bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

BackgroundHuman 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.ObjectiveTo identify whether IL‐4 and IL‐5 are stored within mast cell secretory granules.MethodsWe 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.ResultsImmunoreactivity 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.ConclusionHuman 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

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

BackgroundStem 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.ObjectiveWe examined the survival promotion activity of nerve growth factor (NGF) on cord blood‐derived human cultured mast cells.MethodsExpression 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.ResultsThe 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.ConclusionNGF 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

BackgroundAllergy 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.ObjectiveThe 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.MethodsNinety‐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.ResultsMost 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.ConclusionPeach 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

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