Nasal mucosal hypersensitivity in guinea pigs intermittently exposed to cold.

To develop an animal model for experimental nasal hypersensitivity and hyperreactivity, guinea pigs were subjected to intermittent exposure to cold temperature (intermittent cold stress, SART stress) for 5 consecutive days. In SART-stressed guinea pigs, nasal mucosal hypersensitivity to histamine evoking sneeze response and nasal hypersecretion in response to methacholine were observed. The hypersensitivity remained for further 7 days after being released from SART stress. On the other hand, such nasal mucosal hypersensitivity was not caused by a continuous cold stress alone, suggesting that intermittent exposure to cold may be of importance for the appearance of nasal mucosal hypersensitivity. In passively sensitized SART-stressed guinea pigs, the quantity of nasal secretion induced by an allergen was significantly increased compared with that of a group of normal animals. The expression of muscarinic acetylcholine receptor (m-ACh.R) became higher in SART-stressed guinea pigs. Thus, hypersensitivity and hyperreactivity in this system were found to be associated with an increase in density of m-ACh.R. SART-stressed guinea pigs will serve as an animal model for hypersensitivity in nasal mucosa, which would be useful for the study of nasal allergy.     

Nasal provocation with histamine in allergic rhinitis patients: clinical significance and reproducibility

In a selected group of rhinitis patients (n = 12) with an IgE-mediated allergy to house dust mites, the nasal response to insufflation of histamine chloride appeared to be related to symptom scores obtained from the patients. In contrast to the sum of the nasal airway resistances (NAR) induced by all doses of histamine, the total amount of secretion and total number of sneezes could be predicted from clinical scores. The reproducibility of the nasal provocation test was tested by comparison of the test results in two sessions with a 1-week interval. The correlation between both sessions was highest with respect to nasal secretion (r = 0.87; P less than 0.001) and the number of sneezes (r = 0.76; P = 0.004). The correlation coefficient was 0.71 (P = 0.01) when the nasal airway resistance was used in the assessment of nasal response. A good reproducibility of the nasal provocation test was also obtained using an end-point titration method and determining the concentration required to produce 0.5 ml secretion and/or five sneezes as the end-point (r = 0.76; P = 0.004). The concentration required to double nasal airway resistance yielded a correlation coefficient of 0.56 (P = 0.052). We conclude that the clinical significance of nasal provocation with histamine increases when, besides nasal airway resistance, the amount of secretion and the number of sneezes is used in the assessment of the nasal response.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunoglobulins in nasal secretion with special reference to IgE. I. Methodological studies.

Different ways of collecting relatively large volumes of nasal secretion with as physiological a composition as possible were studied. Nasal secretion was collected by the so-called nasal spray washing method from 5 patients with allergic rhinitis due to pollen and 5 healthy persons during a pollen-free season. The collection was performed without any provocation and following nasal provocation with histamine or allergen solution. With the radioimmunosorbent test, in which the lower limit of measurement was 0.1 units/ml, IgE could be quantified in 52 of 60 analysed secretions. IgA, IgG and albumin were demonstrated in all secretions. In the allergic patients, following histamine and allergen provocation, a relative increase in the concentration of IgE and albumin and a significant decrease of the IgA/albumin ratio in nasal secretion was found. In the healthy subjects, such changes in the secretion were observed only after histamine provocation. Calculations also suggested some local production of IgE, but not at all of the same order of magnitude as of IgA.     

Mechanism of nasal secretion mediated via nerve reflex in guinea pigs and evaluation of antiallergic drugs.

In order to confirm the mechanism of nasal secretion mediated via a nerve reflex in guinea pigs, the secretory response from the contralateral side was studied which was induced by local application of various stimulators. There was no difference in the nasal secretion between the contralateral and the stimulated sides when the secretion was induced by allergen, histamine, and capsaicin at lower doses. Methacholine caused a nasal secretion only on the stimulated side. Pretreatment with local anesthetic and ganglionic blockers blocked the secretory response bilaterally which was induced by allergen, histamine, and capsaicin. Antihistaminics also blocked the secretory response induced by allergen and histamine on both sides, but not the capsaicin-induced nasal secretion. Unilateral pretreatment with local anticholinergics prevented all secretory responses only on the stimulated side. Thus, exogenous and endogenous histamine released by the allergen-antibody reaction may stimulate histamine H1 receptors located in the sensory nerve endings as trigger, resulting in the secretory response mediated via a nerve reflex, while methacholine may act directly on nasal glands. Ketotifen and azelastine, which are chemical mediators releasing inhibitor with antihistaminergic activity, prevented the nasal secretion induced by histamine and allergen. On the other hand, disodium cromoglycate, amlexanox, and tranilast had only a slight effect on the allergen-induced nasal secretion. The secretory response on the contralateral side induced by various stimulators would be useful in the in vivo evaluation of anti-allergic drugs to demonstrate the difference in their modes of action.     

Histamine and methacholine do not increase nasal reactivity

Abstract. Allergen provocation in the nose increases the non-specific nasal reactivity. The aim of this trial was to determine whether this'priming effect' can be caused by histamine or methacholine, which is the most important biochemical mediator of allergic rhinitis, and an analogue to the important neurotransmittor, acetylcholine, respectively. Intranasal provocation tests with the two substances were carried out on thirteen normal subjects, and repeated 1 hr and 1 day later. The response, measured as the number of sneezes, the amount of blown secretion and the increase in nasal airway resistance, did not change with consecutive provocations. It was concluded that neither histamine nor methacholine were responsible for the allergen-induced'priming' of the nasal mucous membrane.     

Inhibition of different dosages of oxatomide or placebo on skin prick test and nasal allergen provocation.

In this two-stage, double-blind study, we evaluated the effects of different dosages of oxatomide (1 and 2 mg/kg/day) on nasal provocation and skin reaction wheal induced by grass-pollen challenge. Children with a positive history of allergic rhinoconjunctivitis and positive responses to skin prick test and nasal provocation test to grass pollen were studied out of season. The results obtained with 1 mg/kg/day of oxatomide demonstrated no significant difference in wheal areas and nasal secretion induced by allergen challenge between treated and untreated patients. The administration of 2 mg/kg/day demonstrated a significant suppression in wheal reaction and nasal secretion induced by specific challenge.     

Nasal Challenge with Serotonin and Histamine in Normal Persons

In order to study the nasal response to serotonin, 14 normal persons, in a double-blind study, were provoked in the nose with serotonin and histamine. Itching and the number of sneezes were noted, the amount of secretion measured, and nasal airway resistance recorded by active posterior rhinomanometry. Serotonin induced significant nasal itching, sneezing and hypersecretion, similar to the effects of histamine. The effect of serotonin on nasal airway resistance, on the other hand, was slight (+ 10%) and insignificant in contrast to that of histamine in equipotent doses (+ 48%) (P less than 0.001). In conclusion, we have shown that serotonin provocation can induce a rhinitis response in the human nose. The nasal symptoms suggest an effect on sensory nerves with reflex-induced sneezing and hypersecretion, while there appears to be little direct effect on capacitance vessels. The possible role of serotonin as a mediator of rhinitis remains speculative.     

Involvement of eosinophils in the early-phase allergic reaction in a guinea pig rhinitis model

BACKGROUND: Eosinophils are found in the nasal lavage fluid (NLF) and nasal biopsies of patients with allergic rhinitis after a nasal antigen challenge, and associated not only with a late-phase allergic reaction (LPR) but also an early phase allergic reaction (EPR). Numerous studies have been carried out to clarify the participation of eosinophils in LPR or airway hyperresponsiveness. However, there has been no published report describing in detail the role of eosinophils during EPR. To better understand the involvement of eosinophils in EPR, we studied the effects of repeated antigen challenges on nasal airway responsiveness and eosinophilic inflammation in EPR using a guinea pig rhinitis model. METHODS: Nasal airway responsiveness was measured as the nasal airway resistance (NAR) after nasal antigen provocation. Eosinophilic inflammation during EPR was assessed by nasal lavage and histopathological examination using two groups of animals: those in group 1 were subjected to a sensitization pretreatment only, and those in group 2 were subjected to a pretreatment of sensitization followed by repeated nasal challenges. RESULTS: Repeated antigen challenges induced nasal hyperresponsiveness as indicated by a decrease in the antigen provocation dose and a significant increase in NAR. Furthermore, significant increases in eosinophil counts, eosinophil peroxidase (EPO) activity and protein content in NLF during EPR were observed following antigen provocation in group 2. There were significant correlations between the levels of these parameters, and albumin was the most prevalent of the proteins in NLF. Histopathological examination showed that the degree of eosinophil infiltration into the lamina propria of the nasal mucosa of the animals in group 2 was significantly and apparently higher than in group 1. Particularly, epithelial disruption and mucosal edema were significantly elevated after antigen provocation in group 2. CONCLUSIONS: These results suggest that chronic eosinophil accumulation is induced by repeated antigen challenges in the nasal tissue, and that once antigen provocation occurs, eosinophils in the tissue are activated and responsible for the amplification of EPR such as vascular permeability and mucosal edema.     

艾蒿花粉诱导豚鼠过敏反应

目的：研究艾蒿花粉粗提物（MPE）和其主要成分之一artemisia vulgaris 1（Art V1）能否诱导豚鼠过敏反应,为艾蒿花粉过敏症研究建立动物模型。方法: 艾蒿花粉粗提物或Art V1混悬于佐剂氢氧化铝凝胶中,每间隔10 d致敏豚鼠1次,共4次,然后以艾蒿花粉粗提物或Art V1抗原滴鼻诱发鼻炎症状。气道高反应性(AHR)、炎症细胞和肺组织病理的观察于Art V1气雾攻击5 d后,以乙酰甲胆碱（Mch）气雾吸入激发AHR,测定气道阻力和动态肺顺应性,计数和分类计数支气管肺泡灌洗液（BALF）中的炎症细胞,观察肺组织病理学改变。 结果: Art V1组豚鼠在抗原激发下喷嚏次数和抓鼻次数显著增加,吸入Mch后气道阻力明显增加,动态肺顺应性明显降低,Mch气雾吸入可激发AHR;MPE组豚鼠喷嚏次数有所增加,气道反应性有所升高;两组BALF和病理切片均显示明显的嗜酸性粒细胞和中性粒细胞浸润。结论: 艾蒿花粉粗提物和其主要成分Art V1均能诱导豚鼠过敏反应,Art V1致敏效果明显优于艾蒿粗提物。该模型的建立有利于过敏性疾病机制和治疗新药的研究。     

In vivo effects of transient neutrophil influx on nasal respiratory epithelial mucosubstances. Quantitative histochemistry.

Certain inhaled toxicants are known to induce mucous hypersecretion in the respiratory epithelium. This secretory change may be a direct effect of the toxicant or an indirect effect of the concomitant inflammatory response. The present study was designed to determine by quantitative histochemistry whether the influx of neutrophils through the nasal respiratory epithelium would induce significant quantitative changes in the amount of intraepithelially stored mucosubstance. F344/N rats were intranasally instilled with endotoxin to elicit a transient influx of neutrophils into the nasal epithelium. Peak intraepithelial infiltration of neutrophils was evident 6 hours after instillation. There was a concurrent quantitative decrease in stored epithelial mucosubstance at the same time after instillation. Amounts of epithelial mucosubstance returned to that measured prior to neutrophil infiltration by 24 hours after instillation, when intraepithelial neutrophils were diminishing. Rats in which circulating neutrophils were sequestered in the lungs and prevented from migrating into endotoxin-exposed nasal epithelium had no change in the quantity of stored mucosubstance 6 hours after instillation. Therefore, it is concluded that a transepithelial migration of neutrophils elicits a transient depletion of stored mucosubstances in the nasal respiratory epithelium. Whether this is due to the release of secretagogues from the migrating leukocytes or another neutrophil-related method of stimulating mucous secretion is not known.     

Humoral and Cell-Mediated Immune Responses to Dog Dander and Hair in Asthmatic Children

Dog dander and hair (DDH) specific IgA, IgG and IgE antibodies from serum samples Of 202 asthmatic children, and in nasal secretion from 4 to 15 years were measured by enzyme-linked immunosorbent assay. The results were compared with clinical history, and with allergy test (skin prick test, provocation test and RAST) using the same DDH extract. A blood sample for the in vitro lymphocyte stimulation test was obtained from 40 children, and a nasal secretion sample for analysis of the local DDH-specific IgA, IgG and IgE antibody levels was collected form 35 of them. In children of dog-keeping families, higher serum levels of DDH-specific antibodies, especially IgE antibodies, were observed if the dog had been in the home already during the first years of the child's life. The serum levels of DDH-specific antibodies, however, did not correlate with the degree of the present exposure to dogs. The serum levels of DDH-specific or total IgE, or with the results of skin prick or provocation test. The serum levels of DDH-specific IgA were highest in children who were subjectively most sensitive to dogs. Nasal levels of DDH-specific IgE correlated positively with serum specific IgE levels. The correlation was weaker between nasal and serum titers of specific IgG, and not significant between nasal and serum IgA antibody levels, Specific IgE antibody levels were higher, while specific IgA and IgG antibody levels were lower, in nasal secretion form subjects with nasal symptoms on contact with dogs, when compared with subjects with other complaints (asthma, conjunctival or skin reactions). DDH-specific IgG levels correlated negatively with specific IgE level in the nasal secretion from subjects with a positive provocation test result, while the correlation was positive in subjects with a negative provocation test. The in vitro lymphocyte response to DDH did not correlate with the results of allergy test, or with the levels of DDH-specific antibodies in serum or in nasal secretion.     

Methodological Aspects of Nasal Allergen Challenges Based on a Three-Year Tree Pollen Immunotherapy Study

During 3 years of immunotherapy with tree pollen extracts, 31 patients were provoked annually. Changes in nasal reactivity were followed by registration of expiratory nasal peak flow, number of sneezes, and amount of secretion. The reproducibility of the peak flow measurements was studied. The results from all three parameters were used to form a total nasal provocation score which, better than each parameter separately, could demonstrate the variation in sensitivity. Provocation with an allergen concentration of 1 HEP was the most effective means of showing changes in specific sensitivity of nasal mucosa.     

Inhibition of allergen-induced bronchoconstriction in sensitized guinea pigs by orally administered allergen

Crude mite extract (CME) was orally administered to guinea pigs sensitized to CME. It was shown that such treatment reduces the bronchoconstrictive response upon allergen provocation. Isolated tracheae taken from guinea pigs orally administered CME allergen showed less contraction in response to CME as compared to those obtained from sensitized but not orally treated animals. The oral administration of allergens seemed to attenuate the bronchial hyperresponsiveness of sensitized animals to a non-specific chemical stimulus (histamine). IgE antibodies titrated by 8 days passive cutaneous anaphylaxis, and IgG1 and IgG2 antibodies measured by ELISA were comparable in the sera obtained from animals before and after CME treatment.     

Human nasal mucosal carboxypeptidase: Activity, location, and release

Background: Carboxypeptidases (CPs), such as carboxypeptidase N (CPN) (kininase I, E.C.3.4.17.3), may regulate peptide-mediated vasodilation and vascular permeability in respiratory mucosa by degrading proinflammatory peptides such as bradykinin, anaphylatoxins, and neuropeptides during allergic and nonallergic inflammation. The sources of CP activity in human nasal secretions were investigated. Methods: Well-characterized human nasal provocation and secretion analysis methods were used. Potential sources of CPN in human nasal mucosa were identified by immunohistochemistry. CP activity was defined as DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid inhibitable Bz-Gly-Lys degradation. CP activity was measured in nasal mucosal homogenates and nasal lavage fluids induced by methacholine, histamine, and allergen nasal provocation. Results: CPN-immunoreactive material was localized to the glycocalyx of the epithelium, some vessels, and gland ducts near the epithelial basement membrane but not to submucosal gland cells. CP activity in human nasal lavage fluid after saline nasal provocation was 0.10 ± 0.04 U/L. Histamine provoked secretion of significantly more CP activity (3.84 ± 0.99 U/L; p < 0.01 vs saline). Methacholine did not significantly increase secretion (0.54 ± 0.22 U/L). After nasal allergen challenge, CP activity was at a maximum between 11 and 20 minutes, and CP activity correlated with IgG concentration (r = 0.91, p < 0.01), a marker for proteins of plasma origin, suggesting that CP activity originated in plasma. Conclusions: These data suggest that plasma is the predominant source of CP activity secreted from human nasal mucosa and that plasma extravasation and interstitial fluid exudation across the epithelium are the primary processes regulating its appearance in nasal secretions. (J ALLERGY CLIN IMMUNOL 1995;96:924-31.)     

Nedocromil sodium reduces allergen-induced plasma extravasation in the guinea pig nasal mucosa by inhibition of tachykinin release

The effect of topically applied (10 μl) nedocromil sodium (NS) and sodium cromoglycate (CS) on the plasma extravasation induced by local application of ovalbumin (5%, 10 μl) into the respiratory nasal mucosa of sensitized guinea pigs pretreated with the neutral endopeptidase inhibitor, phosphoramidon, was studied. Topical NS (220 nmol, 10 μl) reduced by 57% the Evans blue dye extravasation caused by local application of ovalbumin into the nasal mucosa, whereas CS (220 nmol, 10 μl) was without effect. The tachykinin NK₁ receptor antagonist CP-99994 (2 μmol/kg, i.v.) reduced by 45% the plasma extravasation induced by antigen challenge. The combination of NS and CP-99994 did not increase further the inhibition caused by NS alone. Plasma extravasation evoked by instillation of bradykinin (50 nmol), which causes this response by releasing tachykinins from sensory nerves, was markedly reduced by NS, but not by CS. Plasma extravasation evoked by instillation of substance P, which acts directly on the endothelial cells, was not affected by NS. We conclude that the reduction by NS of the plasma extravasation induced by antigen challenge in the nasal mucosa of sensitized guinea pigs is due to the inhibition of tachykinin release from sensory nerve endings.     

Histamine content, synthesis and degradation in nasal mucosa and lung of guinea-pigs treated with toluene diisocyanate (TDI)

We have reported the presence of a histamine synthesizing enzyme, histidine decarboxylase (HDC), and histamine degrading enzymes, histamine N-methyltransferase (HMT) and histaminase (diamine oxidase, DAO) in human nasal mucosa and the histamine content of the mucosa. In this study, we demonstrate the influences of the toluene diisocyanate (TDI) treatment on the histamine content and these enzyme activities in guinea-pigs as an animal model of respiratory hypersensitivity. Application of TDI to the nasal vestibuli induced intense nasal allergy-like and mild asthma-like responses in TDI-sensitized guinea pigs. Increases in the histamine content and HDC and HMT activities were observed in the nasal mueosa and lung of TDI-sensitized guinea pigs. No apparent changes in the histaminase activities were observed in either the nasal mucosa or the lung. These data suggest that the turnover rate of histamine is increased in the nasal mucosa and the lung of guinea pigs with respiratory hypersensitivity.     

Facial thermography during nasal provocation tests with histamine and allergen

Changes of skin temperature (T°) of the nose area during nasal provocation tests with histamine and allergen were followed by means of an infrared thermography camera. By a colimator system in which temperatures measured on a given surface can be integrated and averaged, thermography allows the continuous and quantitative recording of the temperature during the whole procedure in a completely noninvasive way. In 10 normal subjects, increasing doses of histamine induced a dose-dependent rise of the nose external temperature. No significant change was observed with the vehicle solution. In six subjects allergic to grass pollen, the nebulization of increasing concentrations of a pollen extract induced a dose-dependent rise in T°. The T° rise observed after histamine or allergen corresponded to a marked nasal obstruction. The nebulization of the highest dose of the pollen extract did not induce any T° rise in six nonallergic subjects. The continuous recording of the skin temperature by a noninvasive method might yield additional information on the vascular changes rapidly occurring during nasal challenges.     

Effect of ion transport inhibitors and methacholine on short-circuit current of isolated guinea pig nasal epithelium

To clarify the ion/water secretion mechanism in the nasal epithelial cells, the Ussing chamber method was applied to the nasal mucosa isolated from guinea pigs. The preparation, which contained surface epithelial cells, showed a small but consistent potential difference between mucosal and submucosal sides (mucosal surface negative to submucosa). The short-circuit current (Isc) across the epithelial layer was measured, and the effects of Na+ and/or Cl- transport inhibitors and methacholine (MCh) on Isc were analyzed. The basal Isc was almost totally suppressed by the combined application of amiloride (Na+ transport inhibitor) and low-Cl- Krebs Ringer (KR) solution or solutions containing Cl- transport inhibitors (furosemide or DPC). The application of MCh elicited triphasic Isc responses, i.e., initial transient increase (phase 1) followed by a small decrease (phase 2) and further sustained increase (phase 3) in Isc. A possible ionic mechanism underlying phase 1 and 3 responses was analyzed. The Phase 1 response was greatly reduced by low-Cl- KR solution or furosemide but not influenced by amiloride. The Phase 3 response was augmented by amiloride and suppressed by low-Cl- KR solution, furosemide or DPC. These findings indicated that the basal Isc was associated with Cl- secretion and/or Na+ absorption across epithelial cells under short-circuit condition and that MCh increased Isc probably via enhancing Cl- secretion in the nasal surface epithelial cell.     

两种方法描记谷物粉尘所致豚鼠哮喘模型呼吸曲线的对比观察

为了研究动物哮喘模型的阳性判定标准,本文观察分析了对同一批哮喘模型用两种不同方法描记的激发前后呼吸曲线。结果发现呼吸频率和潮气量不能非常准确地反映哮喘发作。肺总阻力(R_L)和动态肺顺应性(Cdyn)则可以直接反映哮喘反应,但测定装置和方法复杂。用简易方法描记的规律性叹气频率与R_L 呈非常显著地正相关(r=0.81,P<0.01),而与Cdyn则呈非常显著地负相关(r=-0.73,P<0.01)。因此,叹气频率能间接地反映支气管哮喘发作。     

Substance P as a potent stimulator of sneeze responses in experimental allergic rhinitis of guinea pigs

Substance P was examined for sneeze-inducing activity and its involvement of sneeze responses in experimental allergic rhinitis. Substance P, dripped into a nostril of guinea pigs, at concentrations of 100 pM and above induced sneezing in a dose-dependent fashion. The activity of substance P was not affected by the previous subcutaneous injections of capsaicin that depleted substance P in nerve fibers. Histamine induced sneezing at concentrations of 30 mM and above and the activity was reduced by capsaicin treatment. The frequency of antigen-induced sneezing was proportional to the substance P content in nasal mucosa of sensitized guinea pigs treated with increasing doses of capsaicin; correlation coefficient 0.91. These results suggest that substance P plays an important role as a stimulator of sneeze responses in experimental allergic rhinitis in guinea pigs.