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Der P (der + p)

Distribution by Scientific Domains
Medical Sciences92%
Life Sciences4%

Kinds of Der P

  • allergen der p
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  • mite allergen der p
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    Selected Abstracts

    Variability of IgE reactivity profiles among European mite allergic patients

    EUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 12 2008
    M. Weghofer
    Abstract Background, House dust mites (HDM) Dermatophagoides pteronyssinus are a frequent indoor allergen source. Our aim was to determine the frequencies of IgE reactivity to purified HDM allergen molecules in mite allergic patients from different parts of Europe in order to establish an allergen panel for diagnosis of HDM allergy. Materials and methods, Populations of D. pteronyssinus -allergic patients from Austria (n = 56), France (n = 55), Italy (n = 67) and Sweden (n = 65) and storage mite allergic patients from Sweden (n = 31) were analysed for IgE reactivity to eight purified natural (n) and recombinant (r) D. pteronyssinus allergens (nDer p 1, rDer p 2, nDer p 4, rDer p 5, rDer p 7, rDer p 8, rDer p 10 and rDer p 14) in RAST-based dot blot assays. Results, Using a combination of Der p 1 and Der p 2, at least 97% of the D. pteronyssinus -allergic patients could be diagnosed in each of the HDM allergic populations. However, more than 50% of the patients also reacted with other allergens and significant variabilities regarding the frequencies of IgE reactivity to individual allergen molecules were found. Patients with a predominant storage mite allergy showed none or only very weak IgE reactivity to purified D. pteronyssinus allergens. Conclusions, Purified Der p 1 and Der p 2 are sufficient for the diagnosis of , 97% of D. pteronyssinus allergic patients in Europe, but other allergens may also play an important role for the diagnosis and treatment of HDM allergy. [source]

    Lipopolysaccharide binding of the mite allergen Der f 2

    GENES TO CELLS, Issue 9 2009
    Saori Ichikawa
    Lipid-binding properties and/or involvement with host defense are often found in allergen proteins, implying that these intrinsic biological functions likely contribute to the allergenicity of allergens. The group 2 major mite allergens, Der f 2 and Der p 2, show structural homology with MD-2, the lipopolysaccharide (LPS)-binding component of the Toll-like receptor (TLR) 4 signalling complex. Elucidation of the ligand-binding properties of group 2 mite allergens and identification of interaction sites by structural studies are important to explore the relationship between allergenicity and biological function. Here, we report a ligand-fishing approach in which His-tagged Der f 2 was incubated with sonicated stable isotope-labelled Escherichia coli as a potential ligand source, followed by isolation of Der f 2-bound material by a HisTrap column and NMR analysis. We found that Der f 2 binds to LPS with a nanomolar affinity and, using fluorescence and gel filtration assays that LPS binds to Der f 2 in a molar ratio of 1 : 1. We mapped the LPS-binding interface of Der f 2 by NMR perturbation studies, which suggested that LPS binds Der f 2 between the two large ,-sheets, similar to its binding to MD-2, the LPS-binding component of the innate immunity receptor TLR4. [source]

    Single nucleotide polymorphisms and haplotype of MD-1 gene associated with high serum IgE phenotype with mite-sensitive allergy in Taiwanese children

    INTERNATIONAL JOURNAL OF IMMUNOGENETICS, Issue 6 2007
    J. Y. Wang
    Summary MD-1 (myeloid differentiation 1; also known as Ly86, lymphocyte antigen 86), interacting with RP105, plays an important role in Toll-like receptor 4 (TLR4) signalling pathway. It has been suggested to be involved in the pathological mechanism of inflammation and atopic diseases. The purpose of this study was to investigate the genetic association between single nucleotide polymorphisms (SNPs) of MD-1 promoter and coding region and mite-sensitive allergy in Taiwanese children. We conducted a case-control study on 237 controls and 281 allergic patients sensitive to Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) by genotyping 35 SNPs in MD-1 gene region. In the promoter region we identified three SNPs, rs1334710, rs4959389, and rs977785 that are associated with mite-sensitive allergy in Taiwanese children. The P -values ranged from 0.0150 to 0.009. The haplotypes including promoter region were also associated with mite-sensitive allergy. Our results suggested that MD-1 could be a susceptible gene for mite-sensitive allergy in Taiwanese children. [source]

    Changes in Cell Wall-bound Phenolic Compounds and Lignin in Roots of Date Palm Cultivars Differing in Susceptibility to Fusarium oxysporum f. sp. albedinis

    JOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2000
    C. El Modafar
    The roots of date palm contain four cell wall-bound phenolic acids identified as p -hydroxybenzoic, p -coumaric, ferulic and sinapic acids. The ferulic acid represents the major phenolic compound since it constitutes 48.2,55.8% of cell wall-bound phenolic acids. All these phenolic acids were present in the resistant cultivar (BSTN) and the susceptible cultivar (JHL). However, the pre-infection contents of p -coumaric, ferulic and sinapic acids were greater in the resistant cultivar than in the susceptible one. For the contents of p -hydroxybenzoic acid, there was no significant difference between the resistant cultivar and the susceptible cultivar. Similarly, the pre-infection contents of lignin were approximately equal for both cultivars. Inoculation of the date palm roots by Fusarium oxysporum f. sp. albedinis induced important modifications to the contents of the cell wall-bound phenolic compounds and lignin, which made it possible to distinguish between resistant and susceptible cultivars. The post-infection contents of cell wall-bound phenolic compounds underwent a rapid and intense increase with a maximum accumulation on the tenth day for p -hydroxybenzoic acid (1.54 ,mol/g), p -coumaric acid (2.77 ,mol/g) and ferulic acid (2.64 ,mol/g) and on the fifteenth day for sinapic acid (1.85 ,mol/g). The maximum contents accumulated in the resistant cultivar were greater than those in the susceptible cultivar, namely, 11 times for p -hydroxybenzoic acid, 2.6 times for p -coumaric acid, 1.8 times for ferulic acid and 12.3 times for sinapic acid. In the susceptible cultivar, p -coumaric acid and ferulic acid contents also increased after inoculation although they did not reach the pre-infection contents of the resistant cultivar. The contents of p -hydroxybenzoic acid in the susceptible cultivar roots did not present post-infection modification and those of sinapic acid decreased instead. The lignin contents increased in both cultivars with a maximum accumulation on the fifteenth day. However, the maximum contents accumulated in the resistant cultivar roots were 1.5 times greater than those of the susceptible cultivar. These results showed clear differences between the resistant BSTN and the susceptible JHL cultivars. The implication of cell wall-bound phenolic compounds and lignin in the resistance of date palm to F. oxysporum f. sp. albedinis appears to be dependent on the speed and intensity of their accumulation with greater contents in the first stage of infection. Zusammenfassung Die Wurzeln der Dattelpalme enthalten vier zellwandgebundene Phenolsäuren, die als p -Hydroxybenzoesäure, p- Cumarsäure, Ferulasäure und Sinapinsäure identifiziert wurden. Ferulasäure ist die wichtigste phenolische Verbindung, denn sie stellt 48,2 bi 55,8% der zellwandgebundenen Phenolsäuren. Alle vier Phenolsäuren waren in der resistenten Sorte BSTN und in der anfälligen Sorte JHL vorhanden. Die Gehalte an p -Cumarsäure, Ferulasäure und Sinapinsäure waren vor der Infektion bei der resistenten Sorte jedoch höher als bei der anfälligen Sorte. Hinsichtlich des Gehalts an p -Hydroxybenzoesäure unterschieden sich die resistente und die anfällige Sorte nicht signifikant voneinander. Auch die Ligningehalte vor der Infektion waren bei beiden Sorten ungefähr gleich. Die Inokulation von Dattelpalmenwurzeln mit Fusarium oxysporum f. sp. albedinis induziert wichtige Änderungen der Gehalte an zellwandgebundenen phenolischen Verbindungen und Lignin, was eine Unterscheidung resistenter von anfälligen Sorten ermöglicht. Nach der Infektion nehmen die Gehalte an zellwandgebundenen phenolischen Verbindungen rasch und erheblich zu, mit maximaler Akkumulation am 10. Tag bei p -Hydroxybenzoesäure (1,54 ,mol/g), p -Cumarsäure (2,77 ,mol/g) und Ferulasäure (2,64 ,mol/g) sowie am 15. Tag bei Sinapinsäure (1,85 ,mol/g). Die in der resistenten Sorte akkumulierten maximalen Gehalte waren höher als die maximalen Gehalte der anfälligen Sorte , um das Elffache bei p -Hydroxybenzoesäure, das 2,6-fache bei p -Cumarsäure, das 1,8-fache bei Ferulasäure und das 2,3-fache bei Sinapinsäure. Bei der anfälligen Sorte steigen die Gehalte an p -Cumarsäure und Ferulasäure nach der Inokulation ebenfalls an, bleiben aber niedriger als die Werte der resistenten Sorte vor der Infektion. Der p -Hydroxybenzoesäuregehalt in den Wurzeln der anfälligen Sorte ist nach der Infektion nicht verändert, und der Sinapinsäuregehalt nimmt ab. Der Ligningehalt steigt bei beiden Sorten, mit maximaler Akkumulation am 15. Tag. Der maximale Gehalt in den Wurzeln der resistenten Sorte war jedoch 1,5-mal höher als bei der anfälligen Sorte. Diese Ergebnisse zeigen deutliche Unterschiede zwischen der resistenten Sorte (BSTN) und der anfälligen Sorte (JHL). Die Bedeutung der zellwandgebundenen phenolischen Verbindungen und des Lignins für die Resistenz der Dattelpalme gegen F. oxysporum f. sp. albedinis scheint von der Geschwindigkeit und der Intensität der Akkumulation abzuhängen, wobei die Gehalte im ersten Stadium der Infektion höher sind. [source]

    Major mite allergen Der f 1 concentration is reduced in buildings with improved energy performance

    ALLERGY, Issue 5 2010
    F. Spertini
    To cite this article: Spertini F, Berney M, Foradini F, Roulet C-A. Major mite allergen Der f 1 concentration is reduced in buildings with improved energy performance. Allergy 2010; 65: 623,629. Abstract Background:, Environmental conditions play a crucial role in mite growth, and optimal environmental control is key in the prevention of airway inflammation in chronic allergic rhinoconjunctivitis or asthma. Objective:, To evaluate the relationship between building energy performance and indoor mite allergen concentration in a cross-sectional study. Methods:, Major allergen concentration (Der f 1, Der p 1, mite group 2, Fel d 1 and Bla g 2) was determined by quantitative dot blot analysis from mattress and carpet dust samples in five buildings designed for low energy use (LEB) and in six control buildings (CB). Inhabitants had received 4 weeks prior to mite measurement a personal validated questionnaire related to the perceived state of health and comfort of living. Results:, Cumulative mite allergen concentration (with Der f 1 as the major contributor) was significantly lower in LEB as compared with CB both in mattresses and in carpets. In contrast, the two categories of buildings did not differ in Bla g 2 and Fel d 1 concentration, in the amount of dust and airborne mould collected. Whereas temperature was higher in LEB, relative humidity was significantly lower than in CB. Perceived overall comfort was better in LEB. Conclusions:, Major mite allergen Der f 1 preferentially accumulates in buildings not specifically designed for low energy use, reaching levels at risk for sensitization. We hypothesize that controlled mechanical ventilation present in all audited LEB may favour lower air humidity and hence lower mite growth and allergen concentration, while preserving optimal perceived comfort. [source]

    Characterization and comparison of commercially available mite extracts for in vivo diagnosis

    ALLERGY, Issue 2 2010
    B. Brunetto
    To cite this article: Brunetto B, Tinghino R, Braschi MC, Antonicelli L, Pini C, Iacovacci P. Characterization and comparison of commercially available mite extracts for in vivo diagnosis. Allergy 2010; 65: 184,190. Abstract Background:, Assessment of sensitization by allergen-specific IgE testing and skin prick testing (SPT) are primary tools in routine clinical diagnosis of allergies. To perform a correct diagnosis, it is critical that the allergen reagent used contains an adequate amount of all relevant components. This study aimed at evaluating commercially available mite extracts for in vivo diagnosis from eight manufacturers. Methods:, Eight extracts from Dermatophagoides pteronyssinus and eight from Dermatophagoides farinae were analysed for total protein content by Bradford and for major allergen content by ELISA. SDS-PAGE, immunoblotting and SPT were also carried out. Results:, The protein amount ranged from 27.7 ,g/ml extract to 361.1 ,g/ml (D. pteronyssinus) and from 20.3 to 353.0 ,g/ml (D. farinae). In regards major allergen concentration, Der p 1 ranged from 9.6 to 36.2 ,g/ml, Der f 1 26.5,196.1 ,g/ml, mite group 2 0.7,31.7 ,g/ml in D. pteronyssinus and 1.3,10.4 ,g/ml in D. farinae. SDS-PAGE experiments showed that some components are poorly represented or absent in extracts from most manufacturers. Similar results were obtained by IgE-immunoblotting and SPT with 10 mite allergic patients confirmed a broad spectrum of reactivity of the extracts in the same subject. Conclusions:, Immunochemical analysis showed a heterogeneous amount of component/s among mite extracts from different manufacturers. These data were confirmed by in vivo testing, suggesting that, for some of the patient tested, the absence of relevant allergens could strongly affect the diagnosis. [source]

    Effect of improved home ventilation on asthma control and house dust mite allergen levels

    ALLERGY, Issue 11 2009
    G. R. Wright
    Background:, The warm, humid environment in modern homes favours the dust mite population, but the effect of improved home ventilation on asthma control has not been established. We tested the hypothesis that a domestic mechanical heat recovery ventilation system (MHRV), in addition to allergen avoidance measures, can improve asthma control by attenuating re-colonization rates. Methods:, We conducted a randomized double-blind placebo-controlled parallel group trial of the installation of MHRV activated in half the homes of 120 adults with asthma, allergic to Dermatophagoides pteronyssinus. All homes had carpets steam cleaned and new bedding and mattress covers at baseline. The primary outcome was morning peak expiratory flow (PEF) at 12 months. Results:, At 12 months, the primary end-point; change in mean morning PEF as compared with baseline, did not differ between the MHRV group and the control group (mean difference 13.5 l/min, 95% CI: ,2.6 to 29.8, P = 0.10). However, a secondary end-point; evening mean PEF, was significantly improved in the MHRV group (mean difference 24.5 l/min, 95% CI: 8.9,40.1, P = 0.002). Indoor relative humidity was reduced in MHRV homes, but there was no difference between the groups in Der p 1 levels, compared with baseline. Conclusions:, The addition of MHRV to house dust mite eradication strategies did not achieve a reduction in mite allergen levels, but did improve evening PEF. [source]

    Mite serine protease activates protease-activated receptor-2 and induces cytokine release in human keratinocytes

    ALLERGY, Issue 9 2009
    T. Kato
    Background:, House dust mites produce serine and cysteine proteases. Mite-derived proteases have been suggested to be involved in the pathogenesis of allergies; however, whether mite-derived serine protease activity can stimulate keratinocytes remains unknown. Methods:, We examined the activation of primary human keratinocytes by serine protease-rich extract of whole mite culture and compared with that by recombinant group 1 allergens (rDer f 1 and rDer p 1), which exclusively exhibit cysteine protease activity. Results:, Protease activity of whole mite culture extract (WCE), rDer f 1 and rDer p 1 induced the release of IL-8 and granulocyte-macrophage colony-stimulating factor. Protease activity of WCEs induced a significant upregulation of their mRNA expression but rDer f 1 had much less effect. Protease activity of the WCE stimulated intracellular Ca2+ mobilization but rDer f 1 and rDer p 1 did not. The mobilization induced by agonists for the human protease-activated receptor (PAR)-2, an agonist peptide or trypsin, was diminished by pre-incubation of keratinocytes with WCE. rDer f 1 inefficiently cleaved a synthetic N-terminal peptide of PAR-2 at different sites from trypsin, but the resultant peptides did not stimulate the release of interleukin-8. Conclusions:, The results suggest that mite-derived serine protease activity may contribute to the pathogenesis of atopic dermatitis by activating keratinocytes via PAR-2 activation but cysteine protease activity of Der f 1 and Der p 1 acts via another mechanism. [source]

    Immunomodulatory properties of Lactobacillus plantarum and its use as a recombinant vaccine against mite allergy

    ALLERGY, Issue 3 2009
    P. Rigaux
    Background:, Selected lactic acid bacteria were reported to prevent atopic dermatitis and experimental asthma but the mechanisms of their immunomodulatory effects are not fully elucidated. In this study, the signaling pathways triggered by Lactobacillus plantarum NCIMB8826 were investigated and the potential use of this strain producing a variant of the mite allergen Der p 1 as live vaccine vehicle was evaluated. Methods:, Mouse bone marrow-derived dendritic cells were stimulated with wild-type or a L. plantarum teichoic acid mutant to evaluate the secretion of cytokines. A recombinant L. plantarum expressing Der p 1 was engineered, its in vitro immunomodulatory properties were characterized and its prophylactic potential was evaluated in a Der p 1-sensitization murine model. Results:, Mouse dendritic cells stimulated by L. plantarum triggered the release of interleukin-10 (IL-10), IL-12 p40, IL-12 p70 and tumor necrosis factor-alpha (TNF-,). IL-12 p40 secretion was dependent on nuclear factor-,B (NF-,B), mitogen-activated protein (MAP) kinases, Toll-like receptor 2 (TLR2), TLR9 and on the bacterial teichoic acid composition. Recombinant L. plantarum producing Der p 1 exhibited similar immunostimulatory properties as wild-type. Prophylactic intranasal pretreatment of mice with this recombinant strain prevented the development of the typical Th2-biased allergic response by a drastic reduction of specific IgE and the induction of protective allergen-specific IgG2a antibodies. Moreover, both wild-type or recombinant L. plantarum reduced airway eosinophilia following aerosolized allergen exposure and IL-5 secretion upon allergen restimulation. Conclusion:, By combining both Th1-type immunostimulatory properties and an efficient allergen delivery capacity, recombinant L. plantarum producing Der p 1 represents a promising vaccine against house dust mite allergy. [source]

    Comparative enzymology of native and recombinant house dust mite allergen Der p 1

    ALLERGY, Issue 3 2009
    J. Zhang
    Background:, The cysteine peptidase activity of group 1 house dust mite allergens is important for their allergenicity and may offer new therapeutic targets for allergy treatment. Hitherto, the design of specific inhibitors has been impeded because the availability of pure, fully active allergens has limited the implementation of drug screening campaigns. Similarly, investigation of the mechanisms by which peptidase allergens promote sensitization has also been restricted. Our aim was to compare the enzymology of recombinant and native forms of Der p 1 to establish if an easily expressed recombinant form of Der p 1 could be used as a drug discovery tool. Methods:, Enzymatic activity of natural and recombinant Der p 1 was compared fluorimetrically using a novel specific substrate (ADZ 50,059) and a novel specific active site titrant (ADZ 50,000). The effect of recombinant Der p 1 prodomain on the catalytic activity of both Der p 1 preparations was also examined. Results:, Although differing substantially in molecular weight, the enzymological properties of recombinant and native Der p 1 were indistinguishable. Our data show clearly by experiment that, in contrast to some suggestions, Der p 1 is not an enzyme of bifunctional mechanism. Conclusion:, The catalytic activity of Der p 1 is tolerant of glycosylation differences that occur at N150 when the protein is expressed in Pichia pastoris. This suggests that this recombinant protein may be suitable for drug design studies and in the elucidation of how peptidase activity promotes sensitization to peptidase and nonpeptidase bystander allergens. [source]

    The functional insufficiency of human CD4+CD25high T-regulatory cells in allergic asthma is subjected to TNF-, modulation

    ALLERGY, Issue 1 2008
    Y.-L. Lin
    Background:, Natural CD4+CD25highFoxp3+ regulatory T (nTreg) cells are important in maintaining immunologic tolerance, but their role in the pathogenesis of allergic asthma is unclear. We studied the function of nTreg cells in allergic asthmatic children and assessed the factors which may relate to the functional insufficiency of nTreg cells. Methods:, The percentage of CD4+CD25high Treg cells, the expression of Foxp3, and the cell-induced suppressive activity of nTreg cells isolated from nonatopic controls, allergic asthmatics, and allergen-specific immunotherapy (AIT)-treated asthmatic patients were studied. Results:, Although the percentage of nTreg in peripheral blood mononuclear cells was increased, the expression of Foxp3 and its cell-induced suppressive activity were significantly lower in Dermatophagoides pteronyssinus (Der p)-sensitive asthmatic children when compared to nonatopic controls. In contrast, the expression of Foxp3 and the functional activity of nTreg cells were reversed in allergic asthmatics who received AIT. The addition of recombinant tumor necrosis factor (TNF)-, directly downregulated Foxp3 expression and abrogated the cell-induced suppressive function of Treg cells. The anti-TNF-, reagent, etanercept, restored the functional activity and Foxp3 expression of CD4+CD25high Treg derived from allergic asthmatics. Conclusions:, The functional insufficiency of nTreg cells in patients with allergic asthma may be related to the enhanced production of TNF-, and its effect on the Foxp3 expression. These results may explain, in part, the effectiveness of anti-TNF-, therapy in the treatment of allergic asthma. [source]

    Induction of IL-10+ CD4+ CD25+ regulatory T cells with decreased NF-,B expression during immunotherapy

    PEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 1-Part-II 2010
    Yi-Giien Tsai
    Tsai Y-G, Chiou Y-L, Chien J-W, Wu H-P, Lin C-Y. Induction of IL-10+ CD4+ CD25+ regulatory T cells with decreased NF-,B expression during immunotherapy. Pediatr Allergy Immunol 2010: 21: e166,e173. © 2009 The Authors Journal compilation © 2009 Blackwell Munksgaard MyD88 is a major toll-like receptor (TLR) adaptor to activate NF-,B, which acts as a mater switch for allergic inflammation disease. Sterile hust dust extracts have been reported with TLR-dependent immunostimulatory activities. The aim of this study was to evaluate whether Dermatophagoides pteronyssinus (Der p) immunotherapy may increase IL-10+ CD4+ CD25+ T cells with modulating MyD88 signaling proteins, to decrease NF-,B expression. Peripheral blood mononuclear cells were isolated from patients before and after 1 yr of Der p immunotherapy, and also from matched control subjects. After 2 days of Der p-2 stimulation, intracellular IL-10 and Foxp3 expression of CD4+ CD25+ T cells were measured by flow-cytometry. The expression of IL-1 receptor-associated kinase (IRAK)-1 in cytoplasm and IFN-regulator factor-3 (IRF-3) with NF-,B/p65 in nuclei was determined by Western-blot analysis. Patients undergoing immunotherapy produced more soluble CD14, IL-10, and TGF-, that correlated with FEV1improvement (p < 0.05). In the immunotherapy group, the number of Foxp3+ CD4+ Treg cells increased more than the baseline status (25.06 ± 4.19 vs. 16.08 ± 3.54, p < 0.05). Additionally, increased IL-10 production with decreased IRAK-1 and NF-,B/p65 nuclear translocation was observed in sorted-purified Treg cells. IL-10+ CD4+ CD25+ Treg cells may respond to Der p-2 and down-regulate NF-,B/p65 expression to maintain immune tolerance during immunotherapy. [source]

    Crystallization and preliminary X-ray analysis of Der f 2, a potent allergen derived from the house dust mite (Dermatophagoides farinae)

    ACTA CRYSTALLOGRAPHICA SECTION D, Issue 6 2003
    Dana Roeber
    Although a number of allergens have been identified and isolated, the underlying molecular basis for the potent immune response is poorly understood. House dust mites (Dermatophagoides sp.) are ubiquitous contributors to atopy in developed countries. The rhinitis, dermatitis and asthma associated with allergic reactions to these arthropods are frequently caused by relatively small (125,129 amino acids) mite proteins of unknown biological function. Der,f,2, a major allergen from the mite D.,farinae, has been recombinantly expressed, characterized and crystallized. The crystals belong to the tetragonal space group I4122, with unit-cell parameters a = b = 95.2, c = 103.3,Å. An essentially complete (97.2%) data set has been collected to 2.4,Å at a synchrotron source. Attempts to solve the crystal structure of Der,f,2 by molecular replacement using the NMR coordinates for either Der,f,2 or Der,p,2 (the homologous protein from D.,pteronyssinus) failed, but preliminary searches using the crystalline Der p 2 atomic coordinates appear to be promising. [source]

    Surfactant protein D inhibits mite-induced alveolar macrophage and dendritic cell activations through TLR signalling and DC-SIGN expression

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 1 2010
    C-F Liu
    Summary Background Surfactant protein D (SP-D), a secreted pattern recognition molecule associated with pulmonary innate immunity, has been shown to mediate the clearance of pathogens in multiple ways. However, how SP-D interacts with alveolar macrophages (AMs) and dendritic cells (DCs) during allergen exposure remains unclear. Objective This study was performed to characterize the immunomodulatory effects of SP-D on mite allergen (Dermatophagoides pteronyssinus, Der p)-induced inflammatory signalling in AMs and DCs. Methods Murine AM, alveolar macrophage cell line derived from BALB/c mice (MH-S cells), and human monocyte-derived dendritic cells (MDDC) were used as model systems. The production of nitric oxide (NO) and TNF-,, expression of surface Toll-like receptors (TLRs), and expression of the C-type lectin receptor known as dendritic cell (DC)-specific ICAM-grabbing non-integrin (DC-SIGN) were measured as a function of pretreatment with SP-D and subsequent exposure to Der p. Der p-dependent cellular activations that were modified by SP-D in these model systems were then identified. Results Pretreatment of MH-S cells with SP-D reduced Der p-dependent production of NO, TNF-,, and the downstream activations of IL-1 receptor-associated kinase, mitogen activated protein kinase (MAPK) kinase, and nuclear factor-,B. SP-D interacted with CD14 such that CD14 binding to Der p was inhibited and Der p-induced signalling via TLRs was blocked. DC-SIGN expression was suppressed by Der p in MH-S and MDDC; this down-regulation of DC-SIGN expression was prevented by pretreatment with SP-D. Conclusions These results indicated that the inhibition of Der p-induced activation of MH-S and MDDC by SP-D is mediated through suppression of the CD14/TLR signalling pathway and maintenance of DC-SIGN expression, which may protect allergen-induced airway inflammation. Cite this as: C-F Liu, M. Rivere, H-J Huang, G. Puzo and J-Y Wang, Clinical & Experimental Allergy, 2010 (40) 111,122. [source]

    Production of native and modified recombinant Der p 1 molecules in tobacco plants

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 5 2009
    D. Burtin
    Summary Background As a complex molecule requiring post-translational processing, it has been difficult to produce the Der p 1 major allergen from the Dermatophagoides pteronyssinus house dust mite in a recombinant form. Objective Here, we tested whether transgenic tobacco plants are suitable to express Der p 1, either as a wild-type molecule or as variants lacking N -glycosylation sites (Gly,) and/or cysteine protease activity (Enz,). Methods Using Agrobacterium tumefaciens -based transformation, pro Der p 1 molecules bearing mutations within either the N -glycosylation sites (N34Q, N150Q) and/or the cysteine protease-active site (C132V) were expressed in tobacco plants. After purification by ion exchange chromatography, allergens were characterized using immunoblotting, circular dichroism (CD), as well as basophil and T lymphocyte stimulation assays. Results Four forms of recombinant Der p 1 (i.e. wild-type Gly+/Enz+, as well as Gly,/Enz+, Gly+/Enz, or Gly,/Enz, variants) were successfully expressed in tobacco leaves as pro Der p 1 molecules. Spontaneous cleavage of the pro-peptide was observed in tobacco leaf extracts for all forms of recombinant Der p 1 (r Der p 1). CD confirmed that all r Der p 1 molecules, with the exception of the Gly,/Enz, variant, exhibited secondary structures comparable to the natural protein. A cysteine protease activity was associated only with the Gly+/Enz+ form. All these molecules exhibit a profile similar to natural Der p 1 with respect to IgE immunoreactivity, basophil activation and T cell recognition. Conclusion A tobacco plant expression system allows the production of various forms of mature Der p 1, which could be used for diagnostic or immunotherapeutic purposes. [source]

    An immunoglobulin E-reactive chimeric human immunoglobulin G1 anti-idiotype inhibits basophil degranulation through cross-linking of Fc,RI with Fc,RIIb

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 2 2008
    S. J. Wigginton
    Summary Background IgE binds to mast cells and basophils via its high-affinity receptor, Fc,RI, and cross-linking of Fc,RI-bound IgE molecules by allergen leads to the release of allergic mediators characteristic of type I hypersensitivity reactions. Previous work has shown that cross-linking of Fc,RI with Fc,RIIb, an ITIM-containing IgG receptor, leads to inhibition of basophil triggering. 2G10, a chimeric human IgG1 anti-idiotype, has broad reactivity with human IgE and as such has the potential to bind simultaneously to Fc,RI-bound IgE, via its Fab regions, and the negative regulatory receptor, Fc,RIIb, via its Fc region. Objective To assess the ability of human 2G10 to inhibit anti-IgE and allergen-driven basophil degranulation through cross-linking of Fc,RI-bound IgE with Fc,RIIb. Methods 2G10 was assessed for its ability to bind to Fc,RIIb on transfected cells and on purified basophils. In the basophil degranulation assay, basophils were purified from peripheral blood of atopic individuals and activated with either anti-IgE or the house dust mite allergen Der p 1, in the presence or absence of human 2G10. Basophil activation was quantified by analysis of CD63 and CD203c expression on the cell surface, and IL-4 expression intracellularly, using flow cytometery. Results Human 2G10 was able to bind to Fc,RIIb on transfected cells and on purified basophils, and induce a dose-dependent inhibition of both anti-IgE and Der p 1-driven degranulation of basophils. Conclusion The inhibition of basophil degranulation by the human IgG1 anti-idiotype 2G10 highlights the therapeutic potential of IgE-reactive IgG antibodies in restoring basophil integrity through recruitment of the inhibitory receptor Fc,RIIb. [source]

    Influence of short-term exposure to airborne Der p 1 and volatile organic compounds on skin barrier function and dermal blood flow in patients with atopic eczema and healthy individuals

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 3 2006
    J. Huss-Marp
    Summary Background Epidemiological studies indicate environmental pollutants to be involved in the increase in the prevalence of allergic diseases. In human exposure studies, volatile organic compounds (VOCs) have been shown to cause exacerbations of allergic asthma whereas, no data concerning atopic eczema (AE) are available. Objective We investigated the effect of airborne VOCs on the skin of patients with AE and controls in the presence or absence of house dust mite allergen, Der p 1. Methods In a double-blind crossover study, 12 adults with AE and 12 matched healthy volunteers were exposed on their forearms to Der p 1 and subsequently to a mixture of 22 VOCs (M22, 5 mg/m3) in a total body exposure chamber for 4 h. Transepidermal water loss (TEWL) and skin blood flow were measured in all subjects before, during and after exposure. Additionally, an atopy patch test (APT) with Der p 1 was applied to the skin after exposure. Results A significant increase in transepidermal water loss was observed 48 h after exposure to VOCs as compared with exposure with filtered air in all individuals (mean difference: +34%; 95% Confidence Interval: 7,69%). Prior Der p 1 exposure resulted in a significant rise of dermal blood flow after 48 h in patients with AE but not in controls. Six out of seven patients showed enhanced atopy patch test (APT) reactions to HDM allergen after previous exposure to VOCs. Conclusion Our results show that exposure to VOCs , at concentrations commonly found in indoor environments , can damage the epidermal barrier and enhance the adverse effect of Der p 1 on sensitized subjects with AE. These findings may contribute to a better understanding of the mechanisms underlying the increase in prevalence and exacerbation of AE. [source]

    Transgenic mice expressing the T cell antigen receptor specific for an immunodominant epitope of a major allergen of house dust mite develop an asthmatic phenotype on exposure of the airways to allergen

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 7 2005
    E. R. Jarman
    Summary Background Current studies on mechanisms underlying allergen-induced pulmonary inflammation and asthma are hampered by the lack of appropriate physiological in vivo models that reflect the natural route of allergen exposure and sensitization. Objective To generate and phenotype a transgenic mouse strain expressing the T cell receptor (TCR) specific for an immunodominant domain of the major inhalant allergen Dermatophagoides pteronyssinus species of house dust mite (Der p 1), for the development of an in vivo model of allergic asthma. Methods Der p 1 transgenic mice were generated using TCR-,, derived from a CD4+ T cell hybridoma reactive with Der p 1 residues p 110,131. The frequency and functional activity of peripheral T cells were determined and parameters of airway inflammation assessed following allergen challenge of the airways with Der p 1. Results CD4+ T cells are functionally active, exhibiting dose-dependent proliferation and IL-4 production on primary stimulation with Der p 1 or Der p 1, p 110,131 in vitro, independent of in vivo antigen priming. On sensitization of the airways with allergen, in the absence of systemic priming or the application of adjuvants, the TCR transgenic mice develop airway inflammation characterized by a marked lymphocytic and eosinophilic infiltrate with goblet cell hyperplasia and enhanced mucin production. Conclusion The Der p 1 TCR transgenic mice provide a model for investigating the pathophysiological mechanisms of pulmonary inflammation following sensitization by exposure of the airways to allergen and for investigating the mode of action and efficacy of novel immunotherapeutics. [source]

    Anaphylaxis after hamster bites , identification of a novel allergen

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 7 2004
    D. L. Lim
    Summary Background Hamsters are popular household pets and anaphylaxis after their bites have described. However, the putative allergen has not been identified. Objective This study was conducted to identify the allergen causing dwarf hamster (Phodopus sungoris) bite-induced anaphylaxis. Methods Two children with hamster bite-induced anaphylaxis were enrolled. They both had negative results to skin testing and specific IgE to hamster epithelium. However, they were both allergic to Dermatophagoides pteronyssinus (Der p). Identification of the putative IgE-binding allergens from the hamster saliva was performed using immunoblot analysis. Results A specific IgE-binding component at 21 kD in the hamster saliva was identified. ELISA inhibition tests showed partial inhibition with Der p. Conclusions The putative allergen from the hamster saliva causing dwarf hamster-induced anaphylaxis was identified. Possible cross-reactivity with Der p was demonstrated. Further studies will be needed to identify the exact nature and function of this allergen. [source]

    Allergens, Der p 1, Der f 1, Fel d 1 and Can f 1, in newly bought mattresses for infants

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 11 2002
    R. De Boer
    Summary Background To avoid allergen exposure of newborn babies, the use of a new mattress for the baby bed may be recommended. However, it is not certain that new mattresses are always free of allergens. Objective In the present study the allergen content of new infant mattresses was investigated. Methods Dust samples were vacuumed from 90 new mattresses for infant beds bought in 50 different Dutch shops, and the concentrations of Der p 1, Der f 1, Fel d 1 and Can f 1 were determined by radioimmunoassays. Results Most mattresses contained some allergen and often the allergen concentrations were surprisingly high. Only 15 of the 90 mattresses contained no detectable amounts of any of the four allergens. The highest concentration found for each allergen was 3.1, 46.5, 20.2 and 95.7 µg/g of dust, respectively. However, the total amount of allergen in a mattress was still rather low because the new mattresses contained only modest amounts of dust. Baby mattresses more often contained an increased allergen load than the larger, standard-sized, infant mattresses. This may be caused by differences in manufacturing procedure. Also, mattresses that were sold without a plastic encasement more often contained an increased allergen load. Conclusions It is advisable to buy a mattress that is wrapped in plastic, but it may still contain a substantial amount of allergen. Thorough vacuuming of a newly bought mattress before it is installed on a child's bed, is also advisable. After instalment, regular vacuuming of the mattress and washing of the bed linen as well as measures to eliminate allergen reservoirs in other parts of the house are important, because our observations indicate that mattresses easily pick up allergens from the environment. [source]

    Stimulatory and inhibitory epitopes in the T cell responses of mice to Der p 1

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 6 2002
    A. G. Jarnicki
    Summary Background The responses of mice to the mite allergen Der p 1 have been used to study the mechanisms of allergic sensitization and the development of new types of immunotherapy. Many of the studies require a knowledge of the T cell epitopes, and because Der p 1 is polymorphic, the effect of natural amino acid substitution in the allergen. The intranasal administration of peptides containing T cell epitopes can induce a mucosal tolerance but it is not known if the major activity is limited to stimulatory peptides and if, as found for autoimmunity, some epitopes are not inhibitory. Objective To determine and compare the sequences of Der p 1 which contain stimulatory epitopes for the high responding H-2b and H-2q mice and the sequences which induce tolerance by intranasal administration of peptides. Methods T cell responses of mice immunized with Der p 1 were measured by in vitro T cell stimulation assays so an extensive study of epitope recognition and intranasal tolerance could be made. Synthetic peptides were used to examine the stimulatory and inhibitory ability of all Der p 1 sequences and to map the major H-2b epitope in detail. This included the effect of the common polymorphic amino acid 124 substitution found within this epitope. Results Three and two regions, respectively, were found to contain stimulatory T cell epitopes for H-2b and H-2q mice. The peptides in these regions were also the most active at inducing intranasal tolerance for the responding haplotype. The correspondence between inhibitory and stimulatory peptides was maintained for the fine mapping of the major H-2b epitope. This was found about a core region of 118,126 which was overlapping but separate to a consensus sequence for the binding of endogeneous peptides. Peptides with alanine at the naturally polymorphic residue 124 stimulated and inhibited responses to Der p 1 more effectively, while peptides with the valine 124 variant were immunogenic but poorly cross-reactive. Conclusions The intranasal administration of peptides representing each of five epitopes recognized by two strains of mice were able to induce mucosal tolerance and the major tolerizing activity was limited to these epitopes. The position of the core major epitope for C57 mice, which differs from a previously predicted epitope, and its specificity for the natural alanine 124 variant is described. [source]

    Biochemical and immunological characterization of a recombinant precursor form of the house dust mite allergen Der p 1 produced by Drosophila cells

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 5 2000
    Jacquet
    Background The major house dust mite allergen Der p 1 elicits strong IgE antibody responses in patients suffering from mite allergy. Objective This study reports the expression and characterization of a recombinant precursor form of Der p 1 secreted as ProDer p 1 from insect cells. Methods The cDNA coding for ProDer p 1 was cloned downstream to the gp67 signal peptide, starting from commercial cDNA encoding Der p 1 and PCR-amplified ProDer p 1 genomic fragment. ProDer p 1, expressed in Drosophila cells and purified from culture medium, was compared to Der p 1 isolated from mite culture, in terms of glycosylation, enzymatic activity as well as IgG- and IgE-binding capacity. Results Sequence analysis of the genomic clone of ProDer p 1 revealed that, besides two introns in the mature Der p 1 coding sequence, two introns were also present in the propeptide coding sequence. ProDer p 1 was purifed to homogeneity by a combination of ion-exchange, hydroxyapatite and gel filtration chromatographies. The precursor form of Der p 1 could be processed in vitro into mature Der p 1 under acidic and reducing conditions. Carbohydrate analysis clearly indicated that ProDer p 1 expressed from insect cells was glycosylated and that glycan structures were located only in the prosequence. ProDer p 1 displayed a similar immunoreactivity towards IgE, monoclonal and polyclonal IgG antibodies compared to natural Der p 1. Specific activity measurements using synthetic substrates clearly indicated that, contrary to natural Der p 1, ProDer p 1 was totally enzymatically inactive. Conclusions The expression of an enzymatically inactive and highly antigenic ProDer p 1 zymogen molecule could be a suitable strategy for the development of in vitro diagnosis test as well as for specific immunotherapy. [source]

    HLA DPB1*0201 allele is negatively associated with immunoglobulin E responsiveness specific for house dust mite allergens in Taiwan

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 4 2000

    Background House dust mite (HDM) Dermatophagoides pteronyssinus is the most important source of indoor allergens that cause allergic diseases in Taiwan. We prepared purified HDM allergens (Der p 1, Der p 2 and Der p 5) to detect allergen-specific immunoglobulin (Ig) E responsiveness among a large number of test subjects. The robust genetic typing system for HLA class II genes also facilitated the study on association of HLA and allergic response toward HDM. Objective This study intended to investigate the association between HLA class II alleles and the IgE responsiveness to the major allergens from HDM, D. pteronyssinus. Methods Two hundred and forty-eight subjects were selected for HLA association study. Plasma HDM allergen (Der p 1, Der p 2, Der p 5) -specific IgE and Der p 2-specific IgG antibodies were detected by ELISA, while HLA class II -DRB1, -DQA1, -DQB1, -DPB1 genetic polymorphism was determined by polymerase chain reaction/sequence-specific oligonucleotide probe hybridization (PCR/SSOPH). Statistical comparison of the allelic distribution of each HLA class II genes among the individuals with/without HDM allergen-specific IgE and IgG antibodies were performed. Results There was no significant association between HLA DRB1, DQB1, DQA1 alleles and HDM-specific IgE responsiveness noted. Only DRB1*0803 and the linked DQA1*0103 alleles showed positive association with Der p 5-specific IgE responsiveness. However, we found that HLA-DPB1*1301 predisposed subjects to IgE responsiveness to HDM Der p 5. HLA DPB1*0501 was weakly associated with the IgE responsiveness to HDM Der p 1 and Der p 5. There was a strong negative association between the HLA-DPB1*0201 allele with IgE responsiveness to Der p 1 (OR: 0.30, P , 0.0001, P , 0.0007, Pc , 0.010). Conclusion We clearly observed the association between HLA DPB1 alleles and specific IgE responsiveness to HDM major allergens. The molecular mechanism of HLA-DPB1*0201 involvement in protecting subjects from HDM-specific IgE responsiveness awaits further investigation. [source]

    Total and specific IgE (house dust mite and intestinal helminths) in asthmatics and controls from Gondar, Ethiopia

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 3 2000
    Selassie
    Background The role, if any, of parasitosis in the development of asthma remains incompletely understood; both ,protective' and ,predictive' associations have been reported. We report a study which examined immunoglobulin (Ig) E responses to two common helminths in asthmatics living in Ethiopia. Objective To compare the frequencies of specific IgE antibodies to Ascaris and Necator species and to Der p 1 among 84 adult asthmatics and a referent group of nonasthmatics. Methods A case-control analysis. Results Total IgE levels were not different between the two groups. The presence of specific IgE to Der p 1 was strongly associated with asthma (P = 0.001). Raised levels of Ascaris-(P = 0.010) and Necator- (P = 0.001) specific IgE antibodies were more common among referents; there were no associations between specific IgE production to Der p 1 and either of the two parasites. Conclusion These findings confirm the association between Der p 1 sensitization and asthma among urban, adult Ethiopians. While they also indicate a negative relationship with two indicators of helminth infestation it appears that this is not mediated through the immunological response to common aeroallergens. [source]

    Airway eosinophilia is not a requirement for allergen-induced airway hyperresponsiveness

    CLINICAL & EXPERIMENTAL ALLERGY, Issue 1 2000
    Tournoy
    Background House dust mites (HDMs) are the major source of perennial allergens causing human allergic asthma. Animal models mimicking as closely as possible the allergic features observed in human asthma are therefore interesting tools for studying the immunological and pathophysiological mechanisms involved. Especially the role of eosinophils and allergen-specific immunoglobulin (Ig) E in the pathophysiology of airway hyperresponsiveness (AHR) remains a subject of intense debate. Objective To develop a mouse model of allergic airway inflammation and hyperresponsiveness based on the use of purified house dust mite allergen (Der p 1) as clinical relevant allergen. Furthermore, we studied the effects of low dose allergen exposure on the airway eosinophilia and AHR. Methods On day 0, C57Bl/6 mice were immunized with purified Der p 1 intraperitoneally. From day 14,20, the mice were exposed daily to a 30-min aerosol of different concentrations of house dust mite extract. Results Mice, actively immunized with Der p 1 and subsequently exposed to HDM aerosols, developed AHR, eosinophil infiltration of the airways and allergen-specific IgE. Moreover, lowering the concentration of the HDM aerosol also induced AHR and IgE without apparent eosinophil influx into the airways. Der p 1-sensitized mice exposed to PBS produced IgE, but did not show AHR or eosinophil influx. Conclusion This in vivo model of HDM-induced allergic airway changes suggests that AHR is not related to either eosinophil influx or allergen-specific serum IgE, thereby reducing the importance of these factors as essential elements for allergic AHR. [source]