of turbulence when transnasal pressure exceeded 40-80 Pa breathing air and Reynolds number exceeded 2400 (1800 in the presence of nasal obstruction). 3. Changes in effective mean nasal airway diameter (D) after nasal challenge can be determined at points of similar pressure from the relation log (D2/D1) = 0.368 log (f2/f1) when flow is turbulent. 4. Absolute estimates of airway diameter in cm can be obtained from the relation 4.75 log D = 1.75 log f--log delta p+log L--4.756 breathing air when flow is turbulent (since the relation between pressure and flow approximates to that found in a long cylinder) and these estimates are only marginally affected by differing assumptions about nasal airway length (L). 5. Because pressure and flow are non-linearly related, and changing nasal dimensions have a profound effect on the flow at which turbulence occurs, it is suggested that measures of nasal conductance at delta p greater than or equal to 0.1 kPa are preferable to the more conventional measures of nasal resistance at a specified flow rate.

679 SEROUS OTITIS MEDA: EVIDENCE FOR AN ALLERGIC MECHANISM

The importance of allergy in the pathogenesis of serous otitis media continues to be controversial. Our aim is to clarify the role of atopy in the development of serous otitis and to describe a technique for the further study of this relationship.Nasal challenges were performed with both diluent and rye grass pollen extract in 43 children with allergic rhinitis. Nasal airway resistance was measured by anterior rhinomanometry and nearly simultaneous tympanometry was utilized to detect changes in middle ear pressure. Fifteen of the patients had significant changes in middle ear pressure of minus 35 mm H2O or greater in one or both ears (mean=minus 60 mm). The control diluent challenges in the same group resulted in a mean change of plus 20 mm H2O (p<0.001).Significant obstructive increase (≥50%) in nasal airway resistance was observed in 32 of 43 patients (74%). Negative shift in middle ear pressure was seen in 28 of these 32 patients (88%) as compared to only 2 of 11 patients (18%) with a nasal airway resistance increase of less than 50%. All 15 patients with significant middle ear pressure changes had a nasal airway resistance increase of ≥50%.These results indicate that allergic rhinitis patients who react to nasal antigen challenge demonstrated concomitant significant changes in middle ear pressure. Persistent negative pressure in the middle ear can produce an effusion characteristic of serous otitis media. We interpret this data to indicate that allergic children have increased susceptibility to serous otitis media.