bluuebunny/crossref_metadata_2025 huggingface dataset:
Sequence analysis and expression of a cDNA clone encoding a 98‐kDa allergen in Dermatophagoides farinae
Authors: Tsai L.‐C. , Sun Y.‐C. , Chao P.‐L. , Ng H.‐P. , Hung M.‐W. , Hsieh K.‐H. , Liaw S.‐H. , Chua K.‐Y.
Published: March 2003

Background

The important dust mite allergens identified to date are of molecular weights ranging from 14 to 60 kDa. Our previous protein study indicated that the 98‐kDa native paramyosin in Dermatophagoides farinae mite showed IgE reactivity with 82% of the mite‐sensitive asthmatic patients suggesting that it is a novel major mite allergen. This study described the isolation and characterization of the cDNA clone encoding the 98‐kDa mite allergen.

Methods

A Dermatophagoides farinae cDNA library was constructed in lambda ZAPII vector and the library was immunoscreened with a monoclonal antibody 642. The cDNA insert was sub‐cloned into M13 sequencing vector for single‐stranded sequencing. The whole cDNA insert was expressed in pGEX‐2T Escherichia coli expression system as a fusion protein with GST. The allergenicity of the recombinant peptides was tested by skin tests and IgE immunoassay. The IgE and IgG immunoassays were performed with sera from 20 mite‐allergic patients.

Results

The cDNA clone Df642 was 2134 bp long, coding for a polypeptide of 711 amino acid residues. Protein sequence analysis and alignment confirmed that the deduced polypeptide is a mite paramyosin which is truncated slightly at the N‐ and C‐terminuses. In vivo skin tests and in vitro IgE‐binding study showed that 62% (13/21) and 50% (10/20) of the mite‐sensitive asthmatic patients reacted positively with the recombinant Dermatophagoides farinae paramyosin, respectively.

Conclusion

The study indicated that 98‐kDa mite paramyosin is an important allergen.
Citrus red mite (Panonychus citri) is the most common sensitizing allergen of asthma and rhinitis in citrus farmers
Authors: Kim  , Son  , Kim  , Park  , Lee  , Cho  , Min  , Kim
Published: March 2003

Objective

To evaluate type I hypersensitivity to citrus red mite (Panonychus citri), its prevalence, and relationship to respiratory dysfunction, a cross‐sectional survey was performed among citrus farmers on Cheju Island, Korea.

Materials and methods

Questionnaires, and skin prick test responses to 11 common inhalant allergens and citrus red mite were performed in 181 citrus farmers, and serum‐specific IgE antibodies to citrus red mite were measured by ELISA in sera of 123 subjects. To determine airway hyperresponsiveness, methacholine bronchial provocation tests were performed in 55 subjects who complained of recurrent lower respiratory symptoms.

Results

The prevalence of asthma‐based on presence of asthmatic symptoms on the questionnaire and airway hyperresponsiveness to methacholine, and allergic rhinitis based on presence of nasal symptoms on the questionnaire and positive skin‐test response were 12.1% and 19.3%, respectively. The positive rate of skin responses to one or more of 11 common inhalant allergens excluding citrus red mite was 17.1%, and if citrus red mite was included, 25.9% of farmers had positive responses. On skin prick tests, citrus red mite (16.5%) was the most common sensitizing allergen, followed by cockroach (11.0%), Dermatophagoides pteronyssinus (9.9%), and D. farinae (9.3%). Among farmers with asthma and allergic rhinitis, the positive skin responses to citrus red mite were noted in 54.5 and 68.5%, respectively. Serum‐specific IgE antibodies to citrus red mite were detected in 45 farmers (36.5%) of the 123 tested, and there was significant correlation between specific IgE level and weal (A/H ratio) to citrus red mite (r = 0.57, P < 0.001). The prevalence of asthma was higher in subjects with positive skin responses or high serum‐specific IgE antibodies to citrus red mite than in those without skin response or serum specific IgE (P < 0.05, respectively).

Conclusion

Citrus red mite is the most important allergen in citrus farmers with asthma and rhinitis in which causative allergen has not been identified. It should be included in the skin test battery for screening the causative allergen in farmers exposed to citrus red mite.
Specific inhibition of TH2‐type cytokine production from human peripheral T cells by terfenadine in vitro
Authors: Munakata  , Umezawa  , Iwata  , Dong  , Yoshida  , Ishii  , Morimoto
Published: March 2003

Background

Cytokine imbalance is thought to be one of the causes for allergic diseases. The effect of anti‐allergic drugs on cytokine production from T cells should be examined in a convenient way.

Objectives

To study the in vitro effect of terfenadine, a prototype non‐sedating H1 receptor antagonist, on cytokine production from activated T cells.

Methods

T cells were cultured in the presence of terfenadine on anti‐CD3 mAb and anti‐CD26 mAb‐coated wells, anti‐CD3 mAb and anti‐CD28 mAb‐coated wells, and anti‐CD3 mAb wells with PMA. T‐cell proliferation, along with the concentrations of interleukin (IL) ‐2, interferon (IFN) ‐γ, IL‐4, and IL‐5 were measured.

Results

Terfenadine inhibited T‐cell proliferation and IL‐4 and IL‐5 production under each costimulatory condition tested, whereas it had no effect on IL‐2 and IFN‐γ production.

Conclusions

These results indicate that terfenadine has a specific inhibitory effect on TH2‐type cytokine production induced by several ways of costimulatory activation.
Allergy to bovine β‐lactoglobulin: specificity of human IgE to tryptic peptides
Authors: Sélo  , Clément  , Bernard  , Chatel  , Créminon  , Peltre  , Wal
Published: March 2003

Background

Bovine β‐lactoglobulin (Blg) is a major cow's milk allergen. It is the main whey protein, without any counterpart in human milk. Blg chemical hydrolysates appeared to retain most of the immunoreactivity of the native protein. Allergenicity of Blg has already been shown to be associated with the four peptides derived from cyanogen bromide cleavage of Blg.

Objectives

To map the major allergenic epitopes (e.g. regions of the molecule able to bind IgE) on Blg using specific IgE from sera of 46 milk‐allergic patients as a probe.

Methods

Direct and competitive inhibition enzyme immunoassays involving immobilized native protein or purified peptides derived from Blg tryptic cleavage.

Results

Several peptides capable of specifically binding human IgEs were identified and were classified according to the intensity and frequency of the responses. The major epitopes appeared to be fragments (41–60), (102–124) and (149–162) recognized by 92, 97 and 89% of sera, respectively, whilst a second group which contained the fragments (1–8) and (25–40) was recognized by 58 and 72% of the population. A third group, comprising peptides (9–14), (84–91) and (92–100), was still detected by more than 40% of sera.

Conclusion

Three peptides were identified as major epitopes, recognized by a large majority of human IgE antibodies. Numerous other epitopes are scattered all along the Blg sequence.
Composition and stability of allergenic extracts made from gamma‐irradiated rye grass (Lolium perenne) pollen
Authors: Cadot  , Van Hoeyveld  , Ceuppens  , Stevens
Published: March 2003

Background

Phenol is commonly added to allergenic extracts as a bacteriostatic agent, but it is poisonous and also detrimental to proteins, which accelerates extract degradation. Sterilization by gamma‐irradiation of the source material could be an alternative to the use of phenol.

Objective

To analyse the potential effects of gamma‐irradiation of pollen on the composition, potency, and stability of the resulting extract, and compare them with those of phenol.

Methods

Ryegrass (Lolium perenne) pollen was sterilized by gamma‐irradiation at a dose of 25 kGy. Extracts prepared from the irradiated pollen were then compared by electrophoresis techniques and RAST inhibition to extracts, without or with 0.5% phenol, from nonirradiated pollen. In addition, proteolytic activity was compared in extracts from irradiated and nonirradiated pollen. To evaluate the stability of extracts on storage, they were analysed after forced degradation for up to 7 days at 37°C.

Results

When fresh extracts were analysed, there were no noticeable differences between the three types, as judged by immunoblotting and RAST inhibition experiments. However, on storage, extracts from irradiated pollen appeared to be superior to extracts from nonirradiated pollen, as some proteins were more stable in the former. This could be related to the lower proteolytic activity we have also observed in extracts from irradiated pollen. In contrast, extracts containing phenol degraded much faster, as proven by all our methods of investigation.

Conclusion

Gamma‐irradiation of pollen did not influence the IgE‐binding capacity of the resulting extracts, but did yield extracts with somewhat improved stability, probably by reducing the proteolytic activity. It may be concluded that gamma‐irradiation of the source material represents a good alternative to the use of phenol for the preparation of allergenic extracts.
The metabolism of antihistamines and drug interactions: the role of cytochrome P450 enzymes
Authors: Renwick A. G.
Published: March 2003

The non‐sedating antihistamines show a diversity of fates in the body and the parent drugs and metabolites may differ in their biological properties. Clinically significant interactions with inhibitors of cytochrome P450 have been reported primarily for terfenadine, which has the potential for cardiac toxicity, and is metabolized to fexofenadine, an antihistamine without cardiac effects. Astemizole shares many of these characteristics and important safety‐related interactions are likely. Loratadine undergoes extensive metabolism so that pharmacokinetic interactions could occur, but they would be of little clinical importance because of the lack of cardiac activity of the parent drug and its metabolites. Ebastine also undergoes pharmacokinetic interactions, the significance of which is dependent on clarification of the extent of any relevant cardiotoxicity of both ebastine and its metabolite. Interactions would not be clinically important for cetirizine and fexofenadine which do not show cardiac effects and are eliminated with little metabolism.
Airborne endotoxin exposure and the development of airway antigen‐specific allergic responses
Authors: Wan  , Li  , Lin
Published: March 2003

Background and objective

Repeated exposure of aerosolized antigen via respiratory tract can induce immunoglobulin (Ig) E isotype‐specific tolerance to this antigen. However, the atopic individuals often produce a higher titre of IgE in response to airborne environmental allergens. The mechanisms of this differential regulation of airway allergen‐specific immune responses are not fully understood. This study investigated the role of airborne endotoxin on the initiation of antigen‐specific airway allergic responses.

Methods

ELISA methods for detection of isotypes of antigen‐specific antibodies and competitive reverse transcription polymerase chain reaction for detection mRNA of cytokines were used. In addition, Liu stain method was used to analyse the amounts of eosinophils in bronchoalveolar lavage fluid.

Results

Mice pre‐exposed with airborne endotoxin mounted significantly higher amounts of OVA‐specific IgE antibody responses to inhaled OVA than those OVA‐only sensitized mice. Inhaled endotoxin could downregulate repeated airway antigen exposure‐induced IgE isotype‐specific tolerance and increase antigen‐induced lung eosinophils infiltration.

Conclusions

These data show that airborne endotoxin exposure could potentiate allergen‐specific airway inflammation. The results should have potential implications for understanding the development of allergen‐induced airway allergic responses.
Localization of interleukin (IL) ‐4 but not IL‐5 to human mast cell secretory granules by immunoelectron microscopy
Authors: Wilson  , Shute  , Holgate  , Howarth  , Bradding
Published: March 2003

Background

Human mast cells synthesize and secrete many cytokines of relevance to the pathogenesis of allergic diseases such as asthma and rhinitis. In particular, interleukin (IL) ‐4 and IL‐5 are likely to play key roles in the development of the inflammatory response that characterizes these diseases. Immunohistochemical studies on human nasal and bronchial mucosal biopsies suggest that IL‐4 and IL‐5 may be stored preformed in mast cells.

Objective

To identify whether IL‐4 and IL‐5 are stored within mast cell secretory granules.

Methods

We used immunogold electron microscopic analysis on bronchial mucosa and lung parenchyma from resected lung specimens, and a nasal mucosal biopsy from a patient with active allergic rhinitis. Samples were fixed in 4% paraformaldehyde plus 0.5% glutaraldehyde and processed into Lowicryl K4M resin by the ‘Progressive Lowering of Temperature’ technique. Ultrathin sections were stained immunohistochemically by an indirect immunogold method.

Results

Immunoreactivity for IL‐4, but not IL‐5, was localized to the granules of mast cells in all tissue samples. IL‐5 was localized to the matrix of eosinophil granules in these samples, but neither cytokine was detected in T cells. IL‐4 immunoreactivity increased in the granules of mast cells 24 h after immunoglobulin (Ig) E‐dependent activation (mean 17.5 ± 1.4 gold particles per granule) compared with nonactivated mast cells (mean 6.8 ± 0.8 gold particles per granule, P < 0.001), suggesting replenishment of stores by newly generated protein. Immunoreactive IL‐5 remained undetectable in mast cells 24 h after activation, a time point at which they are known to secrete large quantities of this cytokine.

Conclusion

Human mast cells store IL‐4 within the matrix of their granules. Very few, if any, lung or nasal mast cells store IL‐5. A store of preformed IL‐4 within mast cell granules is likely to have an important influence during the initiation and maintenance of the allergic immunological response.
N‐terminal sequences of high molecular weight allergens from celery tuber
Authors: Ganglberger  , Radauer  , Grimm  , Hoffmann‐Sommergruber  , Breiteneder  , Scheiner  , Jensen‐Jarolim
Published: March 2003

Background

Celery tuber is an important source of food allergens. Low molecular weight celery allergens were identified as homologues of Bet v 1 and profilin. Little is known about the relevant allergens with molecular weights between 45 and 60 kDa, which cross‐react with other plant food and pollen allergens.

Objective

The aim of this study was to isolate cross‐reactive, high molecular weight allergens from celery and to characterize them by N‐terminal sequencing.

Methods

High molecular weight allergens of celery were identified by immunoglobulin (Ig) E immunoblotting with patients' sera, and the IgE‐binding patterns were compared with those of the monoclonal antibirch pollen antibody BIP3, as well as of a polyclonal rabbit anti‐Art v 1 antiserum. Two independent methods, elution from preparative SDS‐PAGE or anion exchange chromatography, were used to purify the IgE‐binding celery proteins of interest. The isolated proteins were examined by N‐terminal sequencing and IgE‐immunoblots.

Results

Celery allergens with molecular masses of 55, 58 and 63 kDa, which were also recognized by the monoclonal BIP3 antibody and a polyclonal anti‐Art v 1 antiserum, were isolated. The 63‐kDa allergen was N‐terminally blocked. The 55‐ and 58‐kDa compounds yielded the same N‐terminus, which showed no homology to known proteins in the databases.

Conclusion

The combination of two independent protein separation techniques, immunoblotting and N‐terminal sequencing, identified an N‐terminus of two allergens in the 60‐kDa molecular weight region. Our data will be helpful for the definite molecular characterization of these important cross‐reactive molecules.
Nerve growth factor prevents apoptosis of cord blood‐derived human cultured mast cells synergistically with stem cell factor
Authors: Kanbe  , Kurosawa  , Miyachi  , Kanbe  , Saitoh  , Matsuda
Published: March 2003

Background

Stem cell factor (SCF) has been identified as a critical survival factor of human mast cells. Other cytokines which possess survival promotion activity on human mast cells are less known.

Objective

We examined the survival promotion activity of nerve growth factor (NGF) on cord blood‐derived human cultured mast cells.

Methods

Expression and function of NGF receptors on the mast cells were examined by RT PCR, flowcytometric analysis, immunoprecipitaion and western blotting. The survival promotion activity of NGF to the mast cells was examined. To evaluate the proliferating activity of NGF on the human cultured mast cells, flow cytometric analysis with propidium iodide staining was applied. To confirm whether the human mast cell growth activity of NGF was caused by a suppression of apoptosis, the proportion of the cells containing in situ DNA fragmentation was counted.

Results

The human cultured mast cells expressed the high affinity receptor p140trk but not the low affinity receptor p75LNGFR. NGF induced the phosphorylation of p140trk. NGF alone could not support the survival of the mast cells, however, the addition of NGF to the culture medium containing recombinant SCF led to a significant increase of the number of survival mast cells. No significant changes of the cell cycle from G0/G1 phase to the S/G2 + M phases were observed by NGF. In contrast, the addition of NGF to the medium with SCF showed a significant inhibitory effect on the apoptosis of the mast cells.

Conclusion

NGF may act as a key factor to promote the survival of human mast cells synergistically with SCF through the prevention of apoptosis.
Allergy to plant‐derived fresh foods in a birch‐ and ragweed‐free area
Authors: Cuesta‐Herranz J. , Lázaro M. , Figueredo E. , Igea J. M. , Umpiérrez A. , De‐Las‐Heras M.
Published: March 2003

Background

Allergy to plant‐derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross‐reactivity to pollens.

Objective

The aim of this study has been to evaluate allergy to plant‐derived fresh foods among pollen‐allergic patients from a birch and ragweed‐free area.

Methods

Ninety‐five pollen‐allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods.

Results

Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen‐allergic patients had positive skin test responses to some plant‐derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21.05% of the pollen‐allergic patients were allergic to some type of plant‐derived fresh food. Peach was the plant‐derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing.

Conclusion

Peach was the most important allergy provoking fruit in a birch and ragweed free‐area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.
Rapid polarization of Th2 cells during induction of antigen‐specific IgE antibodies in vitro
Authors: Åkesson  , Ingvarsson  , Brady  , Moynagh  , Borrebaeck
Published: March 2003

Background

Type 2 T‐helper cells (Th2) are involved in the regulation of the humoral immune response against antigens and allergens and directly affect which isotype will be produced. The mechanism that regulates antigen‐specific IgE secretion and immune deviation is still not known.

Objectives

To delineate mechanisms behind antigen‐specific IgE secretion we have used in vitro immunization and focused on T‐cell phenotype and the activation status of the transcription factor NFκB.

Methods

Peripheral blood lymphocytes (PBMC) from seronegative donors were immunized in vitro with a peptide consisting of both a T‐cell and a B‐cell epitope.

Results

Antigen‐specific IgE antibodies could be detected after a primary immunization, during which T‐helper cells secreted type 2 cytokines. Specific IgE was also detected in the secondary immunization, but due to a rapid polarization from Th2 to Th1 phenotype, exogenous IL‐4 was required for the specific IgE secretion. Analysis of NFκB activation in B and T cells during primary and secondary immunization showed that NFκB could be detected in both B and T cells during primary immunization, but was dependent on exogenous IL‐4 in the secondary immunization.

Conclusion

This is the first evidence of antigen‐specific IgE induction in vitro using naive B cells, demonstrating the involvement of T‐helper cell phenotype and NFκB and demonstrates the usefulness of in vitro cultures to study the effect of antigens on human immunocytes.
Characterization of increased cough sensitivity after antigen challenge in guinea pigs
Authors: Liu Q , Fujimura M. , Tachibana H. , Myou S. , Kasahara K. , Yasui M.
Published: March 2003

Increased sensitivity of cough reflex is a fundamental feature of bronchodilator resistant non‐productive cough associated with eosinophilic tracheobronchitis. Our hypothesis is that cough sensitivity is increased by airway allergic reaction characterized by airway eosinophilic inflammation. The aim of this study was to elucidate the hypothesis and clarify the characteristics of the increased cough sensitivity.

Number of coughs elicited by inhalation of increasing concentrations of capsaicin (10−8, 10−6 and 10−4 m) was counted 24 h after an aerosolized antigen or saline in actively sensitized or non‐sensitized (naive) conscious guinea pigs and then bronchoalveolar lavage was performed. The cough response was also measured 1 day before and 1, 2, 3, 5 and 7 days after an aerosolized antigen challenge in sensitized or naive animals. In addition, effect of procaterol (0.1 mg/kg), atropine (1 or 10 mg/kg), phosphoramidon (2.5 mg/kg) given intraperitoneally 30 min before the capsaicin challenge or capsaicin desensitization on the cough response was examined. Furthermore, the thromboxane A2 (TXA2) receptor antagonist S‐1452 in a dose of 0.01 or 0.1 mg/kg or vehicle (saline) was given intraperitoneally at 24 and 1 h before the measurement of cough response.

Number of coughs caused by capsaicin was extremely increased 24 h after an antigen challenge in sensitized guinea pigs compared with a saline or an antigen challenge in naive animals or a saline challenge in sensitized animals. The increased cough response disappeared at 3–7 days after the antigen challenge. Eosinophils in bronchoalveolar lavage fluid obtained after the measurement of capsaicin‐induced coughs, which was performed 24 h after the antigen challenge, were significantly increased in sensitized guinea pigs. The eosinophil count was significantly correlated to the number of capsaicin‐induced coughs. Procaterol or atropine did not alter the antigen‐induced increase of cough sensitivity, whereas atropine did reduce the cough response in naive animals. Phosphoramidon increased the number of capsaicin‐induced coughs in naive guinea pigs but not in sensitized and antigen‐challenged animals. Capsaicin desensitization decreased the cough response in both antigen‐challenged sensitized guinea pigs and naive animals. S‐1452 reduced the antigen‐induced increase of cough response in sensitized guinea pigs, but not in naive animals.

Airway allergy accompanied with airway eosinophilia induces transient increase in cough sensitivity, which is not mediated by bronchoconstriction. The increased cough sensitivity may result in part from inactivation of neutral endopeptidase and TXA2, one of the inflammatory mediators.
Infection of mice with the helminth Strongyloides stercoralis suppresses pulmonary allergic responses to ovalbumin
Authors: Wang Chun‐Chi , Nolan T. J. , Schad G. A. , Abraham D.
Published: March 2003

Asthma and helminth infections induce similar immune responses characterized by the presence of peripheral blood eosinophilia and elevated serum IgE levels. Epidemiological surveys have reported either increases or decreases in the development of atopic diseases and asthma based on the prevalence of helminth infections in the population.

The aim of this study was to determine if a pre‐existing helminth infection would increase or decrease subsequent allergic responses to an unrelated allergen in the lungs.

BALB/cByJ mice were infected with the nematode parasite Strongyloides stercoralis prior to ovalbumin (OVA) immunization and intratracheal challenge. Bronchoalveolar lavage (BAL) and fluid (BALF) were collected 3 days post‐challenge and cellular and humoral immune responses were measured.

Intracellular cytokine staining revealed increased IL‐4 and IL‐5 producing cells in BAL from mice infected with S. stercoralis before OVA sensitization. Increased IL‐5 protein levels and decreased IFN‐γ protein levels were also observed in the BALF. There was, however, no increase in airway eosinophil accumulation in mice infectd with parasites before sensitization with OVA as compared to mice exposed to OVA alone. Furthermore, eotaxin levels in the lungs induced by OVA was suppressed in mice infected with the parasite before OVA sensitization. The development of OVA specific IgE responses in BALF was also impaired in mice infected with the parasite before sensitization with OVA.

These results suggest that a pre‐existing helminth infection may potentiate a systemic Type 2‐type response yet simultaneously suppress in the lungs allergen‐specific IgE responses and eotaxin levels in response to subsequent exposure to allergens.
Diet and chronic obstructive pulmonary disease: independent beneficial effects of fruits, whole grains, and alcohol (the MORGEN study)
Authors: Tabak C. , Smit H. A. , Heederik D. , Ocké M. C. , Kromhout D.
Published: March 2003

Background In recent years antioxidants, foods rich in antioxidants (e.g. fruits, vegetables) and fish have been suggested to protect against chronic obstructive pulmonary disease (COPD). There are also indications for a protective effect of whole grain intake and of consuming moderate amounts of alcohol. It is, however, not clear whether the effects of the different dietary factors on COPD are independent of each other and if so, whether their effects are additive.

Objective To gain more insight into the potential protective effect of diet on COPD, we studied fruit, vegetable, fish, alcohol and whole grain consumption simultaneously in relation to pulmonary function and COPD symptoms.

Methods Analysed were cross‐sectional data collected in 13 651 men and women aged 20–59 years participating between 1994 and 1997 in the MORGEN study (monitoring project on risk factors and health in The Netherlands). Regression models were adjusted for age, gender, height (for pulmonary function only), smoking, BMI and energy intake.

Results Fruit and whole grain intake showed independent beneficial associations with COPD (P‐trend < 0.001). Furthermore, in subjects with low alcohol consumption (1–30 g/day) the forced expiratory volume in 1 s (FEV1) was higher and the prevalence of COPD symptoms lower than in non‐drinkers (P < 0.001). The effects of a favourable intake of fruits (> 180 g/day), whole grains (> 45 g/day) and alcohol (1–30 g/day) were largely additive. In the 2998 subjects with a favourable intake of the three foods, the FEV1 was 139 mL higher and the prevalence of COPD symptoms lower (odds ratio (OR) = 0.44) than in subjects (n = 1406) with unfavourable intakes of fruits, whole grains and alcohol (P < 0.001). A similar effect was observed in those who had never smoked. Fish and vegetable intake did not show independent beneficial associations with COPD.

Conclusions Our results suggest independent beneficial effects of fruits, whole grains and alcohol on COPD that are largely additive and cannot be explained by smoking habits.
Identification of mutations in the genes for the pollen allergens of eastern red cedar (Juniperus virginiana)
Authors: Midoro‐Horiuti T. , Goldblum R. M. , Brooks E. G.
Published: March 2003

Background Cedar pollens are important causes of seasonal allergic disease in diverse geographical areas. However, pollens from different families and species vary in their propensity to induce allergic responses.

Objective To compare the structure of potential allergens from eastern red cedar (Juniperus virginiana) pollen with those of the highly allergenic cedar (mountain cedar, J. ashei) pollens.

Materials and methods The cDNAs for potential pollen allergens, Jun v 1 and Jun v 3, were amplified by reverse transcriptase‐polymerase chain reaction, cloned and sequenced. Expression of the native proteins in pollen was characterized by SDS–PAGE and immunoblotting.

Results The cDNA sequence for one potential major allergen, Jun v 1, was highly homologous to those of the other cedar pollens. The second potential allergen, Jun v 3, was also highly homologous to its counterpart in mountain cedar, but a stop codon in the mRNA would result in a protein of only 91 amino acids, which would lack potential N‐glycosylation sites and the IgE binding epitopes of the 199 amino acid homologue from mountain cedar pollen, Jun a 3. IgE from the sera of patients with hypersensitivity to cedar pollen bound to eastern red cedar proteins of four different sizes. N‐terminal amino acid sequence analysis indicated that two of these proteins (43 and 30 kDa) were either isoforms or processed Jun v 1. No Jun v 3 protein was detected. The N‐terminal sequence of an additional 145‐kDa allergen, termed Jun v 4, was not homologous to any previously described allergens.

Conclusion These findings suggest that mutations in the genes or post‐translational modifications of two potentially allergenic proteins might help to explain why the pollen of eastern red cedar is reported to be less allergenic than those of other members of Cupressaceae and Taxodiaceae families.
In vitro and in vivo responses to the recombinant bovine dander allergen Bos d 2 and its fragments
Authors: Ruoppi P. , Virtanen T. , Zeiler T. , Rytkönen‐Nissinen M. , Rautiainen J. , Nuutinen J. , Taivainen A.
Published: March 2003

Background About one in every four cases of occupational rhinitis recorded in Finland is animal‐induced. Bovine allergens are the most important in this respect and the largest patient group consists of dairy farmers. Allergen immunotherapy, if proven effective, safe and feasible, would be ideal for their treatment. The development of recombinant allergens has offered new potential therapeutic prospects. Fragments of recombinant Bos domesticus (Bos d 2) allergen could be suitable for this purpose because they are recognized by T cells but their IgE‐binding capacity is attenuated.

Objective The aim of this study was to verify whether the potential of the two fragments of recombinant Bos d 2 (corresponding to amino acids 1–131 and 81–172) to induce immediate allergic reaction in a shock organ (nose) was decreased compared to the complete recombinant allergen, which would be an advantageous property for a preparation intended for allergen immunotherapy.

Methods The study group consisted of 10 dairy farmers with cow‐induced allergic rhinitis. We used the IgE titres against native Bos d 2 measured by indirect IgE ELISA to characterize the level of sensitization and compared the IgE titres in the rhinitis patients with 12 cow‐sensitized asthmatic farmers and 12 healthy students. In vitro reactivity against recombinant Bos d 2 and its two fragments was studied by indirect IgE ELISA and in vivo reactivity by nasal provocation tests.

Results The IgE titres against native Bos d 2 of patients with rhinitis tended to be lower than the titres of asthmatics. The healthy students did not exhibit any detectable IgE reactivity to native Bos d 2. In the patients with rhinitis, there was no statistically significant difference between IgE responses against native and recombinant Bos d 2, whereas with both in vitro and in vivo, the reactivity to both fragments of recombinant Bos d 2 was lower than the reactivity to the complete recombinant allergen.

Conclusions Due to the decreased in vivo capacity to induce immediate allergic reactions, the fragments may be better tolerated in allergen immunotherapy than the complete allergen.
Serum albumins are the major site for in vivo formation of hapten‐carrier protein adducts in plasma from humans and guinea‐pigs exposed to type‐1 allergy inducing hexahydrophthalic anhydride
Authors: Johannesson G. , Rosqvist S. , Lindh C. H. , Welinder H. , Jönsson B. A. G.
Published: March 2003

Background Organic acid anhydrides (OAAs) are highly allergenic compounds used in the chemical industry. The OAAs probably act as haptens but the proteins that form conjugates with OAAs in vivo are still unknown. Conjugates between the anhydrides and serum albumins (SAs) have routinely been used when testing for OAA‐specific antibodies. However, the use of SA as the carrier‐protein in these tests has never been evaluated.

Objective The aim of this study was to identify major and also immunologically relevant protein conjugates of a particularly sensitizing OAA, hexahydrophthalic anhydride (HHPA), in plasma.

Methods Plasma was obtained from a HHPA‐exposed worker, from a guinea‐pig (GP) exposed to HHPA in an exposure chamber for 2 weeks (8 h/day, 5 days/week) and from a GP exposed once, nose‐only, to tritium‐labelled HHPA for 8 h. The plasma was fractionated using ion exchange chromatography and gel filtration. These fractions and also aliquots of unfractioned plasma were hydrolysed, derivatized and analysed for anhydride adduct content using gas chromatography–mass spectrometry. Further, plasma from the tritium labelled HHPA‐exposed GP was separated by SDS gel electrophoresis and analysed by autoradiography. In addition, immunologically relevant proteins were identified through specific IgE and IgG immunoblottings using sera from exposed workers.

Results For humans > 85% and for GPs > 74% of the HHPA‐adducts coeluted with SA in plasma. Autoradiography of GP‐plasma shows a single 66 kDa protein that binds HHPA. IgE immunoblotting shows a major 66 kDa and a minor 28 kDa protein which could be inhibited by HHPA‐SA conjugate. IgG immunoblotting showed a major 66 kDa protein and several minor protein bands.

Conclusion This study shows SA to be the major protein in plasma that forms adducts in vivo with HHPA. The results also show that in an in vitro synthesized HHPA plasma protein conjugate, HHPA‐specific IgE and IgG antibodies bind preferably to the SA.
Inhibition of cytokine generation and mediator release by human basophils treated with desloratadine
Authors: Schroeder J. T. , Schleimer R. P. , Lichtenstein L. M. , Kreutner W.
Published: March 2003

Background Desloratadine is a non‐sedating, clinically effective, anti‐allergic therapy that has been shown to exhibit anti‐inflammatory properties that extend beyond its ability to antagonize histamine at H1‐receptor sites. This latter effect has been shown in vitro to be both IgE‐dependent and ‐independent.

Objective In this study, we addressed the ability of desloratadine to inhibit the in vitro generation of interleukin (IL)‐4 and IL‐13 from human basophils while concurrently comparing its efficacy in preventing mediator release by these cells.

Methods Basophil‐enriched suspensions were treated with various concentrations of desloratadine for 15 min before stimulating with either anti‐IgE antibody, calcium ionophore, IL‐3 or phorbol ester. Histamine (fluorimetry), LTC4 (RIA) and IL‐4 (ELISA) were all assayed using the same 4‐h culture supernatants. IL‐13 (ELISA) was measured in supernatants harvested after 20 h incubation. IL‐4 mRNA expression (dilutional RT‐PCR) was also examined.

Results Desloratadine was found to be nearly six–seven times more potent in preventing the secretion of IL‐4 and IL‐13 induced by anti‐IgE than it was at inhibiting the release of histamine and LTC4. These cytokines were equally inhibited by desloratadine following activation with ionomycin despite the lack of an effect on the histamine induced with ionomycin. Desloratadine had a lesser effect regarding inhibition of the IL‐13 secreted in response to IL‐3 and PMA. There was no evidence that desloratadine mediated its inhibitory effects by causing decreased cell viability. Finally, IL‐4 mRNA accumulation was remarkably inhibited, by as much as 80%, following pretreatment with desloratadine.

Conclusion While capable of inhibiting histamine and LTC4 release by human basophils, desloratadine is more effective at targeting the signals regulating IL‐4 and IL‐13 generation in these cells. This inhibitory effect on cytokine generation provides additional evidence that this antihistamine exerts anti‐inflammatory properties.
Difference in exposure to airborne major rat allergen (Rat n 1) and to endotoxin in rat quarters according to tasks
Authors: Lieutier‐Colas F. , Meyer P. , Larsson P. , Malmberg P. , Frossard N. , Pauli G. , De Blay F.
Published: March 2003

Endotoxins found in occupational settings constitute a risk factor in the severity of respiratory allergic symptoms.

Objectives To assess the airborne concentrations of major rat allergen (Rat n 1) and endotoxin under various circumstances.

Methods We took 483 airborne samples from 12 sites: 114 individual samples for endotoxin measurements and 113 for Rat n 1, from 38 workers (nine animal technicians, nine laboratory technicians, nine scientists and 11 students); and 256 static samples in rat rooms and experimental rooms, with or without disturbance, for simultaneous endotoxin and Rat n 1 measurements. Rat n 1 was measured with a two‐site monoclonal ELISA and endotoxins with the Limulus method.

Results Airborne Rat n 1 and endotoxin were significantly higher in rat rooms than in experimental rooms. Animal technicians had the greatest exposure to both Rat n 1 and endotoxin. Cage cleaning and rat feeding induced the highest exposure to Rat n 1 and endotoxin. Furthermore, we observed no significant difference in endotoxin exposure between researchers with or without rat contact during the sample period. There was no correlation between the number of rats present and airborne endotoxin concentrations.

Conclusions Exposure to airborne Rat n 1 and endotoxin is higher during cleaning and feeding tasks than during any other task, we feel that a major source of both is contaminated bedding that becomes airborne during disturbance.
DNA sequence variants of the platelet‐derived growth factor A‐chain gene
Authors: Baron R. , Silverman E. S. , Grasemann H. , Collins T. , Drazen J. M.
Published: March 2003

Background Platelet‐derived growth factor A‐chain (PDGF‐A) is a potent connective tissue mitogen implicated in lung growth and development. PDGF‐A may have a role in asthma through effects on fibroblasts and bronchial smooth muscle cells.

Objective To test the hypothesis that there exist variations in the PDGF‐A gene associated with the asthma phenotype.

Methods We screened genomic DNA from normal and asthmatic subjects using single‐stranded conformational polymorphism (SSCP) for mutations in the promoter and all seven exons of the gene.

Results Four transition polymorphisms (three novel) were identified: one each in exons 3 and 4 (overall population allele frequencies 0.18 and 0.02, respectively) which did not alter the protein sequence, one in exon 4 (frequency 0.005) which resulted in a valine to isoleucine substitution, and one in intron 5 (frequency 0.5). The intron 5‐sequence variant is close to the 3′ end of exon 5 but does not appear to affect alternative splicing of PDGF‐A exon 6 RNA. The frequencies of the polymorphisms in exons 3 and intron 5 did not differ between the asthmatic and non‐asthmatic subjects, but there was a significant frequency difference between Caucasian and African‐American subjects for each of these polymorphisms (P = 0.03 and 0.003, respectively).

Conclusion No association was found between the sequence variants in the PDGF‐A gene and the development of asthma. However, the allele frequency of some of the sequence variants differed between the Caucasian and African‐American subjects.
In vivo kinetics of the immunoglobulin E response to allergen: bystander effect of coimmunization and relationship with anaphylaxis
Authors: Von Garnier C. , Astori M. , Kettner A. , Dufour N. , Corradin G. , Spertini F.
Published: March 2003

Background Murine models of hypersensitivity to allergens are useful tools for the evaluation of preclinical strategies to down‐regulate the IgE response.

Objective To monitor the long‐term kinetics of T and B cell responses to allergen as a function of allergen dosage and to investigate the effect of parallel immunization with a second antigen; to correlate B cell response with anaphylaxis.

Methods CBA/J mice were sensitized every other week by subcutaneous injections of phospholipase A2 (PLA2) and/or ovalbumin (OVA) adsorbed to alum. Specific antibody isotype responses, T cell proliferation, T cell cytokine production and anaphylaxis were assessed throughout the sensitization phase.

Results Low‐dose immunization with PLA2 (0.1 µg) favoured a long‐term, specific T helper (Th)2 response with high IgE and IL‐4 production in contrast to high‐dose PLA2 (10 µg) immunization, which biased the immune response towards a Th1 response with high IgG2a and low IL‐4 production. Parallel immunization with an unrelated antigen (ovalbumin) had a significant bystander effect on the immunization with PLA2, which was also dose‐dependent. Finally, although anaphylaxis as measured by rectal temperature drop was allergen‐specific, it could be induced in the high‐ and low‐dose immunization groups, and was not solely dependent on IgE levels.

Conclusion Though low‐dose allergen immunization appears to induce an efficient IgE response, the intensity and quality of this response may be modulated by bystander effects of parallel immunization and does not correlate strictly with anaphylaxis. This observation has relevance to the design of clinical immunotherapy protocols using murine model‐based data.
What is the role of the hevein‐like domain of fruit class I chitinases in their allergenic capacity?
Authors: Dìaz‐Perales A. , Sánchez‐Monge R. , Blanco C. , Lombardero M. , Carillo T. , Salcedo G.
Published: March 2003

Background Class I chitinases are the major panallergens in fruits associated with the latex–fruit syndrome. These enzymes contain an N‐terminal hevein‐like domain homologous to latex hevein, and a larger catalytic domain. The role of these domains in their allergenic capacity is still controversial.

Objective We sought to evaluate the role of both domains of class I chitinases in their IgE‐binding properties, using Cas s 5, the major allergen from chestnut, as a model.

Methods Recombinant Cas s 5 and its deleted form, lacking the hevein‐like domain, designated rCat, were expressed in Pichia pastoris using the pPIC 9 vector. Both recombinant products were purified from the supernatants of transformed yeast cultures by gel‐filtration and cation‐exchange chromatography. The isolated proteins were characterized by N‐terminal sequencing, enzymatic activity and N‐glycosylation tests, anti‐chitinase and specific IgE immunodetection. Immunoblot, RAST and CAP inhibition assays were also performed.

Results Both purified rCas s 5 and rCat showed the expected N‐terminal amino acid sequences and an enzymatic activity similar to that of their natural counterparts isolated from chestnut seeds, and were strongly recognized by anti‐chitinase antibodies. In contrast, only rCas s 5, but not rCat, bound specific IgE from sera of patients suffering from the latex–fruit syndrome, and fully inhibited IgE‐binding to natural Cas s 5 in immunoblot inhibition assays. Latex hevein also exerted a strong immunoblot inhibition of IgE‐binding to chestnut Cas s 5. RAST and CAP inhibition using whole chestnut extract on the solid phase, rendered inhibition levels around 70–90% for rCas s 5 and 60% for rCat, in contrast to the immunoblotting results.

Conclusions Recombinant Cas s 5 behaves like natural Cas s 5 in IgE‐binding assays in vitro. The hevein‐like domain of allergenic class I chitinases seems to include all their main IgE‐binding epitopes when tested by immunodetection and immunoblot inhibition experiments. RAST and CAP inhibition assays, on the contrary, suggest that relevant epitopes are also harboured in the catalytic domain of these allergens.
Increase of allergen‐specific immunoglobulin E antibodies from 1973 to 1994 in a Finnish population and a possible relationship to Helicobacter pylori infections1
Authors: Kosunen T. U. , Höök‐Nikanne J. , Salomaa A. , Sarna S. , Aromaa A. , Haahtela T.
Published: March 2003

Background The prevalence of atopic diseases – hayfever, asthma and eczema – has increased over the past decades. The increase may be associated with decreased rates of infections such as measles, hepatitis A, tuberculosis, toxoplasmosis, and, as recently suggested, Helicobacter pylori gastritis.

Objective Since the increase of atopy has been mainly based on clinical studies, we wanted to study the prevalence of allergen‐specific Immunoglobulin (Ig)E antibodies in two cross‐sectional, adult population‐based serum samples two decades apart. Since the sera had been tested for H. pylori antibodies, we also had a chance to look for a possible relationship between these two findings.

Methods We determined the prevalence rate of allergen‐specific serum IgE antibodies against birch and timothy pollen, and cat and dog epithelium allergens by the radioallergosorbent test in a 15–54‐years‐old Finnish population using 326 sera collected in 1973 and 319 sera collected in 1994 from randomly selected subjects.

Results From 1973 to 1994 allergen‐specific IgE prevalence rates and IgE antibody levels rose. In 1994, the prevalence rate of positive findings in 15–24‐year‐old population had increased from 11 to 38% (3.5‐fold increase, P = 0.0001, OR 5.12, CI 95% 2.32–11.3). In older 10‐year age groups similar trends did not reach significance, but the overall change was significant with all three cut‐off levels of allergen‐specific IgE analysed.

The percentage of IgE‐positive persons rose mainly in the subgroup with no H. pylori antibodies. In 1994 21% of the H. pylori‐negative subjects had IgE antibodies compared with 5% of the H. pylori‐positive subjects (in 1973 11% in both subgroups).

Conclusions IgE‐based evidence for an increase in IgE‐mediated allergy was uncovered. The increase occurred mainly in the subgroup with no antibodies to H. pylori, which support the hypothesis that H. pylori could be one of the microbes counteracting atopy.