Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A(2A) adenosine receptor (A(2A)AR) has potent anti-inflammatory properties, which may be important in the regulation of airway reactivity and inflammation. Inflammatory cells that possess A(2A)AR also produce nitrosative stress, which is associated with pathophysiology of asthma, so we hypothesized that A(2A)AR deficiency may lead to increased airway reactivity and inflammation through nitrosative stress. Thus the present study was carried out to investigate the role of A(2A)AR on airway reactivity, inflammation, NF-kappaB signaling, and nitrosative stress in A(2A)AR knockout (KO) and wild-type (WT) mice using our murine model of asthma. Animals were sensitized intraperitoneally on days 1 and 6 with 200 microg of ragweed, followed by aerosolized challenges with 0.5% ragweed on days 11, 12, and 13, twice a day. On day 14, airway reactivity to methacholine was assessed as enhanced pause (Penh) using whole body plethysmography followed by bronchoalveolar lavage (BAL) and lung collection for various analyses. Allergen challenge caused a significant decrease in expression of A(2A)AR in A(2A) WT sensitized mice, with A(2A)AR expression being undetected in A(2A) KO sensitized group leading to decreased lung cAMP levels in both groups. A(2A)AR deletion/downregulation led to an increase in Penh to methacholine and influx of total cells, eosinophils, lymphocytes, and neutrophils in BAL with highest values in A(2A) KO sensitized group. A(2A) KO sensitized group further had increased NF-kappaB expression and nitrosative stress compared with WT sensitized group. These data suggest that A(2A)AR deficiency leads to airway inflammation and airway hyperresponsiveness, possibly via involvement of nitrosative stress in this model of asthma.
Am J Physiol Lung Cell Mol Physiol 2007 Jun
PMID:Enhanced airway reactivity and inflammation in A2A adenosine receptor-deficient allergic mice. 1729 74

Over the last few decades, the prevalence of allergic diseases has increased dramatically in developed nations. The resulting burden on health care systems worldwide has provoked a whole series of research initiatives among allergy experts and commercial companies that aim to develop novel tests to improve the diagnostic risk assessment and early preventive treatment of the disease. The advent of protein microarray technology has inspired the development of miniaturized immunological applications that permit the simultaneous analysis of huge numbers of disease-related parameters. Allergen microarrays have been developed for the monitoring of patient-specific antibody profiles to a previously unknown variety of allergens in a single analytical step. This has been accomplished by the successful adaptation of solid-phase antibody assays for the detection of surface-bound allergens to the microarray format, the development of appropriate assay conditions, and the improvement of software-guided microarray image analysis. Here we report a protocol for the development and analysis of food allergen microarrays.
Methods Mol Biol 2007
PMID:Allergen microarrays for the diagnosis of specific IgE against components of cow's milk and hen's egg in a multiplex biochip-based immunoassay. 1836 10

Arachidonate 15-lipoxygenase (LO)-1 has been implicated in allergic inflammation and asthma. The overall effect of 15-LO in allergic inflammation in vivo is, however, unclear. This study investigates systemic allergen sensitization and local allergic airway inflammation and remodeling in mice lacking the murine 12/15-LO, the ortholog to human 15-LO-1. Upon systemic sensitization with intraperitoneal ovalbumin, 12/15-LO-/- mice produced elevated levels of allergen-specific immunoglobulin E compared with wild-type (Wt) controls. However, when challenged with repeated aerosolized allergen, sensitized 12/15-LO-/- mice had an impaired development of airway allergic inflammation compared with Wt controls, as indicated by reduced bronchoalveolar lavage fluid leukocytes (eosinophils, lymphocytes, macrophages) and Th2 cytokines (IL-4, IL-5, IL-13), as well as tissue eosinophils. Allergen-induced airway epithelial proliferation was also significantly attenuated in 12/15-LO-/- mice, whereas goblet cell hyperplasia was unaffected. However, 12/15-LO-/- mice had significantly reduced luminal mucus secretions compared with Wt controls. The repeated allergen challenges resulted in a dramatic increase of alpha-smooth muscle actin-positive alveolar cells in the peripheral airways, a phenomenon that was significantly less developed in 12/15-LO-/- mice. In conclusion, our data suggest that 12/15-LO-/- mice, although having a fully developed systemic sensitization, did not establish a fully developed allergic airway inflammation and associated manifestations of central and peripheral airway remodeling. These data suggest that 12/15-LO-derived metabolites play an important pathophysiologic role in allergen-induced inflammation and remodeling. Hence, pharmacologic targeting of the human 15-LO-1 may represent an attractive therapeutic strategy to control inflammation and remodeling in asthma.
Am J Respir Cell Mol Biol 2008 Dec
PMID:Mice lacking 12/15-lipoxygenase have attenuated airway allergic inflammation and remodeling. 1851 9

Allergen-specific T-cell lines established from allergic patients provide the opportunity of investigating T-cell functions at the poly- or oligoclonal level. T-cell lines are useful in determining the presence or absence of antigen-specific T-cell reactivity. However, to obtain detailed knowledge of the action of T cells with clearly defined features, for example epitope specificity or phenotype, T-cell clones are necessary.The frequency of allergen-specific T cells in peripheral blood mononuclear cells (PBMC) tends to be low and so stimulation of PBMC with single allergens often results in low allergen-specific reactivity or requires high doses of the allergen. In contrast, the stimulation of PBMC with whole allergen extract results in stronger reactivity because a greater spectrum of T-cell specificities is addressed. Therefore, for the investigation of polyclonal reactivity toward single allergens it is useful to establish T-cell lines, which represent an allergen-specific enrichment of T cells from the respective individual. These T cells are poly- or oligoclonal and might possess different epitope specificities. The method described here is based on experiences with human T-cell lines and clones specific for several allergens from grass pollens and tree pollens.
Methods Mol Med 2008
PMID:Production of T-cell lines. 1861 2

Monoclonal antibodies (mabs) are powerful tools for the quantification, detection, and targeting of specific molecules. Allergen-specific mabs are important for the quantification of major allergens in allergen preparations used for allergen-specific immunotherapy and allergy diagnosis. Indeed, progress in the understanding of the mechanisms of the immunological responses underlying allergic disease would not have been possible without the use of mabs. Quantification assays are also important in the assessment of environmental allergen exposure and monitoring of avoidance procedures.Mabs against human IgE provide the basis for various test systems for the detection of specific and nonspecific IgE. Mabs raised against IgE or defined cytokines or cytokine receptors have potential as neutralizing reagents in vivo for the treatment of allergic diseases.Allergen-specific mabs are also valuable tools for the localization of allergens within their source material and the characterization of allergens derived from natural sources and by recombinant technologies. Furthermore they are often used for the isolation of allergens from complex extracts by affinity chromatography. The procedure described in this chapter has been used successfully to produce mabs against numerous allergens from house dust mites, insect venoms, cat, hens egg white, tree-, grass-, and herb pollens, and fungi, with the ultimate aim of obtaining matched antibody pairs to establish two-site binding assays for the quantification of major allergens. The method has also been used successfully to generate mabs against human IgE.
Methods Mol Med 2008
PMID:Monoclonal antibodies. 1861 8

Specific allergen immunotherapy is an effective treatment for IgE-mediated allergic disease and involves T- and B-cell mediated events. IgE receptors on the surface of antigen-presenting cells facilitate the presentation of allergens in the presence of specific IgE antibody resulting in T-cell activation. Interference with these IgE-dependent mechanisms by 'blocking' IgG antibodies may downregulate T-cell responses and manifest as a reduction in allergic responses in vivo. The vigor of proliferative responses by T-cell clones is representative of the binding of allergen-IgE complexes to B cells. Therefore, a simplified assay can be employed that measures the binding of allergen-IgE complexes to B cells instead of a more complex assay involving proliferative assays using antigen-specific T-cell clones. Allergen-IgE complexes can be easily detected by flow cytometry and this simplified technique is called the IgE-facilitated allergen binding (IgE-FAB) assay which is described in this chapter.
Methods Mol Med 2008
PMID:The facilitated antigen binding (FAB) assay--a protocol to measure allergen-specific inhibitory antibody activity. 1861 14

Allergen isoforms can differ in their IgE and T cell recognition patterns, and thus might have an impact on the selection of candidates for molecule-based diagnostic and therapeutic approaches. The present study aimed at the identification and characterization of isoforms of Art v 1, the mugwort pollen major allergen. In addition, single Art v 1 domains were physicochemically and immunologically characterized. For this purpose, the Art v 1 cDNA was radiolabeled and used to screen a mugwort pollen cDNA library. Positive clones were sequenced and used for the production of recombinant proteins in Escherichia coli using the pHIS-Parallel2 vector. Protein purification was performed by affinity- and ion exchange chromatography. Antibody binding to the recombinant proteins was determined by immunoblot, ELISA, cross-inhibition experiments, and mediator release assays. We could identify 7 Art v 1 isoforms differing in 1-6 amino acid residues. Interestingly, all amino acid variations were restricted to the proline domain carrying the molecule's post-translational modifications. No significant difference in IgG or IgE reactivity could be observed between Art v 1 isoforms and the defensin domain produced in E. coli. When expressed in E. coli, the proline domain was not recognized by Art v 1-specific antibodies. Our results demonstrated that the relevant IgE epitopes of Art v 1 are located on the defensin domain and suggest the involvement of carbohydrates in the allergenicity of natural Art v 1. Plant-based expression systems could help to reveal possibly different glycosylation patterns and IgE binding properties of Art v 1 isoforms. These findings have direct implications on the development of novel tools for mugwort pollen allergy diagnosis and therapy.
Mol Immunol 2009 Jan
PMID:Immune recognition of novel isoforms and domains of the mugwort pollen major allergen Art v 1. 1905 64

Although the role of arachidonic acid (AA) metabolism to eicosanoids has been well established in allergy and asthma, recent studies in neoplastic cells have revealed that AA remodeling through phospholipids impacts cell survival. This study tests the hypothesis that regulation of AA/phospholipid-remodeling enzymes, cytosolic phospholipase A(2) alpha(cPLA(2)-alpha, gIValphaPLA(2)) and CoA-independent transacylase (CoA-IT), provides a mechanism for altered eosinophil survival during allergic asthma. In vitro incubation of human eosinophils (from donors without asthma) with IL-5 markedly increased cell survival, induced gIValphaPLA(2) phosphorylation, and increased both gIValphaPLA(2) and CoA-IT activity. Furthermore, treatment of eosinophils with nonselective (ET18-O-CH(3)) and selective (SK&F 98625) inhibitors of CoA-IT triggered apoptosis, measured by changes in morphology, membrane phosphatidylserine exposure, and caspase activation, completely reversing IL-5-induced eosinophil survival. To determine if similar activation occurs in vivo, human blood eosinophils were isolated from either normal individuals at baseline or from subjects with mild asthma, at both baseline and 24 hours after inhaled allergen challenge. Allergen challenge of subjects with allergic asthma induced a marked increase in cPLA(2) phosphorylation, augmented gIValphaPLA(2) activity, and increased CoA-IT activity. These findings indicate that both in vitro and in vivo challenge of eosinophils activated gIValphaPLA(2) and CoA-IT, which may play a key role in enhanced eosinophil survival.
Am J Respir Cell Mol Biol 2009 Sep
PMID:Regulation of arachidonate remodeling enzymes impacts eosinophil survival during allergic asthma. 1915 22

An increasing number of patients are suffering from allergic diseases such as rhinoconjunctivitis, atopic eczema, uticaria, anaphylaxis, and food and drug allergies. Although it is possible to measure a multitude of allergen-specific IgE antibodies by radio or enzyme immunoassays in the patients' blood, these tests are expensive, time-consuming, and usually need a rather high volume of reagent solutions (allergens and blood). Protein microarrays offer the possibility to circumvent these limitations. The described in vitro allergy testing system is based on microscopic glass slides activated with glycidyloxypropyl-trimethoxysilane. Allergen solutions (allergen extracts and/or purified allergens; approximately 10 nL) are printed on the activated glass surface with a piezoelectric spotting machine. The protein components of the allergen solutions are immobilized on the modified glass surface via hydrophobic interaction and/ or covalent binding. After a blocking step, the slides are incubated with the respective diluted serum sample (approximately 25 microL serum required) and bound IgE antibodies are detected with a secondary horseradish peroxidase (HRP) labelled anti-human-IgE antibody via chemiluminescence. The measurement can be performed automatically with the so called PASA system. Test results are directly visualized with a CCD-camera. Analytical and clinical data have shown that the microarray-based test format offers significant advantages in time and costs compared with traditional test formats. The described allergen microarray demonstrated a sufficient qualitative reproducibility and enabled the distinction between allergic and non-allergic patients. Detection limits of 0.35 kU/L (r Bet v1), 0.16 kU/L (PLA2), 1.9 kU/L (Der p1), and 41 kU/L (total IgE) were achieved.
Methods Mol Biol 2009
PMID:Detection of known allergen-specific IgE antibodies by immunological methods. 1921 17

Art v 3, the lipid-transfer protein (LTP) of Artemisia vulgaris pollen is a relevant allergen showing frequent cross-reactivity with homologues in other plants. Here we report the identification of four full-length Art v 3 sequences obtained by cDNA cloning using mass spectrometry-based sequencing. Two isoforms, Art v 3.0201 and Art v 3.0301 were expressed as soluble proteins in Escherichia coli Rosetta-gami B(DE3) pLysS using different expression systems. Purified natural and recombinant Art v 3 demonstrated similar secondary structures in circular dichroism analysis. All preparations showed high thermal stability but low resistance to gastric digestion with pepsin. Patient-specific IgE reactivity patterns to natural or recombinant isoallergens were observed among Art v 3-sensitized subjects. Using Immuno Solid-phase Allergen Chip (ISAC) assays, frequent cross-reactivity of Art v 3 with LTPs from peach and hazelnut was shown. The biological activity of both isoforms was comparable to the natural allergen in basophil release assays. The newly identified sequences provide the basis for recombinant mugwort LTP production enabling batch-to-batch reproducibility and thus ensuring high-quality products for diagnosis and therapy.
Mol Immunol 2009 Jun
PMID:Isoform identification and characterization of Art v 3, the lipid-transfer protein of mugwort pollen. 1940 80


<< Previous 1 2 3 4 5 6 7 8 9 Next >>