bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction, distribution and modulation of upper airway allergic inflammation in mice
Authors: Hussain I. , Randolph D. , Brody S. L. , Song S.‐K. , Hsu A. , Kahn A. M. , Chaplin D. D. , Hamilos D. L. 
Published: March 2003

Background To further elucidate mechanisms of human allergic rhinosinusitis, we studied the induction, distribution and modulation of allergen‐induced upper airway inflammation in a BALB/c mouse model.Methods Allergic inflammation induced with ovalbumin (OVA) by intraperitoneal (IP) injection in alum was compared to repeated intranasal instillation. The type and distribution of inflammatory cells was compared in the respiratory and olfactory epithelial compartments. Eosinophil distribution was assessed using Scarlet Red stain and a polyclonal antibody recognizing eosinophil major basic protein (MBP). The role of interleukin (IL)‐5 in upper airway inflammation was tested by administration of polyclonal anti‐IL‐5 antibody during the sensitization protocol.Results Unsensitized control mice receiving saline failed to develop upper airway eosinophil infiltration. IP OVA‐sensitized mice developed marked upper airway mucosal eosinophil infiltration after aerosol OVA challenge, whereas repeated intranasal instillation of OVA produced qualitatively similar, but less intense eosinophil infiltration. Using either sensitization protocol, eosinophil infiltration was seen in areas of the lower portion of the nasal septum, the floor and the lower lateral walls of the mid‐caudal region of the nasal cavity. Immunofluorescence staining for MBP confirmed this distribution of eosinophils but also demonstrated some eosinophils in the maxillary sinuses and in circumscribed regions of the ethmoturbinates. All areas of eosinophil infiltration were lined by respiratory epithelium. The selective infiltration of respiratory but not olfactory epithelium by eosinophils was unassociated with a measurable induction of epithelial ICAM‐1 or eotaxin expression. OVA‐induced upper airway eosinophil infiltration was found to be IL‐5 dependent, since administration of a polyclonal anti‐IL‐5 antibody (TRFK‐5) during OVA sensitization resulted in a marked modulation (80% decrease) in eosinophil infiltration in response to subsequent OVA challenge.Conclusion The mouse upper airway, specifically in areas containing respiratory epithelium, is a target for OVA‐induced allergic inflammation. This selective infiltration of respiratory, but not olfactory, epithelium is, in part, dependent upon IL‐5. This model is useful for further dissection of the inflammatory response with genetic manipulations and targeted immunological approaches.
A prospective study of the association between home gas appliance use during infancy and subsequent dust mite sensitization and lung function in childhood
Authors: Ponsonby A.‐L. , Dwyer T. , Kemp A. , Couper D. , Cochrane J. , Carmichael A. 
Published: March 2003

Background Home gas appliance use has been associated with child respiratory illness but prospective data on the relationship between infant exposure and the development of child allergic disease has not been readily available.Objectives (a) To determine if home gas appliance use is associated with increased risk of house dust mite (HDM) sensitization. (b) To examine whether any association between current home gas use and airway obstruction is influenced by HDM sensitization.Methods Design: an 8‐year follow‐up birth cohort study of children born during 1988 and 1989. Participants: a population‐based sample (n = 498) of children who participated in the Tasmanian Infant Health Survey (TIHS) and resided in Northern Tasmania in 1997 (84% of eligible children). Main outcome measures: (a) Skin prick test reaction to nine allergens, including Dermatophagoides pteronyssinus (Der p 1) and Dermatophagoides farinae (Der f 1). (b) Spirometric lung function indices, including forced expiratory volume in one second (FEV1) and forced vital capacity (FVC).Results The relative risk for home gas appliance use at 1 month of age and HDM sensitization was 1.98 (1.04, 3.79) in a cohort analysis with confounder matching. Current home gas use was also associated with HDM sensitization (ARR 1.73 (1.43, 2.76)). Current home gas use was related to a stronger (P = 0.006) reduction in the FEV1 : FVC ratio among HDM‐sensitive children (adjusted difference − 6.2% (− 10.0 to − 2.4)) than non‐HDM‐sensitive children (adjusted difference − 0.3% (− 2.5 to 1.8)).Conclusion Indoor pollutants from gas combustion may increase the likelihood of initial sensitization to HDM and play a role in the development of atopic asthma. HDM‐sensitized children may be more vulnerable to indoor pollutant‐induced airway obstruction. The ability of this study to detect such effects may partly reflect unflued gas appliance use among this sample.
Quantification in mass units of group 1 grass allergens by a monoclonal antibody‐based sandwich ELISA
Authors: Arilla M. C. , Ibarrola I. , Eraso E. , Aguirre M. , Martínez A. , Asturias J. A. 
Published: March 2003

Background Grass pollen extracts currently used for allergy diagnosis and immunotherapy are a complex mixture of proteins of which only a few have allergenic activity. Lol p 1 is one of the most important allergens in grass pollen extracts.Objectives To develop a two‐site enzyme‐linked immunosorbent assay for the quantification of Lol p 1 and other group 1 allergens from grass species, and to assess its suitability for quantifying this group of allergens.Methods Balb/c mice immunized with recombinant Lol p 1 were used for the production of monoclonal antibodies. Screening of hybridomas was performed by direct ELISA, and selected monoclonal antibodies were immobilized on ELISA plates and incubated with samples containing group 1 allergens. Bound allergens were detected by a combination of biotinylated Lol p 1‐specific monoclonal antibody and peroxidase‐streptavidin conjugate.Results The assay is based on three Lol p 1‐specific monoclonal antibodies with different epitope specificities. The optimized ELISA measured Lol p 1 concentrations ranging from 125 to 1000 ng/mL and could quantify group 1 allergen from grass species belonging to the Pooidea subfamily. The assay does not depend on anti‐sera production or availability of human sera and thus reactives can be produced in unlimited amounts.Conclusion This sensitive and specific Lol p 1 assay will be helpful both for quantifying the group 1 allergen content of Pooideae pollen extracts intended for clinical use and for studying cross‐reactivities among pollen extracts.
Venom immunotherapy induces monocyte activation
Authors: Magnan A. , Marin V. , Mély L. , Birnbaum J. , Romanet S. , Bongrand P. , Vervloet D. 
Published: March 2003

Background Venom immunotherapy (VIT) is an efficient treatment of hymenoptera venom allergy. The mechanism of VIT is based on the induction of tolerance of allergen‐specific Th2 cells. The mechanisms of this T cell modulation are unknown, and could depend on cytokines produced by other cell types such as interleukin (IL)‐12, tumour necrosis factor (TNF)‐α and IL‐10 by monocytes.Objective To assess if VIT modifies the monocyte production of IL‐12, TNF‐α and IL‐10 during the 45 first days of treatment.Methods Fourteen patients and seven controls were included. Blood samples were taken once in controls and at day (D)1, D30 and D45 of VIT in patients. Monocytes were isolated, cultured with and without lipopolysaccharide (LPS), and the culture supernatant was harvested. IL‐10, IL‐12 and TNF‐α were assayed in supernatants by ELISA.Results Baseline cytokine levels were not statistically different between patients and controls. During treatment, an increase of spontaneous monocyte production of IL‐12 and TNF‐α was observed at D15 and D45. The production of IL‐10 increased at D15 and D45 but not significantly. After LPS‐stimulation, IL‐12, TNF‐α and IL‐10 monocyte production was not modified by VIT.Conclusion VIT induces a monocyte activation characterized by a delayed overproduction of IL‐12 and TNF‐α. These cytokines could be relevant to the inhibition of Th2 cells during VIT. Therefore, VIT‐induced tolerance could depend not only on the specific action of venom antigens on T cells, but also on a secondary non‐specific action of monocytes.
Lymphoproliferative responses in cord blood and at one year: no evidence for the effect of in utero exposure to dust mite allergens
Authors: Smillie F. I. , Elderfield A. J. , Patel F. , Cain G. , Tavenier G. , Brutsche M. , Craven M. , Custovic A. , Woodcock A. 
Published: March 2003

Background Maternal allergen exposure beyond the 22nd week of pregnancy may be important in foetal T cell priming. Allergen‐specific cord blood mononuclear cell (CBMC) immunoproliferative responses without corresponding bacterial antigen responses (tetanus toxoid), have been suggested as evidence of in utero sensitization.Objectives To investigate the relationship between lymphoproliferative responses at birth and at 1 year with maternal and 1‐year infants house dust mite allergen exposure.Methods Home visits and dust sampling were performed by the 20th week of pregnancy, immediately after birth, and then at 1 years of age. Der p 1 was assayed using a two‐site immunometric ELISA. CBMC immunoproliferative responses (AIM V serum‐free medium; 1 × 105 cells/well) were measured for 225 neonates (171 had a high risk of atopy (HR) – both parents skin test positive; 59 had a low risk of atopy (LR) – both parents skin test negative, no history of atopy) by 3H‐Thymidine (1µCi/well) incorporation after stimulation in primary culture with phytohaemagglutinin (PHA) (1 µg/mL), house dust mite [HDM] extract (30 µg/mL), immunopurified Der p 1 (30 µg/mL), Tetanus toxoid (TT) (aluma free, 30 Lf/mL) or vehicle. Blood was collected from 144 infants at the age of 1 years and stimulated proliferative responses were assessed using the same procedure.Results PHA‐stimulated lymphoproliferative response was significantly lower in HR compared to LR neonates (mean difference 38%, 95% CI 15%−54%; P = 0.003); significantly lower proportion of positive CBMC responses to HDM occurred in LR than in HR neonates (30.4% vs. 46.6%; P = 0.034). There was no relationship between Der p 1 levels in maternal bed and CBMC immunoproliferative responses, despite the 21 000‐fold range of maternal Der p 1 exposure. No significant differences in magnitude, or in proportion of positive responses to any stimulant were observed between the neonates at low, medium or high tertile of allergen exposure. Immunoproliferative responses at birth were not predictive of 1‐year PBMC responses. There was no relationship between maternal allergen exposure in pregnancy and 1‐year PBMC proliferative responses. However, the proportion of positive proliferative responses at 1 years significantly increased with increasing infant Der p 1 exposure at 1 years.Conclusion These results indicate that the magnitude of immunoproliferative responses are unrelated to maternal mite allergen exposure and cannot be used as evidence for in utero sensitization to inhalant allergens. The immunoproliferative responses at 1 year seem to shift away from the genetically influenced responses at birth towards responses to specific stimulants which correlate with environmental exposure to those specific stimulants. These data support the concept of sensitization to inhalant allergens occurring in early life, but not in utero.
Local increase in the number of mast cells and expression of nerve growth factor in the bronchus of asthmatic patients after repeated inhalation of allergen at low‐dose
Authors: Kassel O. , De Blay F. , Duvernelle C. , Olgart C. , Israel‐Biet D. , Krieger P. , Moreau L. , Muller C. , Pauli G. , Frossard N. 
Published: March 2003

Background Repeated inhalation of allergen at low‐dose induces an increase in bronchial hyper‐responsiveness, without any associated symptom. The concomitant events in the bronchus have not been described.Objective We have studied the dynamic number of mast cells in the airways of patients with mild asthma before and after repeated inhalation of allergen at low‐dose and the expression of nerve growth factor (NGF), which is reported to promote growth and survival of mast cells.Methods Twelve patients with mild asthma to cat allergen were enrolled at random in a blind placebo‐controlled study, and submitted to repeated low‐dose allergen exposure (1/5 of the provocative dose). Mast cells were immunolocalized using an antibody against mast cell tryptase. NGF and its high affinity receptor, TrkA, were immunolocalized using anti‐NGF and anti‐TrkA antibodies, respectively. NGF mRNA was quantified by competitive polymerase chain reaction (PCR) after reverse transcription of total RNA extracted from bronchial biopsy. NGF protein levels were measured by ELISA in bronchoalveolar lavage (BAL) fluid.Results Bronchial mast cell number was increased significantly after allergen exposure as compared with before. NGF expression in the bronchus was immunolocalized mainly to epithelial cells, but also to fibroblasts, blood vessels, and a few infiltrated cells. NGF mRNA levels in bronchial biopsies were increased significantly after allergen exposure. The high affinity receptor for NGF, TrkA, was immunolocalized to the infiltrated mast cell membrane.Conclusion Our study shows that the increase in the number of mast cells and in the expression of NGF induced by allergen exposure in the bronchus of asthmatic patients is occurring before the onset of symptoms. In addition, our finding of the presence of the TrkA receptor on the membrane of the infiltrated mast cell in situ brings evidence of the mast cell as a target cell for the growth factor activity of NGF in the airways in asthma.
Degranulation influences heparin‐associated inhibition of RT‐PCR in human lung mast cells
Authors: Glaum M. C. , Wang Y. , Raible D. G. , Schulman E. S. 
Published: March 2003

Background The study of gene expression from human lung mast cells (HLMC) has been limited by the ability to reliably detect mRNA transcripts from scant quantities of mast cell RNA contaminated with heparin.Objective As heparin is granule‐associated within the mast cell, we examined the role of degranulation in altering the intrinsic ability of this proteoglycan to inhibit reverse transcription polymerase chain reaction (RT‐PCR) from HLMC RNA. We also explored alternative means of RNA isolation to eliminate primary heparin contamination.Methods Purified HLMC (> 90% pure) were challenged for 2 h with buffer, anti‐IgE (3 µg/mL) and/or ionophore A23187 (100 ng/mL) or phorbol 12‐myristate 13‐acetate (50 ng/mL). Histamine release was measured using a spectroflourometric assay. Following challenge, RNA was isolated by either phenol‐chloroform extraction or by nitrocellulose spin column. Parallel samples were either treated with heparinase or placed on ice for 2 h prior to reverse transcription. PCR was performed using primers specific for the housekeeping gene, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH).Results In each of five studies, GAPDH bands were detected at greater intensity in total cellular RNA (tcRNA) derived from degranulated than from non‐degranulated mast cells. However, when examining heparinase‐treated tcRNA from the same mast cell samples, the intensity of GAPDH bands normalized between degranulated and non‐degranulated cells. When comparing parallel samples of column‐purified tcRNA, subsequent treatment of samples with heparinase had no effect on the detection of GAPDH, indicating that endogenous heparin was effectively removed by the technique. Moreover, no variability was noted in GAPDH signal detected from resting vs. degranulated mast cells (n = 3).Conclusions Degranulation influences the degree of heparin‐associated inhibition of RT‐PCR in HLMC, and that spin column purification of tcRNA is a time‐saving, effective alternative to heparinase pre‐treatment.
A polymorphism in the coding region of interleukin‐13 gene is associated with atopy but not asthma in Chinese children
Authors: Leung T. F. , Tang N. L. S. , Chan I. H. S. , Li A. M. , Ha G. , Lam C. W. K. 
Published: March 2003

Background Interleukin (IL)‐13 is an important cytokine secreted from type 2 helper T lymphocytes. It is essential for modulating IgE synthesis by human B cells. Previous studies showed that polymorphisms in the IL‐13 gene were associated with serum total IgE or allergic asthma. The relationship of this marker with sensitization to individual aeroallergens has not been evaluated.Objective We tested whether a polymorphism in the coding region of the IL‐13 gene is associated with asthma and atopy in asthmatic children in Hong Kong.Methods We used restriction fragment length polymorphism to detect R130Q genotype in Chinese children with asthma and control subjects. Serum total IgE was measured by microparticle immunoassay and specific IgE to common aeroallergens was measured using fluorescent enzyme immunoassay. Pulmonary function studies were performed using spirometry.Results One hundred and fifty‐seven patients and 54 control children were recruited. Their mean serum total IgE concentrations were 994 kIU/L and 473 kIU/L, respectively (P < 0.0001). Atopy as defined by ≥ 1 positive RAST was found in 141 patients and 32 control children. The GlnGln form of the R130Q polymorphism in the IL‐13 gene was associated with serum total IgE (P = 0.005) as well as specific IgE to Der p 1 (P = 0.021), mixed cockroaches (P = 0.03) and dog (P = 0.003) but not with physician‐diagnosed asthma (P = 0.621). In addition, the R130Q polymorphism did not correlate with subjective or objective indicators of asthma severity in our patients.Conclusion Our results suggest that the R130Q polymorphism of the IL‐13 gene is associated with elevated serum total and allergen‐specific IgE but not asthma in Chinese children.
Selective protective effects of nitric oxide inhalation on allergen‐induced acute airway reactions in the pig
Authors: Middelveld R. J. M. , Alving K. 
Published: March 2003

Background Nitric oxide (NO) is thought to be an important mediator of inflammatory processes during allergic reactions in the respiratory tract.Objective This study was undertaken to investigate the effects of inhalation of NO on the allergen‐induced acute airway reactions in the pig.Methods Specific pathogen‐free pigs were sensitized with Ascaris suum antigen and challenged with an allergen aerosol during mechanical ventilation and anaesthesia. One group (n = 8) was treated with inhaled NO (20 ppm) which was given from 30 min before allergen challenge until the experiments were completed at 120 min after challenge. A control group (n = 8) did not receive NO (< 0.001 ppm).Results Inhalation of 20 ppm NO prevented the fall in arterial pO2/FiO2 levels that was observed in the control group (areas under the curve between 0 and 120 min were 3.7 ± 1.4 kPa/min in NO‐treated pigs vs. 15.9 ± 3.4 in controls, P < 0.01, Mann–Whitney U‐test) and it decreased baseline pulmonary arterial pressure (change from time‐point − 30–0 was 3.1 ± 5.3% in the control and − 19.9 ± 3.5% in the NO group, P < 0.01), which in turn resulted in a lower pulmonary arterial pressure during allergen challenge. NO also caused vasodilatation in the bronchial circulation, resulting in increased bronchial vascular conductance throughout the experiment. NO inhalation caused a small, but non‐significant, reduction in the allergen‐induced bronchoconstrictor response, whereas histamine release, as detected in urine, was not changed. Total protein levels in bronchoalveolar lavage (BAL) fluid were significantly decreased in the NO group at 120 min after challenge compared with 45 min (373 ± 101 µg/mL vs. 631 ± 184, respectively, P < 0.05, Wilcoxon matched pairs test), whereas levels in the control group did not change between these two time‐points (513 ± 282 vs. 599 ± 354, not significant).Conclusion These findings indicate that NO inhalation improves ventilation/perfusion matching and causes some bronchodilatation during the allergen‐induced acute airway reaction, whereas histamine release is not affected. Moreover, NO inhalation enhanced the clearance of extravasated protein in the airways, possibly through increased bronchial blood flow. Even though some protective effects were seen, this study does not support a therapeutic role for exogenous NO in acute allergic reactions.
Grass pollen immunotherapy inhibits seasonal increases in basophils and eosinophils in the nasal epithelium
Authors: Wilson D. R. , Irani A.‐MA. , Walker S. M. , Jacobson M. R. , Mackay I. S. , Schwartz L. B. , Durham S. R. 
Published: March 2003

Background Symptoms of allergic rhinitis are accompanied by infiltration of the nasal mucosa with inflammatory cells, predominantly eosinophils and metachromatic cells (basophils and mast cells). Specific immunotherapy (IT) reduces mucosal eosinophilia and numbers of metachromatic cells in the epithelium. A specific marker distinguishing basophils from mast cells was recently developed.Objectives The basophil‐specific monoclonal antibody 2D7 was used to determine the influence of subcutaneous IT on numbers of nasal mucosal basophils compared with the effects of IT on neutrophils, eosinophils and mast cells.Method During a randomized, placebo‐controlled trial of grass pollen IT in 44 adults with severe summer hay fever, nasal biopsies were taken at baseline, out of the pollen season, and at the peak of the pollen season following 2 years treatment. Biopsies were processed for immunohistochemistry for basophils (2D7+), mast cells (AA1+), eosinophils (MBP+) and neutrophils (neutrophil elastase+).Results In placebo‐treated (PL) patients there were significant seasonal increases in basophils (P < 0.01), mast cells (P < 0.05) and eosinophils (P = 0.002) in the nasal submucosa. In IT‐treated patients significant increases in 2D7+ cells (P < 0.01) and eosinophils (P = 0.01) but not AA1+ cells (P = 0.9) were observed. These differences were significant between groups for eosinophils (P < 0.05). In the epithelium there were seasonal increases in AA1+ cells and eosinophils in both groups (PL: P < 0.01, IT: P < 0.05 for both). The between‐group difference was significant for eosinophils (P = 0.05). Basophils were observed in the epithelium of six out of 17 in the placebo group and one out of 20 in the IT group (P = 0.03). Neutrophil numbers remained constant in both epithelium and submucosa.Conclusion Successful grass pollen immunotherapy was associated with inhibition of seasonal increases in basophils and eosinophils, but not mast cells or neutrophils within the nasal epithelium. Immunotherapy may act, at least in part, by reducing seasonal recruitment of basophils and eosinophils into the epithelium.
Crete: does farming explain urban and rural differences in atopy?
Authors: Barnes M. , Cullinan P. , Athanasaki P. , MacNeill S. , Hole A. M. , Harris J. , Kalogeraki S. , Chatzinikolaou M. , Drakonakis N. , Bibaki‐Liakou V. , Newman Taylor A. J. , Bibakis I. 
Published: March 2003

Background Urban‐rural differences in the prevalence of atopy and associated allergic diseases have been reported in several non‐European countries. Within Europe, where such variations are less striking, a farm childhood has been postulated to be protective.Objective We aimed to compare the prevalence of atopy in urban and rural children living in Crete and to examine the role of early exposure to a farming environment in explaining any measured differences.Methods A cross‐sectional survey of children attending secondary schools in Iraklion, the capital city, and five villages 100 km to the south. Atopy was determined by the responses to skin prick tests with seven common aeroallergens.Results Nine‐nundred and ninety‐seven children participated in the survey. 19.6% of those living in Iraklion, but only 9.6% of those from the villages, had a positive response to one or more skin prick tests. Among urban children there were significant gradients in the prevalence of atopy across several categories of animal contact and consumption of farm (unpasteurized) milk products – before the age of 5 years. These differences, however, were not apparent among the rural children.