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Articles by Andrea Harrer in JoVE
JoVE Immunology and Infection
Basophil Aktivering test för utredning av IgE-medierade mekanismer i Läkemedelsöverkänslighet
1Department of Molecular Biology, University of Salzburg, 2Department of Neurology, Paracelsus Medical University, 3Department of Dermatology, Paracelsus Medical University, 4Bühlmann Laboratories, 5Christian Doppler Laboratory for Allergy Diagnosis and Therapy, University of Salzburg
Basophil aktivering test är ett kraftfullt verktyg för detektering av IgE-beroende allergier
Other articles by Andrea Harrer on PubMed
Isoform Identification and Characterization of Art V 3, the Lipid-transfer Protein of Mugwort Pollen
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.
Characterization of Plant Food Allergens: an Overview on Physicochemical and Immunological Techniques
Allergy to plant-derived foods is a highly complex disorder with clinical manifestations ranging from mild oral, gastrointestinal, and cutaneous symptoms to life-threatening systemic conditions. This heterogeneity in clinical manifestations has been attributed to different properties of allergenic molecules. Based on this fact, symptom elicitors were grouped into class I and pollinosis-associated class II food allergens, but clear distinction is rather ambiguous. Moreover, mechanisms underlying food sensitization are not fully understood yet, and food allergy management most often relies on patient's compliance to avoid suspected foods. Therefore, recent efforts aim at the investigation of plant food allergies at the molecular level. This review provides an overview on currently available techniques for allergen characterization and discusses their application for investigation of plant food allergens. Data obtained by an array of physicochemical analyses, such as allergen structure, integrity, aggregation, and stability, need to be linked to results from immunological methods at the level of IgE and T-cell reactivity. Such knowledge allows the development of computational algorithms to predict allergenicity of novel foods being introduced by biotechnological industry. Furthermore, molecular characterization is an indispensable tool for molecule-based diagnosis and future development of safer patient-tailored specific immunotherapy in plant food allergy.
Diclofenac Hypersensitivity: Antibody Responses to the Parent Drug and Relevant Metabolites
Hypersensitivity reactions against nonsteroidal antiinflammatory drugs (NSAIDs) like diclofenac (DF) can manifest as Type I-like allergic reactions including systemic anaphylaxis. However, except for isolated case studies experimental evidence for an IgE-mediated pathomechanism of DF hypersensitivity is lacking. In this study we aimed to investigate the possible involvement of drug- and/or metabolite-specific antibodies in selective DF hypersensitivity.
Molecular Characterization of Api G 2, a Novel Allergenic Member of the Lipid-transfer Protein 1 Family from Celery Stalks
Scope: Celery represents a relevant cross-reactive food allergen source in the adult population. As the currently known allergens are not typical elicitors of severe symptoms, we aimed to identify and characterize a non-specific lipid transfer protein (nsLTP).Methods and results: MS and cDNA cloning were applied to obtain the full-length sequence of a novel allergenic nsLTP from celery stalks. The purified natural molecule consisted of a single isoallergen designated as Api g 2.0101, which was recombinantly produced in Escherichia coli Rosetta-gami. The natural and recombinant molecules displayed equivalent physicochemical and immunological properties. Circular dichroism revealed a typical α-helical fold and high thermal stability. Moreover, Api g 2 was highly resistant to simulated gastrointestinal digestion. As assessed by ELISA, thermal denaturation did not affect the IgE binding of Api g 2. Natural and recombinant Api g 2 showed similar allergenic activity in mediator release assays. Api g 2-specific IgE antibodies cross-reacted with peach and mugwort pollen nsLTPs.Conclusion: Based on our results, it can be anticipated that inclusion of recombinant Api g 2 in the current panel of allergens for molecule-based diagnosis will facilitate the evaluation of the clinical relevance of nsLTP sensitization in celery allergy and help clinicians in the management of food allergic patients.
Reshaping the Bet V 1 Fold Modulates T(H) Polarization
Several alternative mechanisms have been proposed to explain why some proteins are able to induce a T(H)2-biased and IgE-mediated immune response. These include specific interactions with receptors of the innate immune system, proteolytic activities, allergen-associated carbohydrate structures, and intrinsic structural determinants.
Natalizumab Therapy Decreases Surface Expression of Both VLA-heterodimer Subunits on Peripheral Blood Mononuclear Cells
Natalizumab interferes with immune cell migration into the central nervous system via blocking the alpha-4 subunit of very-late activation antigen-4 (VLA-4). Occurrence of rare but serious progressive multifocal leukoencephalopathy during prolonged natalizumab therapy of multiple sclerosis (MS) calls for a more detailed understanding of potential coeffects. We longitudinally studied alpha-4 and beta-1 surface levels on blood cells from 18 MS patients by flow cytometry. Expectedly, detectability of natalizumab-blocked alpha-4 was diminished on all investigated cell subsets. In addition, we report a concurrent and significant decrease of beta-1 surface levels on T-cells, B-cells, natural killer cells, and natural killer T cells, but not on monocytes. Uncovering secondary effects of natalizumab is mandatory to increase safety in MS therapy.
Molecular Characterization of Api G 2, a Novel Allergenic Member of the Lipid-transfer Protein 1 Family from Celery Stalks
Celery represents a relevant cross-reactive food allergen source in the adult population. As the currently known allergens are not typical elicitors of severe symptoms, we aimed to identify and characterize a non-specific lipid transfer protein (nsLTP).
Current Therapies in Ischemic Stroke. Part A. Recent Developments in Acute Stroke Treatment and in Stroke Prevention
Stroke is the third leading cause of death with an increasing prevalence. In previous years many important achievements and new therapeutic strategies have been established. This article provides an overview on recent developments and is an update to the article of Green et al. that was published in 2004. As this article is a comprehensive review we divided it in two parts. In this Part A of our review, recent developments in acute stroke treatment and in stroke prevention are described. In Part B we will reflect on neuroprotection.
Recent Developments in Approved and Oral Multiple Sclerosis Treatment and an Update on Future Treatment Options
Multiple sclerosis (MS) is the most common chronic neurological disease in young adults in the western world. There was no specific treatment available for this serious disorder until the introduction of the immunomodulatory drug interferon-β in the mid-1990s. Since then, the number of agents and treatment strategies for MS has increased rapidly. Deeper knowledge on the heterogeneous nature of MS cleared the way for several more specific, more effective and more comfortable therapies. Here, because of the exciting recent developments concerning oral treatment forms for MS, we summarize the current state of approved and future therapy options. In particular, we highlight oral treatment options in MS.
