bluuebunny/crossref_metadata_2025 huggingface dataset:
Cross–Reactivity between Dermatophagoides siboney and Other Domestic Mites
Authors: Ferrándiz R. , Casas R. , Dreborg S.
Published: April 2003

Background: Dermatophagoides siboney (Ds) cross–reacted with other mite allergens in mite–allergic patients. The aim of this study was to investigate the cross–reactivity between individual allergens responsible for this cross–reactivity. Methods: The inhibition of IgE binding to Ds allergens was investigated by non–reducing SDS–PAGE and Western blotting. Nitrocellulose membranes were incubated with a pool of sera from mite–sensitive asthmatics, after the addition of serial dilutions of D. farinae (Df), D. pteronyssinus (Dp), D. microceras (Dm), Lepidoglyphus destructor (Ld), Tyrophagus putrescentiae (Tp), Acarus siro (As) and Blomia tropicalis (Bt). N–terminal amino acid sequences and amino acid analysis of the purified major allergens, Der s 1, 2 and 3, were performed after transfer to polyvinylidene difluoride membranes. Results: The inhibition was higher with Df (86%), Dp (54%) and Dm (49%) extracts than with Ld (20%), Tp (11%), As (18%) and Bt (6%). The dose–response inhibition showed a diverse pattern for the individual allergens. Despite the high cross–reactivity between the pyroglyphid mites, some proteins of Ds were less inhibited, e.g. by the Df and Dp 80–kD protein, and by the Dm and Dp 52–, 37–, 30– and 14–kD allergens. The 65–, 62–, 37– and 30–kD proteins were always inhibited more than 50% by all the mite extracts at the maximum concentration used. The 80–, 52–, 43–, 27– and 14–kD proteins cross–reacted to a lesser extent. Individual allergens of Ds were much less inhibited by non–pyroglyphid mites. However, at the highest concentration, Ld also inhibited most of the Ds allergens. All the ten selected allergens were inhibited to some extent by the heterologous mite extracts. The N–terminal sequences of Der s 1, 2 and 3 allergens showed higher homology to Df and Dm than to Dp. The homology of the group 2 allergens was higher than that of the group 1 allergens. Conclusion: The individual allergens of Ds were more similar to Df and Dm than to Dp. There was a limited and variable cross–reactivity with non–pyroglyphid mites. No single allergen was unique for Ds.
Neopterin–Induced Tumor Necrosis Factor–· Synthesis in Vascular Smooth Muscle Cells in vitro
Authors: Hoffmann G. , Frede S. , Kenn S. , Smolny M. , Wachter H. , Fuchs D. , Grote J. , Rieder J. , Schobersberger W.
Published: April 2003

Synthesis and secretion of proinflammatory mediators like tumor necrosis factor–α and neopterin are common events in severe systemic inflammatory disorders, e.g. sepsis and septic shock. Recent data suggest that both substances show similarities with respect to their bioactivities. In the present study we investigated the potential interactions of neopterin and tumor necrosis factor–α on inducible nitric oxide synthase gene expression and nitric oxide generation in rat vascular smooth muscle cells. In addition, we studied the influence of neopterin on tumor necrosis factor–α synthesis in this cell type. Single stimulation of smooth muscle cells with either neopterin or tumor necrosis factor–α caused inducible nitric oxide synthase gene expression and nitric oxide production. Coincubation of cells with both compounds resulted in at least additive effects on nitric oxide synthesis. Quantification of tumor necrosis factor–α cDNA revealed a dose–dependent effect of neopterin on tumor necrosis factor–α gene expression. Similar results were obtained concerning the detection of tumor necrosis factor–α protein and the assessment of tumor necrosis factor–α bioactivity. These data suggest that neopterin and tumor necrosis factor–α are closely associated with regard to synthesis and effects, respectively. The interactions of both inflammatory mediators in vascular smooth muscle cells might contribute to the excessive release of nitric oxide observed during sepsis, thus triggering cellular destruction and multiple organ failure.
Prevalence of Helicobacter pylori–Associated Gastritis in Chronic Urticaria
Authors: Wedi Bettina , Wagner Siegfried , Werfel Thomas , Manns Michael Peter , Kapp Alexander
Published: April 2003

Background: Chronic urticaria and concurrent angioedema are frustrating problems for both physicians and patients. Methods: 100 patients with chronic urticaria (mean duration 33.3±48.2 months) attending the urticaria consulting hour of our Department of Dermatology within 1 year were carefully analyzed for pathogenesis to avoid extensive unnecessary diagnostic approach in the future. Results: In 43 cases a potential infectious trigger could be identified, 35 were of idiopathic origin, and 15 demonstrated pseudoallergic reactions to acetylsalicylic acid or food additives, 5 had antibodies to thyroid gland, and 2 had malignant diseases. Of patients with foci, 26 had Helicobacter pylori–associated gastritis, 9 chronic tonsillitis or sinusitis, 4 infections with Epstein–Barr virus or cytomegalovirus, 2 dental focal infections and 2 suffered from Yersinia infection. High prevalence of H. pylori gastritis was found since 47% of patients showed elevated H. pylori–specific IgA and/or IgG antibodies. 27 patients underwent endoscopy and in all but 1 (96%) antral H. pylori infection was found. In contrast, a prevalence rate of 37% among asymptomatic adults has been published. Disappearance (67%) or improvement of urticaria (24%) occurred in most antimicrobially treated patients after 3–12 weeks. In contrast, only 50% of untreated H. pylori–seropositive patients with chronic urticaria showed spontaneous remission or improvement within 12 weeks. Prevalence of H. pylori infection may even be underestimated since only 27/100 patients underwent endoscopy. It is suggested that H. pylori infection may be present at least in all seropositive subjects (47%). Moreover, we found H. pylori infection in 2 seronegative subjects demonstrating gastric complaints. Conclusions: Thus, measurement of H. pylori–specific antibodies and/or gastroscopy should be included in the diagnostic management of chronic urticaria to identify patients who may profit from eradication treatment with disappearance of long–standing and annoying urticaria symptomatology.
Expression of B7–1, B7–2, and Interleukin 12 in Bleomycin–Induced Pneumopathy in Mice
Authors: Kaneko Yumi , Kuwano Kazuyoshi , Hagimoto Naoki , Miyazaki Hiroyuki , Kawasaki Masayuki , Fujita Masaki , Kunitake Ritsuko , Maeyama Takashige , Hara Nobuyuki
Published: April 2003

Background: The bleomycin–induced pneumopathy involves a T cell–mediated immune response. T cell activation requires both antigen/MHC recognition and costimulatory signals. The CD28 receptor on T cells with its ligand B7 represents one of the most important examples of this costimulation. Interleukin 12 (IL–12) has a strong synergistic effect with the B7–1/CD28 interaction on inducing proliferation and cytokine production in T cells. Methods: In this study, we investigated the expression of B7–1, B7–2, and IL–12 in bleomycin–induced pneumopathy in mice using reverse transcription polymerase chain reaction (RT–PCR), RT in situ PCR, and immunohistochemistry. Results: We observed concurrent upregulation of B7–1, B7–2, and IL–12p40 mRNA in the lung tissues at 1 h to 7 days after bleomycin instillation into the trachea. B7–1 mRNA and protein were found in bronchiolar epithelial cells as well as macrophages, B7–2 and IL–12p40 mRNA appeared to be expressed in mononuclear cells. Conclusions: These findings indicate that T cell–mediated immune response in this model involves the upregulation of B7–1, B7–2, and IL–12p40 mRNA, and also demonstrate the aberrant expression of B7–1 in bronchiolar epithelial cells.
Simple Understanding and Optimistic Strategy for Coping with Atopic Diseases
Authors: Okudaira Hirokazu , Mori Akio
Published: April 2003

In this article, accumulated investigations were reviewed to present an 'IL–5 central hypothesis' for eosinophilic inflammation. In this simple working hypothesis, mechanisms of clinical amelioration of atopic diseases manifested by glucocorticoids or immunosuppressants such as cyclosporin A or FK506 were easily understood to stem from the inhibition of IL–5 production and gene transcription. Screening of macrolide molecules resulted in the detection of nonactin (code name: OM–01), which inhibits production and gene transcription of IL–5 but not of IL–2 or IL–4. It was suggested that this kind of approach may open a new era of 'specific gene transcription control' for the management of allergic and other human disorders.
RANTES Production by Cytokine–Stimulated Nasal Fibroblasts: Its Inhibition by Glucocorticoids
Authors: Meyer Jens E. , Berner Ina , Teran Luis M. , Bartels Joachim , Sticherling Michael , Schröder Jens-M. , Maune Steffen
Published: April 2003

Nasal fibroblasts play an important role in both nasal polyposis and nasal allergic diseases and they are known to release a number of proinflammatory cytokines, including GM–CSF, IL–8 and IL–6. The aim of this present work was to investigate whether cytokine–stimulated nasal fibroblasts release biologically active RANTES as well as to study the effect of corticosteroids on the ability of nasal fibroblasts to produce the cytokine. Measurements of RANTES by ELISA demonstrated that RANTES is constitutively secreted spontaneously (21±4 vs. 19±6 ng/ml, respectively p>0.05). Stimulation of these cells with either TNF–α, IL–1β or IFN–γ induce further release of RANTES in a dose–dependent manner with TNF–α being the most potent stimulus. RANTES mRNA expression in nasal fibroblasts correlated with the amount of protein released in the culture supernatant upon cytokine stimulation. Moreover, chemotaxis studies demonstrated that the nasal–derived RANTES was biologically active on eosinophils. Betamethasone and hydrocortisone were found to downregulate RANTES mRNA expression in TNF–α–stimulated fibroblasts. These observations suggest that RANTES released by nasal fibroblasts may regulate eosinophil recruitment in nasal disease while glucocorticoids may inhibit the influx of these cells by suppressing the production of RANTES.
Effects of Structure Modifications on IgE Binding Properties of Serum Albumins
Authors: Restani Patrizia , Fiocchi Alessandro , Beretta Barbara , Velonà Teresa , Giovannini Marcello , Galli Corrado L.
Published: April 2003

Background: Bovine serum albumin (BSA) is one of the most widely studied proteins; its structure is well–known and its antigenic characteristics have been described in studies performed in in vitro and animal models. The aim of our work was to evaluate the role of BSA conformation in its antigenicity (recognition by circulating IgEs from allergic children). Methods: This study was performed using electrophoresis associated with the immunoblotting technique, where sera from children sensitized to BSA (as shown by double–blind placebo–controlled food challenge) were used. Results and Discussion: Heat treatment and chemical denaturation (SDS treatment) are not able to decrease the BSA capability to bind circulating IgEs. Only by reducing treatment with 2–mercaptoethanol is it possible to modify but not to eliminate the antigenicity of this protein. The reactivity to other serum albumins from different animal species was also investigated and in this study we show a direct correlation between the number of IgE–mediated responses observed in immunoblotting and the percentage of sequence identity (phylogenetic similarity) of serum albumins. Conclusion: Data obtained in this research indicate that serum albumin antigenicity is only partially correlated to its native three–dimensional structure.
Autoantibodies in Estonia and Sweden, Populations with Different Responses to Allergens
Authors: Uibo Raivo , Talja Ija , Jôgi Rain , Janson Christer , Björnsson Eythor , Boman Gunnar , Mandel Maire , Björkstén Bengt
Published: April 2003

Background: There are substantial differences in immune–mediated disease prevalence between different countries and populations which cannot be explained by genetic variability. Aims: To compare the prevalence of antinuclear (ANA), antimitochondrial, antireticulin, liver–kidney microsomal, parietal cell, thyroid microsomal and double–stranded DNA (dsDNA) antibodies among adults aged 20–44 years in two towns, i.e. in Tartu, Estonia (n = 448) and in Uppsala, Sweden (n = 532) with a low and a high prevalence of allergy. Methods: For autoantibody detection, standard indirect immunofluorescence tests on tissue and Chritidia lucilia antigenic preparations were used (sera were diluted starting from 1:10). Results: The prevalence of ANA was lower in Tartu than in Uppsala (3 vs. 10% among males and 11 vs. 16% among females; p<0.01 and p<0.1, respectively). Furthermore, dsDNA antibody–positive cases were only identified in Uppsala. The largest difference between the two populations was revealed for IgM type ANA, both among males (1 vs. 6%; p<0.02) and females (5 vs. 11%; p<0.01). In addition, the antibody titers were lower in Tartu than in Uppsala (p<0.05). The prevalence of other autoantibodies was similar in the two towns. Conclusions: The findings could support that Th2–like immune responses are commoner in Sweden than in Estonia, as previously suggested by the higher prevalence of allergy.
Flow–Cytometric Evaluation of Oxidative Burst in Phagocytic Cells of Children with Cystic Fibrosis
Authors: Frühwirth Martin , Ruedl Christiane , Ellemunter Helmut , Böck Günther , Wolf Hugo
Published: April 2003

Objectives: The objective of this study was to assess the dye 2′,7′–dichlorofluorescein (DCF) assay in screening for alterations in polymorphonuclear cell (PMN) and monocyte (MC) oxidative burst of cystic fibrosis (CF) patients. Study design: 56 CF patients aged between 2 and 20 years were investigated. Purified cells were stimulated with phorbolmyristate acetate (PMA) and zymosan (ZX). A range for DCF fluorescence for PMA– and ZX–stimulated and non–stimulated cells was established based on data from 60 healthy controls. Results: PMNs showed both enhancement and impairment. A deficient oxidative burst was detected in a total of 14 CF patients caused by abnormally high mean fluorescence intensity (MFI) of resting cells. Enhanced oxidative burst was seen in 6 CF patients. CF patients responded differently to PMA or ZX stimulation. Pseudomonas aeruginosa colonization significantly enhanced (p<0.005) the MFI of resting PMNs. MCs of CF patients showed a significantly (p<0.05) enhanced oxidative burst after stimulation with PMA compared to healthy controls, but no differences could be observed after stimulation with ZX. Serum concentrations of interleukin–6 were elevated in all CF patients, in particular in those with activation of both PMNs and MCs. Conclusion: The DCF assay shows for the first time the heterogeneity of the oxidative burst reaction in CF patients. In our opinion, the DCF assay is a reliable method for detecting pathological oxidative burst in CF patients.
Immune Response and Airway Reactivity in Wild and IL–4 Knockout Mice Exposed to Latex Allergens
Authors: Xia Jin-Qi , Rickaby David A. , Kelly Kevin J. , Choi Hongyung , Dawson Christopher A. , Kurup Viswanath P.
Published: April 2003

Background: Natural rubber latex has been reported as a major cause of allergy and asthma in a number of individuals. One of the occupational groups most affected by latex allergy are the health care workers who are frequently exposed to natural rubber latex products in their patient care activities. The immunopathogenesis of latex allergy is not well understood. In order to understand the immune mechanism in latex allergy, we have developed a mouse model of latex allergy. Methods: Both wild–type and IL–4 knockout BALB/c mice were challenged intranasally with latex proteins and their immune responses, lung pathology, and airway reactivity were evaluated. Results: The total serum IgE and latex specific IgE, IgG1, and peripheral blood and lung eosinophil levels in wild type BALB/c mice were enhanced by the latex exposure, while no IgE or eosinophil were detected in IL–4 knockout mice. Latex–specific IgG1 levels in the sera were lower in IL–4 knockout animals compared to wild mice. However, latex–specific IgG2a antibody was higher in all the IL–4 knockout mice compared to wild type mice. Both the wild type and IL–4 knockout animals developed increased airway resistance after antigen challenge when compared to control animals, although the airway resistance response of IL–4 knockout animals was attenuated compared to the wild–type animals. The histology of the lungs of these two groups of animals was similar. Conclusion: In spite of the differences in the immune responses in the two groups of mice, there were comparable lung inflammatory responses, suggesting a multifactorial pathogenetic mechanism.
Lack of Systemic Anaphylaxis and Aeroallergen–Induced Airway Plasma Extravasation in Allergic Immunoglobulin–Deficient Mice
Authors: Korsgren Magnus , Erjefält Jonas S. , Korsgren Olle , Sonmark Birgitta , Sundler Frank , Persson Carl G.A.
Published: April 2003

Background: In Ig–deficient mice allergen challenge–induced pulmonary late phase inflammation is at least as pronounced as in wild–type animals. This study investigates immediate hypersensitivity responses in these mice. Methods: To examine the acute plasma extravasation response in airway tissue, immunized Ig–deficient and wild–type mice and sham–immunized wild–type controls were subjected to 15 min ovalbumin aerosol challenge. 125I–albumin was injected (i.v.) 1 min prior to challenge. Immediately after challenge 131I–albumin was injected and the experiment was terminated. Plasma and trachea were analyzed for 125I and 131I, and the amount of extravasated plasma in the trachea was calculated. To study the development of systemic anaphylaxis immunized Ig–deficient and wild–type animals recieved intravenous allergen challenge followed by determination of mast cell responses and plasma histamine levels. Results: Allergen aerosol–exposed immunized wild–type mice exhibited marked plasma extravasation in the trachea (pd0.01 vs. wild–type controls), but in the corresponding Ig–deficient mice there was no increased extravasation. Immunized Ig–deficient mice receiving intravenous allergen challenge were resistant to anaphylactic shock. By contrast, the wild–type animals developed systemic anaphylaxis, accompanied by plasma extravasation, mast cell degranulation, elevated plasma histamine and rapid death. Conclusion: The present data are evidence that immunoglobulins are crucial for the development of immediate (type 1) responses. These findings together with our previous observations on late–phase pulmonary responses suggest that immediate hypersensitivity processes are unimportant for development of the late phase inflammation in the respiratory tract of mice.
Expression of Th1 and Th2 Immunoregulatory Cytokines by Human Eosinophils
Authors: Woerly Gaëtane , Roger Nadine , Loiseau Sylvie , Capron Monique
Published: April 2003

Besides cytotoxic mediators, human eosinophils can produce proinflammatory and anti–inflammatory cytokines, as well as growth factors and chemokines. The demonstration that eosinophils from patients could produce IL–5, IL–4 and IL–2 suggested their participation in the regulation of immune response. In the present work, we have examined the presence of Th1 (IFN–γ, IL–2) and Th2 (IL–4, IL–5, IL–10 and IL–13) cytokines in eosinophils from patients or donors by intracellular flow cytometry, and by immunocytochemistry. Whereas almost 100% eosinophils expressed intracellular IFN–γ, IL–2 and IL–10, the expression of IL–4, IL–5 and IL–13 is more variable and increased in patients versus donors. The differential release by eosinophils of Th2 versus Th1 cytokines is suggested both by the decrease of the intracellular content after culture, restricted to IL–4, IL–5 and IL–13, associated with an accumulation of IL–4 and IL–5 in the presence of Brefeldin. These results indicate that, through the preferential release of Th2 cytokines, human eosinophils can participate in the polarization of the immune response.
Stem Cell Factor–Dependent Survival, Proliferation and Enhanced Releasability of Purified Mature Mast Cells Isolated from Human Intestinal Tissue
Authors: Bischoff S.C. , Sellge G. , Schwengberg S. , Lorentz A. , Manns M.P.
Published: April 2003

Background: Recently we reported about a stem cell factor (SCF)–dependent culture system for human mast cells (MC), isolated from intestinal mucosa. Here we present a method to obtain highly purified human intestinal MC. Methods: MC were isolated from surgery specimens and purified by positive selection using the magnetic–activated cell sorting (MACS™) system and subsequent culture of the MC in medium supplemented with SCF. Results: In the presence of SCF, purified MC (50–85% purity after MACS) maintained in culture for up to 3 months. MC purity increased during culture and reached nearly 100%. During the first week of culture, MC numbers decreased, but after that time they started to proliferate. Cultured MC did not change their histamine content, phenotype or morphology. They were even more responsive towards IgE–dependent stimulation, which caused the release of high amounts of histamine, leukotrienes and cytokines such as TNF–α and IL–5. Conclusion: We show that mature human intestinal MC can be purified, maintained in culture, and triggered for proliferation in the presence of SCF. After culture, they are viable, release high amounts of mediators and cytokines upon stimulation, and thus are a valuable tool for further experiments on human mucosal MC.
IgE Inhibition as a Therapy for Allergic Disease
Authors: Jardieu Paula M. , Fick Jr. Robert B.
Published: April 2003

Monoclonal antibodies are potentially useful therapeutic agents in a variety of immunologically mediated diseases, since they offer the theoretical advantage of selectively targeting the mediators of the immuno–pathogenesis. It has been well established that IgE antibody synthesized by the immune system plays a pivotal role in the cascade of biochemical events leading to the allergic reaction. The aim of these studies was to eliminate IgE with a monoclonal antibody as the approach for treatment of atopic disease. To this end, a murine monoclonal antibody (MAE11) directed against IgE was identified which had all of the properties necessary to interfere with IgE responses. To avoid the problems of antigenicity associated with chronic administration of murine antibodies MAE11 was humanized. The best of several humanized variants, version 25 (rhuMAb–E25), was selected for clinical trials in allergic asthma and seasonal allergic rhinitis. In a series of phase I safety studies, rhuMAb–E25, by single or multidose administrations, was shown to be very well tolerated. Phase II studies were then designed to determine whether elimination of serum IgE, as a result of rhuMAb–E25 administration, had a significant impact on allergic symptoms. Results of these clinical trials establish the involvement of IgE in the pathophysiology of rhinitis and asthma and suggest a novel treatment for allergic disease.
Differential Recognition of Recombinant Phl p 5 Isoallergens by Phl p 5–Specific T Cells
Authors: van Neerven R.J.J. , Wissenbach M. , Ipsen H. , Bufe A. , Arnved J. , Würtzen P.A.
Published: April 2003

Background: At present, recombinant allergens are considered for the diagnosis and treatment of atopic allergies. To evaluate the theoretical impact of isoallergen variation on the selection of isoallergens for specific allergy vaccination, we characterized the T–cell response of allergic patients to the Phleum pratense major allergen, Phl p 5, and five of its recombinant isoallergens. Methods: Phl p 5–specific T cell lines (TCLs) were isolated from the peripheral blood of 3 allergic rhinitis patients, and their reactivity patterns were studied in detail. Results: The TCLs were highly crossreactive with related grasses. The crossreactivity with Poa pratensis was more extensive than with Lolium perenne, directly reflecting the sequence identity between Phl p 5, Poa p 5, and Lol p 5. The TCLs produced IFN–γ and IL–4 simultaneously, resembling a Th0–like cytokine production profile. Interestingly, when T cell clones were tested with natural Phl p 5 and five rPhl p 5 isoallergens, a differential recognition pattern was found. One of the TCLs exclusively reacted with Phl p 5b, another reacted with all isoforms tested, and the third reacted strongly with native purified Phl p 5, but only weakly with all five recombinant isoallergens. Conclusion: These results indicate that Phl p 5–specific T cells are highly heterogeneous, and that they differentially recognize isoallergens. This indicates that when recombinant Phl p 5 is considered for allergy vaccination, a mixture of isoallergens representing the different isoallergen groups may clinically prove to be more effective than single isoallergens.
Modulation of an Allergic Immune Response via the Mucosal Route in a Murine Model of Inhalative Type–I Allergy
Authors: Wiedermann U. , Jahn-Schmid B. , Repa A. , Kraft D. , Ebner C.
Published: April 2003

A murine model of aerosol inhalation, leading to sensitization to birch pollen (BP) and its major allergen Bet v 1, was established in order to try to influence type–I allergic immune responses via the mucosal route. We previously demonstrated that simultaneous inhalation of BP and cholera toxin, a potent mucosal adjuvant, induced a Th1–like immune response to the allergen in naive mice and modulated allergic immune responses in sensitized mice. In contrast to cholera holotoxin, mucosal application of the cholera B subunit (CTB) conjugated to antigen has been shown to induce peripheral tolerance in certain models of Th1–based autoimmune diseases. In the present study we investigated the potential of such an antigen delivery system to suppress Th2–based, allergic immune responses. Mucosal administration of CTB/Bet v 1 conjugates prior to sensitization led to significantly increased allergen–specific IgE/IgG1 and IgG2a antibody levels and cytokine production (IL–5, IFN–γ) in vitro. Thus, CTB coupled to Bet v 1 acted as an adjuvant rather than a tolerogen. On the other hand we noted that mucosal application of CTB coupled to ovalbumin led to marked suppression of antigen–specific IgE antibody levels and IL–5 production in vitro and thereby restricted allergic sensitization. These results indicated that the effects of CTB/antigen conjugates depended on the nature of the antigen. In contrast to Bet v 1 coupled to CTB, nasal as well as oral application of low doses of unconjugated, Bet v 1 prior to aerosol sensitization inhibited allergen–specific antibody responses of all isotypes, cutaneous type–I skin tests in vivo as well as allergen–specific lymphoproliferative responses and cytokine production (IL–4, IL–5, IL–10, IFN–γ) in vitro, suggesting that both T– and B–cell tolerance to the allergen were induced. Taken together, mucosal tolerance induction as well as the use of certain transmucosal antigen delivery systems might be promising new strategies to modulate type–I allergic immune responses
Nerve Growth Factor: An Important Molecule in Allergic Inflammation and Tissue Remodelling
Authors: Bonini S. , Lambiase A. , Bonini S. , Levi-Schaffer F. , Aloe L.
Published: April 2003

Background: Several studies provide evidence that nerve growth factor (NGF) has an expanding role in neuroimmune interactions. Methods: We review our data on circulation levels of NGF in allergic diseases as well as on the relationships between this neurotrophin and primary and secondary effector cells of allergic inflammation. Results: In vernal keratoconjunctivitis, a close relationship exists between the increased NGF plasma values and the number of mast cells infiltrating the conjunctiva. NGF serum values are also increased in other allergic diseases and asthma, and are related to the severity of the inflammatory process and disease. Human CD4+ T cell clones (preferentially of activated Th2 type) produce and release NGF, and express high–affinity NGF receptors. NGF is preformed in and can act on human peripheral blood eosinophils to preferentially release inflammatory mediators. Immunoreactivity for high affinity NGF receptors is present both in basal epithelial cells and in the inflamed stroma of the allergic conjunctiva. Topical administration of NGF results in a complete healing of neurotophic corneal ulcers in man, thus suggesting a profound effect of NGF on human fibroblasts and extracellular matrix. Conclusion: Data presented suggest that NGF is an important molecule in allergic inflammation and tissue remodelling occurring in allergic diseases.
Neurotrophins: A Link between Airway Inflammation and Airway Smooth Muscle Contractility in Asthma?
Authors: Braun Armin , Lommatzsch Marek , Lewin Gary R. , Virchow Johann C. , Renz Harald
Published: April 2003

Background: Bronchial asthma (BA) is characterized by a unique type of airway inflammation, epithelial cell damage and increased airway smooth muscle (ASM) contractility. The regulatory network between the immunological events and the neuronal control of ASM contractility remains to be defined. Methods: Utilizing a well–characterized mouse model of airway inflammation and BA, we analyzed the production and function of neurotrophins in allergic asthma. To confirm these data in humans, segmental allergen provocation was performed in mild asthmatics. Results: Allergen–induced airway inflammation was associated with increased local production of the neurotrophins nerve growth factor (NGF) and brain–derived neurotrophic factor (BDNF) in mice as well as in humans. In bronchoalveolar lavage fluid (BALF), NGF levels were increased 4– to 5–fold in men and mice 1 day after allergen provocation. The increase in BDNF was about 2–fold in both models. Treatment of mice with anti–NGF prevented development of airway hyperresponsiveness (AHR). In the human study group, NGF levels in BALF after allergen provocation were correlated significantly with baseline FEV1 levels. Conclusion: These data strongly suggest that neurotrophins serve as a link between airway inflammation and neuronal control of ASM constriction in BA.
Role of the Eosinophil in Protein Oxidation in Asthma: Possible Effects on Proteinase/Antiproteinase Balance
Authors: Foreman Richard C. , Mercer Paul F. , Kroegel Claus , Warner Jane A.
Published: April 2003

Background: Many of the leukocytes which migrate into the tissue following allergen challenge can undergo a respiratory burst producing reactive oxygen species causing tissue damage and distorting proteinase/antiproteinase balance. The reactive oxygen species have extremely short half lives and so cannot be measured in vivo, but the protein carbonyl residues which result from protein oxidation can be measured in biological fluids. Methods: We examined protein oxidation in bronchoalveolar lavage after allergen challenge in 12 patients with atopic asthma by measuring protein carbonyl residues using a sensitive Western blotting technique. Results: We found that the median level of protein carbonyls per molecule of protein rose from 0.09 in bronchoalveolar lavage (BAL) obtained 18 h after saline challenge to 0.23 in BAL taken 10 min after segmental allergen challenge, reaching a median of 0.82 in samples obtained 18 h later (p<0.01 compared with saline control and the earlier time point). The number of protein carbonyl residues correlated strongly with the number of eosinophils recovered in the BAL (rho = 0.574, p<0.05) but not with the number of neutrophils (rho = 0.228, p = NS) or macrophages (rho = 0.178, p = NS). Western blotting showed that the majority of the modified protein comigrated with authentic α1–antitrypsin at 53 kD. In contrast, BAL samples from patients with chronic obstructive pulmonary disease, which is characterized by an influx of neutrophils, showed that the main oxidized protein colocalized with human serum albumin, the predominant protein in the BAL. Conclusion: Our results suggest that the recruitment and activation of eosinophils accounts for much of the protein oxidation found in the BAL following allergen challenge. The main oxidized protein appears to be α1–antitrypsin, a key antiproteinase in the airways. Inactivation of α1–antitrypsin by oxidation may distort the proteinase/antiproteinase balance leaving the lung vunerable to proteolytic damage. Intriguingly, we have evidence to suggest that other airways diseases, characterized by recruitment of other inflammatory cells, may result in the oxidation of alternative targets.
Endogenous Superallergen Protein Fv Interacts with the VH3 Region of IgE to Induce Cytokine Secretion from Human Basophils
Authors: Patella Vincenzo , Giuliano Ada , Florio Giovanni , Bouvet Jean-Pierre , Marone Gianni
Published: April 2003

Background: Protein Fv is an endogenous protein, synthesized in liver and largely released in the digestive tract during acute and chronic viral hepatitis, that binds to immunoglobulin (Ig) from various mammalian and nonmammalian species. Methods: Basophils obtained from normal subjects were purified by a double Percoll gradient and elutriation. The secretion of histamine induced by protein Fv was assayed by a fluorometric technique, the extracellular protein levels of IL–4 and IL–13 were measured by ELISA, and IL–4 mRNA levels were evaluated by RT–PCR. Results: Protein Fv concentration–dependently induced histamine and IL–4 and IL–13 release from purified basophils. IL–4 mRNA, constitutively present in basophils, was increased after stimulation by protein Fv. Histamine and IL–4 secretion from basophils, but not histamine and IL–13 release, activated by protein Fv was significantly correlated (rs = 0.70, p<0.001). Basophils from which IgE had been dissociated by brief exposure to lactic acid no longer released IL–4 in response to protein Fv and anti–IgE. Two preparations of human VH3+ monoclonal IgM inhibited protein Fv–induced secretion of IL–4 and histamine from basophils. In contrast, VH6+ monoclonal IgM did not inhibit the release of mediators caused by protein Fv. Conclusions: These results indicate that protein Fv, which acts as an endogenous superallergen, interacts with the VH3 domain of IgE to induce the synthesis and release of IL–4, IL–13 and the secretion of histamine from basophils.
Genetic Determinants of 5–Lipoxygenase Transcription
Authors: Drazen Jeffrey M. , Silverman Eric S.
Published: April 2003

Background: 5 Lipoxygenase (5–LO) is a critical enzyme in the production of the leukotrienes. We have identified a series of mutations in the 5–LO gene that modify gene transcription. These mutations consist of addition of an Sp–1 binding motif (–GGGCGG–) or deletion of one or two Sp–1 binding motifs in the 5–LO core promoter. Methods: Mutant forms of the 5–LO core promoter were placed in a chloramphenicol acetyl transferase (CAT) reporter construct using either HeLa or SL–2 cells. Results: In HeLa cells all of the mutant forms are less effective in driving CAT reporter activity than the wild–type promoter. In SL–2 cells the construct containing the addition mutation was more effective in driving CAT reporter activity, while the constructs containing the deletion mutations were less effective. Conclusions: These data indicate that naturally occurring mutations in the 5–LO core promoter modify gene transcription in vitro.
Analysis of γ4 Germline Transcription in Human B Cells
Authors: Agresti Alessandra , Vercelli Donata
Published: April 2003

Background: Allergic reactions occur rarely in patients with chronic helminth infections, although FcεRI–bearing cells are sensitized with anti–parasite IgE and are exposed to antigen. The inhibition of allergic reactivity is due to ‘blocking antibodies’, predominantly IgG4. IgG4 antibodies represent 50–95% of anti–filarial IgG, and have blocking activity because they compete with cell–bound IgE for allergen binding. Blocking IgG4 antibodies have also been detected in patients treated with immunotherapy. However, the molecular mechanisms of IgG4 regulation have remained unexplored. We address this issue starting with the IL–4–dependent induction of γ4 germline transcription, an essential step in the commitment to isotype switching. Results: γ4 germline transcripts were cloned and shown to contain Iγ4 spliced to the 5′ portion of Cγ4 using the splice donor site shared by γ1 and γ3 germline transcripts. Sequence analysis located the human Iγ4 exon ≈0.6 kb upstream of the Sγ4 region. Four major transcription start sites located 547–583 bp upstream of the Iγ4 splice donor site were identified by primer extension analysis. A HindIII/NaeI construct (–413/+466) had the highest activity in reporter assays. Transcription was induced 4.6–fold by IL–4, 2.1–fold by CD40 engagement, and 14.5–fold by a combination of the two signals. Thus CD40 crosslinking enhanced IL–4–dependent transcription by 3.1–fold, showing that the two stimuli act synergistically. Conclusion: As we understand more about IgG4 regulation, our hope is to apply to allergy the lesson we learnt from helminth infections.
Timothy Grass (Phleum pratense L.) Pollen as Allergen Carriers and Initiators of an Allergic Response
Authors: Behrendt H. , Tomczok J. , Sliwa-Tomczok W. , Kasche A. , Ebner von Eschenbach C. , Becker W.M. , Ring J.
Published: April 2003

Contrary to indoor allergen exposure (e.g. house dust mite), there is no reliable quantitative association between pollen exposure and symptoms of allergic diseases. Therefore we studied localization and release of major allergens from timothy grass (Phleum pratense L.) pollen using different methods and pollen grain sources. Localization of major allergens Phl p 5 and Phl p 1 was visualized by field emission scanning electron microscopy after anhydrous fixation and immunogold silver staining in a three–dimensional reconstruction; Phl p 5 was found in the cytoplasm and on the exine, Phl p 1 in the intine. No allergens were found inside the starch granules. Allergen liberation from pollen grains was studied in vitro under physiological conditions (30 min, 37°C) at pH 6.0, 7.4 and 9.0. Besides total protein measurements in the supernatant, major allergens were determined by immunoblot, Phl p 5 was quantitated by ELISA. There were striking differences in total protein and major allergen release between freshly collected and commercially available grass pollen grains as well as among freshly collected pollen between rural meadows and areas near high–traffic roads. There was a significantly different release of total protein being lowest in supernatants from commercially available pollen grains (rural/traffic vs. commercial, p<0.001), and of Phl p 5 major allergen (rural>traffic>commercial, p<0.005). Therefore, allergen bioavailability seems to be an important parameter in order to establish reliable dose–response relationships for the outdoor allergen response. Pollen grains incubated in aqueous protein–free buffer solution were also found to secrete significant amounts of eicosanoids namely prostaglandin E2 and leukotriene B4. Pollen grains thus do not act only as allergen carriers but also might have important implications on early events as initiators of allergy.
Expression of the T–Cell Markers CD3, CD4 and CD8 in Healthy and Atopic Children during the First 18 Months of Life
Authors: Aniansson Zdolsek Helena , Ernerudh Jan , Holt Patrick G. , Nilsson Joacim , Björkstén Bengt
Published: April 2003

Background: There is little information available about the development of T–cell immunity in healthy and atopic children. We have studied prospectively the mean fluorescence intensity of the T–cell receptor complex–associated CD3, CD4 and CD8 in relation to atopic family history (AFH) and the development of atopic disease. Methods: Children with a defined AFH (n = 172) were followed from birth to 18 months and the cumulative history of atopic disease was recorded. Blood samples were obtained at birth and at 18 months, and in a subgroup of 78 children also at 3, 6 and 12 months. Multicolour flow cytometry was used to analyse pan T–cells (CD3+CD45+CD14–), T–helper–(CD3+CD4+) and T–cytotoxic–(CD3+CD8+) cells. Results: At 18 months, 31 children were atopic and 118 non–atopic. Children who developed atopic disease had a higher CD4 expression (mean fluorescence intensity, MFI) on CD4+CD3+ lymphocytes at birth and at 3 months, particularly as compared with non–atopic children without AFH. Furthermore, the CD3 expression on CD3+CD45+CD14– lymphocytes increased more slowly with age in children with double atopic heredity, as compared with children with no or only one atopic family member. Conclusions: The higher expression of the CD4 receptor in early infancy in children who developed atopic disease compared with non–atopics suggests a delayed expression in T–helper cells. Children with a strong AFH had a slower increase in the expression of CD3, indicating a delayed T–cell maturation.
Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk
Authors: Papadopoulos Nikolaos G. , Syrigou Ekaterini I. , Bossios Apostolos , Manou Orsalia , Gourgiotis Dimitrios , Saxoni-Papageorgiou Photini
Published: April 2003

Background: Controversial results have been reported on the participation and diagnostic value of lymphocyte reactivity in cow’s milk (CM) allergy. In this study, we used a specific nuclear marker to evaluate lymphocyte proliferation in IgE–mediated CM allergy in infants, and examine its relation with diets containing different CM antigen loads. Methods: Infants with IgE–mediated CM allergy, as assessed by open provocation and RAST, were grouped according to their exclusive diet, either CM formulae, breast feeding, or hydrolysed whey formulae. A group of non–atopic infants receiving CM was also examined. Lymphocyte proliferation to β–lactoglobulin was evaluated by quantitation of the proliferating cell nuclear antigen (PCNA) expression in peripheral blood mononuclear cells, by flow cytometry. Immunophenotypic surface markers were also examined. Results: A marked difference of PCNA expression between CM–fed allergic infants and healthy controls was observed (p<0.001). In this setting, PCNA expression ≥10% was highly specific and sensitive as a marker of CM allergy in CM–fed infants. Moreover, a significant correlation (p<0.001) between antigen load and PCNA was established in CM–allergic infants under different diets, higher values obtained with increasing antigen loads. In addition, within the group fed hydrolyzed formulae, low–molecular–weight products resulted in marginally lower PCNA expression than higher–molecular–weight formulae. No differences in immunophenotype were found, with the exception of a higher CD23 expression in the breast–fed group. Conclusions: PCNA could be a useful marker in the assessment of lymphocyte proliferation to CM antigens. Low CM antigen diets are related with reduced lymphocyte reactivity, which may partly explain the clinical benefit observed with such diets.
Early Interleukin–4: Its Role in the Switch towards a Th2 Response and IgE–Mediated Allergy
Authors: Haas H. , Falcone F.H. , Holland M.J. , Schramm G. , Haisch K. , Gibbs B.F. , Bufe A. , Schlaak M.
Published: April 2003

The etiology of IgE–mediated allergies is complex and, thus far, not completely understood. A common feature, however, is the overproduction of IgE–inducing cytokines, e.g. interleukin–4(IL–4), compared to IgE–antagonistic cytokines, such as interferon–γ or IL–12. IgE–inducing cytokines are produced by T helper type 2 (Th2) cells. The differentiation of naive T cells towards the Th2 phenotype seems to be crucially dependent upon the particular cytokines present in the early stages of an immune response. Concerning the factors driving Th2 differentiation, the so–called 'early IL–4’ seems to play an important role, although there is some controversy over the degree of its requirement and its cellular source. We have recently demonstrated that basophils might be such a source, since they rapidly release IL–4 upon antigen–specific or nonantigen–specific stimuli, such as certain lectins. This makes lectins interesting candidates for inducing a Th2 response and IgE–mediated allergy in unsensitized individuals.
molecular Cloning of the Guinea Pig GRO Gene and Its Rapid Expression in the Tissues of Lipopolysaccharide–Injected Guinea Pigs
Authors: Yoshimura Teizo , Takeya Motohiro , Ogata Hiroomi , Yamashiro Shigeo , Modi William S. , Gillitzer Reinhard
Published: April 2003

Background: CXC chemokines, IL–8 and GRO, play a role in the recruitment of neutrophils in the human. The functional orthologues in the rat and mouse are CINC/KC and MIP–2. The lack of IL–8 made these animals less useful to study the role of IL–8 and GRO. Methods:  Guinea pig (gp) cDNA libraries were screened for GRO and IL–1β. A gp genomic library was screened with a gpGRO cDNA probe. Expression of gpIL–8, gpGRO, gpTNFα, and gpIL–1β was investigated by Northern analysis and/or by in situ hybridization. Results: Two gpGRO cDNAs, a 3.0–kb gpGRO genomic DNA, and a gpIL–1β cDNA were cloned. gpGRO and gpIL–8 mRNA were detected in different tissues including lungs 1 h after intraperitoneal injection of lipopolysaccharide (LPS) into guinea pigs. gpGRO, gpIL–8, gpTNFα, and gpIL–1β expression peaked at 3 h in the lungs. Both gpGRO and gpIL–8 mRNA were detected in the cells in alveolar spaces and bronchial epithelial cells. However, gpGRO mRNA, but not gpIL–8, was also expressed in endothelial cells and vascular smooth muscle cells. Conclusions: gpGRO and gpIL–8 mRNA rapidly accumulated in the lungs of guinea pigs after LPS injection. Expression of gpIL–8 and gpGRO mRNA appeared to be independent from TNFα– or IL–1β–stimulation in this model. A high level expression of gpGRO in vascular cells suggest an important role of GRO in the sequestration of neutrophils and multi–organ injuries induced by LPS. The guinea pig will provide an excellent model to study the roles of IL–8 and GRO, important inflammatory mediators in the human.
Functional Cutaneous Lymphocyte Antigen Can Be Induced in Essentially All Peripheral Blood T Lymphocytes
Authors: Armerding Dieter , Kupper Thomas S.
Published: April 2003

The cutaneous lymphocyte–associated antigen (CLA) is a skin–homing receptor expressed on a minority of memory–type peripheral blood T (PBT) lymphocytes. Induction of high–level CLA expression in PBT has previously been difficult to accomplish in vitro. Here we report that constitutive CLA expression could be readily induced in virtually all PBT by various polyclonal activators. There was no requirement for accessory cells or addition of other mediators except for IL–2 for maintaining cell survival. Absence of serum in the culture medium was important for optimal induction of CLA. The number of T cells adhering to E–selectin as well as tethering and shear stress resistance under hydrodynamic flow increased in correlation with the level of cell surface CLA expressed. Clonal analysis of CLA induction revealed that in serum–containing medium, which permits the majority of T cells to expand, only a minority of clones did not express CLA. Such T cells could be induced to highly express CLA within 8 days by switching from serum–containing to serum–free medium. This cell–surface phenotype change was closely associated with acquisition of E–selectin ligand activity. Fucosyltransferase VII, which is believed to be important for the generation of the CLA epitope on the P–selectin glycoprotein ligand–1 (PSGL–1) backbone, was shown to be significantly increased in CLA–positive versus CLA–negative T cell populations by PCR analysis. Our findings are consistent with the idea that restriction of CLA expression after activation, rather than positive selection of predetermined T cell subpopulations exposed to restrictive stimulatory conditions in unique microenvironments, may be important in vivo.