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Ball A, Khatri K, Glesner J, Vailes LD, Wünschmann S, Gabel SA, Mueller GA, Zhang J, Peebles RS, Chapman MD, Smith SA, Chruszcz M, Pomés A. Structural analysis of human IgE monoclonal antibody epitopes on dust mite allergen Der p 2. J Allergy Clin Immunol 2024; 154:447-457. [PMID: 38697404 PMCID: PMC11409219 DOI: 10.1016/j.jaci.2024.04.017] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2024] [Revised: 03/19/2024] [Accepted: 04/09/2024] [Indexed: 05/05/2024]
Abstract
BACKGROUND Human IgE (hIgE) mAbs against major mite allergen Der p 2 developed using human hybridoma technology were used for IgE epitope mapping and analysis of epitopes associated with the hIgE repertoire. OBJECTIVE We sought to elucidate the new hIgE mAb 4C8 epitope on Der p 2 and compare it to the hIgE mAb 2F10 epitope in the context of the allergenic structure of Der p 2. METHODS X-ray crystallography was used to determine the epitope of anti-Der p 2 hIgE mAb 4C8. Epitope mutants created by targeted mutagenesis were analyzed by immunoassays and in vivo using a human high-affinity IgE receptor (FcεRIα)-transgenic mouse model of passive systemic anaphylaxis. RESULTS The structure of recombinant Der p 2 with hIgE mAb 4C8 Fab was determined at 3.05 Å. The newly identified epitope region does not overlap with the hIgE mAb 2F10 epitope or the region recognized by 3 overlapping hIgE mAbs (1B8, 5D10, and 2G1). Compared with wild-type Der p 2, single or double 4C8 and 2F10 epitope mutants bound less IgE antibodies from allergic patients by as much as 93%. Human FcεRIα-transgenic mice sensitized by hIgE mAbs, which were susceptible to anaphylaxis when challenged with wild-type Der p 2, could no longer cross-link FcεRI to induce anaphylaxis when challenged with the epitope mutants. CONCLUSIONS These data establish the structural basis of allergenicity of 2 hIgE mAb nonoverlapping epitopes on Der p 2, which appear to make important contributions to the hIgE repertoire against Der p 2 and provide molecular targets for future design of allergy therapeutics.
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Affiliation(s)
| | - Kriti Khatri
- Michigan State University, East Lansing, Mich; University of South Carolina, Columbia, SC
| | | | | | | | - Scott A Gabel
- National Institute of Environmental Health Sciences, Research Triangle Park, NC
| | - Geoffrey A Mueller
- National Institute of Environmental Health Sciences, Research Triangle Park, NC
| | - Jian Zhang
- Vanderbilt University Medical Center, Nashville, Tenn
| | | | | | - Scott A Smith
- Vanderbilt University Medical Center, Nashville, Tenn
| | - Maksymilian Chruszcz
- Michigan State University, East Lansing, Mich; University of South Carolina, Columbia, SC.
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2
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Hesse L, Oude Elberink J, van Oosterhout AJ, Nawijn MC. Allergen immunotherapy for allergic airway diseases: Use lessons from the past to design a brighter future. Pharmacol Ther 2022; 237:108115. [DOI: 10.1016/j.pharmthera.2022.108115] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2021] [Revised: 12/23/2021] [Accepted: 01/11/2022] [Indexed: 10/19/2022]
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Popescu FD, Vieru M. Precision medicine allergy immunoassay methods for assessing immunoglobulin E sensitization to aeroallergen molecules. World J Methodol 2018; 8:17-36. [PMID: 30519536 PMCID: PMC6275558 DOI: 10.5662/wjm.v8.i3.17] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/08/2018] [Revised: 08/17/2018] [Accepted: 10/09/2018] [Indexed: 02/06/2023] Open
Abstract
Molecular-based allergy diagnosis for the in vitro assessment of a patient immunoglobulin E (IgE) sensitization profile at the molecular level uses allergen molecules (also referred to as allergen components), which may be well-defined, highly purified, natural allergen components or recombinant allergens. Modern immunoassay methods used for the detection of specific IgE against aeroallergen components are either singleplex (such as the fluorescence enzyme immunoassay with capsulated cellulose polymer solid-phase coupled allergens, the enzyme-enhanced chemiluminescence immunoassay and the reversed enzyme allergosorbent test, with liquid-phase allergens), multiparameter (such as the line blot immunoassay for defined partial allergen diagnostics with allergen components coating membrane strips) or multiplex (such as the microarray-based immunoassay on immuno solid-phase allergen chip, and the two new multiplex nanotechnology-based immunoassays: the patient-friendly allergen nano-bead array, and the macroarray nanotechnology-based immunoassay used as a molecular allergy explorer). The precision medicine diagnostic work-up may be organized as an integrated “U-shape” approach, with a “top-down” approach (from symptoms to molecules) and a “bottom-up” approach (from molecules to clinical implications), as needed in selected patients. The comprehensive and accurate IgE sensitization molecular profiling, with identification of the relevant allergens, is indicated within the framework of a detailed patient’s clinical history to distinguish genuine IgE sensitization from sensitization due to cross-reactivity (especially in polysensitized patients), to assess unclear symptoms and unsatisfactory response to treatment, to reveal unexpected sensitizations, and to improve assessment of severity and risk aspects in some patients. Practical approaches, such as anamnesis molecular thinking, laboratory molecular thinking and postmolecular anamnesis, are sometimes applied. The component-resolved diagnosis of the specific IgE repertoire has a key impact on optimal decisions making for prophylactic and specific immunotherapeutic strategies tailored for the individual patient.
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Affiliation(s)
- Florin-Dan Popescu
- Department of Allergology, “Carol Davila” University of Medicine and Pharmacy, Bucharest 022441, Romania
- Department of Allergology and Clinical Immunology, “Nicolae Malaxa” Clinical Hospital, Bucharest 022441, Romania
| | - Mariana Vieru
- Department of Allergology, “Carol Davila” University of Medicine and Pharmacy, Bucharest 022441, Romania
- Department of Allergology and Clinical Immunology, “Nicolae Malaxa” Clinical Hospital, Bucharest 022441, Romania
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Su Y, Romeu-Bonilla E, Heiland T. Next generation immunotherapy for tree pollen allergies. Hum Vaccin Immunother 2018; 13:2402-2415. [PMID: 28853984 DOI: 10.1080/21645515.2017.1367882] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022] Open
Abstract
Tree pollen induced allergies are one of the major medical and public health burdens in the industrialized world. Allergen-Specific Immunotherapy (AIT) through subcutaneous injection or sublingual delivery is the only approved therapy with curative potential to pollen induced allergies. AIT often is associated with severe side effects and requires long-term treatment. Safer, more effective and convenient allergen specific immunotherapies remain an unmet need. In this review article, we discuss the current progress in applying protein and peptide-based approaches and DNA vaccines to the clinical challenges posed by tree pollen allergies through the lens of preclinical animal models and clinical trials, with an emphasis on the birch and Japanese red cedar pollen induced allergies.
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Affiliation(s)
- Yan Su
- a Department of R&D , Immunomic Therapeutics, Inc. (ITI) , Rockville , MD , USA
| | | | - Teri Heiland
- a Department of R&D , Immunomic Therapeutics, Inc. (ITI) , Rockville , MD , USA
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Tscheppe A, Breiteneder H. Recombinant Allergens in Structural Biology, Diagnosis, and Immunotherapy. Int Arch Allergy Immunol 2017; 172:187-202. [PMID: 28467993 DOI: 10.1159/000464104] [Citation(s) in RCA: 33] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/15/2023] Open
Abstract
The years 1988-1995 witnessed the beginning of allergen cloning and the generation of recombinant allergens, which opened up new avenues for the diagnosis and research of human allergic diseases. Most crystal and solution structures of allergens have been obtained using recombinant allergens. Structural information on allergens allows insights into their evolutionary biology, illustrates clinically observed cross-reactivities, and makes the design of hypoallergenic derivatives for allergy vaccines possible. Recombinant allergens are widely used in molecule-based allergy diagnosis such as protein microarrays or suspension arrays. Recombinant technologies have been used to produce well-characterized, noncontaminated vaccine components with known biological activities including a variety of allergen derivatives with reduced IgE reactivity. Such recombinant hypoallergens as well as wild-type recombinant allergens have been used successfully in several immunotherapy trials for more than a decade to treat birch and grass pollen allergy. As a more recent application, the development of antibody repertoires directed against conformational epitopes during immunotherapy has been monitored by recombinant allergen chimeras. Although much progress has been made, the number and quality of recombinant allergens will undoubtedly increase and keep improving our knowledge in basic scientific investigations, diagnosis, and therapy of human allergic diseases.
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Affiliation(s)
- Angelika Tscheppe
- Department of Pathophysiology and Allergy Research, Medical University of Vienna, Vienna, Austria
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Moingeon P, Floch VBL, Airouche S, Baron-Bodo V, Nony E, Mascarell L. Allergen immunotherapy for birch pollen-allergic patients: recent advances. Immunotherapy 2016; 8:555-67. [DOI: 10.2217/imt-2015-0027] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022] Open
Abstract
As of today, allergen immunotherapy is performed with aqueous natural allergen extracts. Recombinant allergen vaccines are not yet commercially available, although they could provide patients with well-defined and highly consistent drug substances. As Bet v 1 is the major allergen involved in birch pollen allergy, with more than 95% of patients sensitized to this allergen, pharmaceutical-grade recombinant Bet v 1-based vaccines were produced and clinically tested. Herein, we compare the clinical results and modes of action of treatments based on either a birch pollen extract or recombinant Bet v 1 expressed as hypoallergenic or natural-like molecules. We also discuss the future of allergen immunotherapy with improved drugs intended for birch pollen-allergic patients suffering from rhinoconjunctivitis.
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Affiliation(s)
- Philippe Moingeon
- Stallergenes Greer, Research Department, 6 rue Alexis de Tocqueville, 92183 Antony Cedex, France
| | | | - Sabi Airouche
- Stallergenes Greer, Research Department, 6 rue Alexis de Tocqueville, 92183 Antony Cedex, France
| | - Véronique Baron-Bodo
- Stallergenes Greer, Research Department, 6 rue Alexis de Tocqueville, 92183 Antony Cedex, France
| | - Emmanuel Nony
- Stallergenes Greer, Research Department, 6 rue Alexis de Tocqueville, 92183 Antony Cedex, France
| | - Laurent Mascarell
- Stallergenes Greer, Research Department, 6 rue Alexis de Tocqueville, 92183 Antony Cedex, France
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Abstract
Presently, allergy diagnosis and therapy procedures are undergoing a transition phase in which allergen extracts are being step-by-step replaced by molecule-based products. The new developments will allow clinicians to obtain detailed information on sensitization patterns, more accurate interpretation of allergic symptoms, and thus improved patients' management. In this respect, recombinant technology has been applied to develop this new generation of molecule-based allergy products. The use of recombinant allergens allows full validation of identity, quantity, homogeneity, structure, aggregation, solubility, stability, IgE-binding and the biologic potency of the products. In contrast, such parameters are extremely difficult to assay and standardize for extract-based products. In addition to the possibility of bulk production of wild type molecules for diagnostic purposes, recombinant technology opened the possibility of developing safer and more efficacious products for allergy therapy. A number of molecule-based hypoallergenic preparations have already been successfully evaluated in clinical trials, bringing forward the next generation of allergy vaccines. In this contribution, we review the latest developments in allergen characterization, molecule-based allergy diagnosis, and the application of recombinant allergens in therapeutic setups. A comprehensive overview of clinical trials using recombinant allergens as well as synthetic peptides is presented.
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Affiliation(s)
- Fatima Ferreira
- Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
| | - Martin Wolf
- Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria
| | - Michael Wallner
- Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria
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Niederberger V, Eckl-Dorna J, Pauli G. Recombinant allergen-based provocation testing. Methods 2014; 66:96-105. [PMID: 23920475 PMCID: PMC3988965 DOI: 10.1016/j.ymeth.2013.07.037] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/14/2013] [Revised: 07/08/2013] [Accepted: 07/15/2013] [Indexed: 01/02/2023] Open
Abstract
Over the last 25 years, recombinant allergens from all important allergen sources have been cloned and are now available as recombinant proteins. These molecules can be produced in practically unlimited amounts without biological or batch-to-batch variability. It has been shown in provocation tests that recombinant allergens have similar clinical effects as their natural counterparts. With the help of these tools it is possible to reveal the precise reactivity profiles of patients and to uncover and differentiate cross-reactivity from genuine sensitization to an allergen source. Although it has been shown some time ago that it would be possible to replace crude allergen extracts with recombinant allergens for skin prick testing, and even though the use of allergen components can improve routine diagnosis, these tools are still not available for clinical routine applications. The use of provocation tests is a crucial step in the development of new, hypoallergenic vaccines for therapy of allergic disease. Here we describe important provocation methods (skin prick test, intradermal test, atopy patch test, nasal provocation, colonoscopic provocation test) and give an overview of the clinical provocation studies which have been performed with recombinant allergens so far.
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Affiliation(s)
| | - Julia Eckl-Dorna
- Dept. of Otorhinolaryngology, Medical University of Vienna, Austria
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Spertini F, Reymond C, Leimgruber A. Allergen-specific immunotherapy of allergy and asthma: current and future trends. Expert Rev Respir Med 2012; 3:37-51. [PMID: 20477281 DOI: 10.1586/17476348.3.1.37] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Allergen-specific immunotherapy is the only immunomodulatory and etiological therapy of allergy and asthma. Conventional specific immunotherapy (SIT) with whole-allergen extract is antigen specific, effective on multiple organs, efficient on asthma in defined conditions, provides long-lasting protection and is cost effective. Moreover, SIT is able to prevent the course of rhinitis to asthma. SIT has its drawbacks: the long duration of treatment, the unsatisfactory standardization of allergen extracts and a questionable safety level. Novel approaches are aimed at drastically reducing adverse anaphylactic events, shortening the duration of therapy and improving its efficacy. Novel promising approaches have based their formulation on a limited set of recombinant allergens or chimeric molecules as well as on hypoallergenic allergen fragments or peptides. The simultaneous use of adjuvants with immunomodulatory properties may contribute to improve both the safety and efficacy of allergen-SIT of allergy and asthma.
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Affiliation(s)
- François Spertini
- Division of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, Rue du Bugnon 46, 1011 Lausanne, Switzerland.
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Jutel M, Solarewicz-Madejek K, Smolinska S. Recombinant allergens: the present and the future. Hum Vaccin Immunother 2012; 8:1534-43. [PMID: 23095874 DOI: 10.4161/hv.22064] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023] Open
Abstract
Allergen specific immunotherapy (SIT) is the only known causative treatment of allergic diseases. Recombinant allergen-based vaccination strategies arose from a strong need to both to improve safety and enhance efficacy of SIT. In addition, new vaccines can be effective in allergies including food allergy or atopic dermatitis, which poorly respond to the current treatment with allergen extracts. A number of successful clinical studies with both wild-type and hypoallergenic derivatives of recombinant allergens vaccines have been reported for the last decade. They showed high efficacy and safety profile as well as very strong modulation of T and B cell responses to specific allergens.
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Affiliation(s)
- Marek Jutel
- Department of Clinical Immunology, Wroclaw Medical University, Wroclaw, Poland.
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12
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Allergen-specific immunotherapy with recombinant allergens. Curr Top Microbiol Immunol 2012; 352:43-54. [PMID: 21404096 DOI: 10.1007/82_2011_125] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
Subcutaneous immunotherapy is a well-documented treatment of allergic rhinitis and asthma. The major limitation is the risk of anaphylactic side effects. The documentation of clinical efficacy is based on crude allergenic extracts sometimes containing varying amounts of individual allergens including allergens to which the patient may not be sensitized. The introduction of recombinant allergens offer a possibility to use well-defined molecules with consistent pharmaceutical quality defined in mass units. The proof-of-concept of the clinical efficacy of recombinant allergens is based on two studies published as full articles. One study applied a mixture of five Phleum pratense major allergens in a maximum dose of 40 μg protein. The clinical efficacy showed a significant efficacy with about 40% reduction in disease severity. The second study compared a commercial birch extract with both recombinant Bet v 1 and purified Bet v 1 in dosages of 15 μg allergen. The clinical effect was around 60% additional efficacy. Systemic side effects occurred more frequently with grass allergens. A third study used hypoallergenic fragments and a trimer of Bet v 1. The study did not show efficacy and a rather high frequency of systemic side effects. The advantages of using recombinant allergens for immunotherapy are obvious but more large-scale clinical studies are needed before the overall value in terms of efficacy and safety can be determined.
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Linhart B, Valenta R. Mechanisms underlying allergy vaccination with recombinant hypoallergenic allergen derivatives. Vaccine 2011; 30:4328-35. [PMID: 22100888 DOI: 10.1016/j.vaccine.2011.11.011] [Citation(s) in RCA: 52] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2011] [Revised: 10/24/2011] [Accepted: 11/03/2011] [Indexed: 10/15/2022]
Abstract
Hundred years ago therapeutic vaccination with allergen-containing extracts has been introduced as a clinically effective, disease-modifying, allergen-specific and long-lasting form of therapy for allergy, a hypersensitivity disease affecting more than 25% of the population. Today, the structures of most of the disease-causing allergens have been elucidated and recombinant hypoallergenic allergen derivatives with reduced allergenic activity have been engineered to reduce side effects during allergen-specific immunotherapy (SIT). These recombinant hypoallergens have been characterized in vitro, in experimental animal models and in clinical trials in allergic patients. This review provides a summary of the molecular, immunological and preclinical evaluation criteria applied for this new generation of allergy vaccines. Furthermore, we summarize the mechanisms underlying SIT with recombinant hypoallergens which are thought to be responsible for their therapeutic effect.
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Affiliation(s)
- Birgit Linhart
- Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
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Valenta R, Linhart B, Swoboda I, Niederberger V. Recombinant allergens for allergen-specific immunotherapy: 10 years anniversary of immunotherapy with recombinant allergens. Allergy 2011; 66:775-83. [PMID: 21352238 DOI: 10.1111/j.1398-9995.2011.02565.x] [Citation(s) in RCA: 109] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
Abstract
The broad applicability of allergen-specific immunotherapy for the treatment and eventually prevention of IgE-mediated allergy is limited by the poor quality and allergenic activity of natural allergen extracts that are used for the production of current allergy vaccines. Today, the genetic code of the most important allergens has been deciphered; recombinant allergens equalling their natural counterparts have been produced for diagnosis and immunotherapy, and a large panel of genetically modified allergens with reduced allergenic activity has been characterized to improve safety of immunotherapy and explore allergen-specific prevention strategies. Successful immunotherapy studies have been performed with recombinant allergens and hypoallergenic allergen derivatives and will lead to the registration of the first recombinant allergen-based vaccines in the near future. There is no doubt that recombinant allergen-based vaccination strategies will be generally applicable to most allergen sources, including respiratory, food and venom allergens and allow to produce safe allergy vaccines for the treatment of the most common forms of IgE-mediated allergies.
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Affiliation(s)
- Rudolf Valenta
- Christian Doppler Laboratory for Allergy Research, Vienna, Austria.
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The current state of recombinant allergens for immunotherapy. Curr Opin Allergy Clin Immunol 2011; 10:575-81. [PMID: 20859201 DOI: 10.1097/aci.0b013e32833fd6c5] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
PURPOSE OF REVIEW Subcutaneous immunotherapy is a well documented treatment of allergic rhinitis and asthma. The majority of the disadvantages of the treatment are related to the poor quality of the natural allergen extracts which can contain varying amounts of individual allergens including allergens to which the patient may not be sensitized. Recombinant allergens offer a possibility to use well defined molecules with consistent pharmaceutical quality defined in mass units. The proof of concept of the clinical efficacy of recombinant allergens is based on two studies published as full articles. RECENT FINDINGS One study applied a mixture of five Phleum pratense major allergens in a maximum dose of 40mcg protein. The clinical efficacy showed a significant efficacy with 40% reduction in disease severity. The second study compared a commercial birch extract with both recombinant Bet v 1 and purified Bet v 1 in dosages of 15mcg allergen. The clinical effect was 60% additional efficacy. Systemic side effects occurred more frequently with grass allergens. A third study used hypoallergenic fragments and a trimer of Bet v 1. The study did not show efficacy and a rather high frequency of systemic side effects. SUMMARY The advantages of using recombinant allergens for immunotherapy are obvious but more studies on a large scale are needed before the overall value in terms of efficacy and safety can be assessed. Clinical trials are also necessary for new combined vaccines based on recombinant allergens that in experimental studies have shown greatly enhanced immunogenicity and low allergen-specific reactivity.
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Recombinant allergens for specific immunotherapy. J Allergy Clin Immunol 2011; 127:865-72. [DOI: 10.1016/j.jaci.2011.01.047] [Citation(s) in RCA: 76] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2010] [Revised: 12/16/2010] [Accepted: 01/21/2011] [Indexed: 12/11/2022]
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Clinical practice: Allergen-specific immunotherapy in children: facts and FAQs. Eur J Pediatr 2011; 170:137-48. [PMID: 21153032 DOI: 10.1007/s00431-010-1348-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/13/2010] [Accepted: 11/04/2010] [Indexed: 10/18/2022]
Abstract
Allergen-specific immunotherapy (SIT) in its various application forms represents the main treatment approach of IgE-mediated allergic diseases in adults and children. Despite this clear recommendation, many particularities of products, patient characteristics, and product availability in different countries hamper the use of allergen-specific immunotherapy in particular in children. The frequently asked questions by parents, patients, and physicians are the backbone of this review. Thus, the potentials and limitations of allergen-specific immunotherapy in children and adolescents will be highlighted. IgE-mediated allergic diseases are affecting about 20% of the population. They manifest commonly early in life, and hence, the use of SIT should be considered also early in the course of the disease.
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Vrtala S, Fohr M, Campana R, Baumgartner C, Valent P, Valenta R. Genetic engineering of trimers of hypoallergenic fragments of the major birch pollen allergen, Bet v 1, for allergy vaccination. Vaccine 2011; 29:2140-8. [PMID: 21215346 DOI: 10.1016/j.vaccine.2010.12.080] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2010] [Revised: 12/13/2010] [Accepted: 12/20/2010] [Indexed: 10/18/2022]
Abstract
An immunotherapy trial performed in allergic patients with hypoallergenic recombinant fragments, comprising aa 1-74 and 75-160 of the major birch pollen allergen, Bet v 1, has indicated that the induction of allergen-specific IgG responses may be an important mechanism of this treatment. To investigate whether the immunogenicity of the rBet v 1 fragments can be increased, recombinant trimers of the fragments were produced. For this purpose, DNA trimers of rBet v 1 aa 1-74 as well as of rBet v 1 aa 75-160 were subcloned into expression plasmid pET 17b, expressed in Escherichia coli and purified. The fragments as well as the fragment trimers showed a reduced IgE-binding capacity and allergenic activity compared to rBet v 1 wildtype when tested in allergic patients. Both rBet v 1 aa 75-160 monomer and trimer induced high titers of allergen-specific IgG1 Abs in mice. Interestingly, rBet v 1 aa 1-74 trimer induced a much higher IgG(1) response to rBet v 1 than rBet v 1 aa 1-74 monomer. Consequently, IgG Abs induced with the rBet v 1 aa 1-74 trimer inhibited birch pollen allergic patients' IgE-binding 10-fold more efficiently than IgG Abs induced with the monomer. Our data show that the immunogenicity of allergy vaccines can be increased by oligomerization.
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Affiliation(s)
- Susanne Vrtala
- Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria
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Cromwell O, Niederberger V, Horak F, Fiebig H. Clinical Experience with Recombinant Molecules for Allergy Vaccination. Curr Top Microbiol Immunol 2011; 352:27-42. [DOI: 10.1007/82_2011_129] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
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Campana R, Vrtala S, Maderegger B, Jertschin P, Stegfellner G, Swoboda I, Focke-Tejkl M, Blatt K, Gieras A, Zafred D, Neubauer A, Valent P, Keller W, Spitzauer S, Valenta R. Hypoallergenic derivatives of the major birch pollen allergen Bet v 1 obtained by rational sequence reassembly. J Allergy Clin Immunol 2010; 126:1024-31, 1031.e1-8. [PMID: 20638112 DOI: 10.1016/j.jaci.2010.05.023] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2009] [Revised: 05/10/2010] [Accepted: 05/18/2010] [Indexed: 12/23/2022]
Abstract
BACKGROUND At least 100 million patients suffer from birch pollen allergy. OBJECTIVE Rational design of recombinant derivatives of the major birch pollen allergen, Bet v 1, characterized by reduced IgE reactivity, preservation of sequences relevant for the induction of allergen-specific blocking IgG, and maintenance of T-cell epitopes for immunotherapy of birch pollen allergy. METHODS Three recombinant mosaic proteins derived from Bet v 1 were generated by reassembly of codon-optimized genes coding for Bet v 1 fragments containing the elements for the induction of allergen-specific blocking IgG antibodies and the major T-cell epitopes. The proteins were expressed in Escherichia coli as recombinant mosaic molecules and compared with the Bet v 1 wild-type protein by chemical and structural methods, regarding IgE-binding and IgG-binding capacity, in basophil activation assays and tested for the in vivo induction of IgG responses. RESULTS Three recombinant Bet v 1 (rBet v 1) mosaic proteins with strongly reduced IgE reactivity and allergenic activity were expressed and purified. Immunization with the recombinant hypoallergens induced IgG antibodies that inhibited IgE reactivity of patients with allergy to Bet v 1 comparable to those induced with the rBet v 1 wild-type allergen. CONCLUSION We report the generation and preclinical characterization of 3 hypoallergenic rBet v 1 derivatives with suitable properties for immunotherapy of birch pollen allergy.
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Affiliation(s)
- Raffaela Campana
- Division of Immunopathology, Department of Pathophysiology, Center of Pathophysiology, Infectiology and Immunology, Vienna General Hospital (AKH), Medical University of Vienna, Vienna, Austria
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Assarehzadegan MA, Sankian M, Jabbari F, Tehrani M, Varasteh A. Expression of the recombinant major allergen of Salsola kali pollen (Sal k 1) and comparison with its low-immunoglobulin E-binding mutant. Allergol Int 2010; 59:213-22. [PMID: 20414052 DOI: 10.2332/allergolint.09-oa-0155] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2009] [Accepted: 01/14/2010] [Indexed: 11/20/2022] Open
Abstract
BACKGROUND The inhalation of Salsola kali pollen is an important cause of pollinosis during summer and early fall throughout desert and semi-desert areas. Sal k 1 has been previously reported as a major allergen of S. kali pollen. In this study, we produced the recombinant Sal k 1 and also its low IgE-binding mutant form. We further compared the IgE binding ability of these two recombinant molecules. METHODS The recombinant Sal k 1 and its low IgE-binding variant, obtained by three amino acid exchanges (R(142)-->S, P(143)-->A, D(144)-->V), were cloned and expressed in E. coli, as proteins fused with thioredoxin and His-tags, and then purified by Ni2+ affinity chromatography. The IgE-binding capacity of the wild-type and mutated rSal k 1 was compared using immunoblotting, ELISA and inhibition assays by ten sera from S. kali allergic patients. Moreover, in vivo IgE-reactivity was investigated by the skin prick test. RESULTS Both the recombinant and the mutated form of Sal k 1 were expressed in E. coli at a relatively high amount and soluble form. All sera recognized rSal k 1 via immunoassay analysis. In addition, inhibition assays demonstrated that the purified rSal k 1 was similar to its counterpart in the crude extract. The mutated rSal k 1 exhibited a reduced IgE-binding capacity against wild-type rSal k 1. CONCLUSIONS This study demonstrates that purified rSal k 1 is comprised of IgE-epitopes similar to that of its natural counterpart and that the mutated variant showed a reduced IgE-binding capacity based on in vitro assays and in vivo provocation testing.
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MESH Headings
- Adult
- Antigens, Plant/genetics
- Antigens, Plant/immunology
- Antigens, Plant/metabolism
- Chromatography, Affinity
- Cloning, Molecular
- Female
- Humans
- Immunoglobulin E/metabolism
- Male
- Mutagenesis, Site-Directed
- Mutation/genetics
- Pollen
- Protein Binding
- Recombinant Fusion Proteins/genetics
- Recombinant Fusion Proteins/immunology
- Recombinant Fusion Proteins/metabolism
- Rhinitis, Allergic, Seasonal/genetics
- Rhinitis, Allergic, Seasonal/immunology
- Rhinitis, Allergic, Seasonal/metabolism
- Salsola
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22
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Valenta R, Ferreira F, Focke-Tejkl M, Linhart B, Niederberger V, Swoboda I, Vrtala S. From allergen genes to allergy vaccines. Annu Rev Immunol 2010; 28:211-41. [PMID: 20192803 DOI: 10.1146/annurev-immunol-030409-101218] [Citation(s) in RCA: 158] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
IgE-mediated allergy is a hypersensitivity disease affecting more than 25% of the population. The structures of the most common allergens have been revealed through molecular cloning technology in the past two decades. On the basis of this knowledge of the sequences and three-dimensional structures of culprit allergens, investigators can now analyze the immune recognition of allergens and the mechanisms of allergic inflammation in allergic patients. Allergy vaccines have been constructed that are able to selectively target the aberrant immune responses in allergic patients via different pathways of the immune system. Here we review various types of allergy vaccines that have been developed based on allergen structures, results from their clinical application in allergic patients, and future strategies for allergen-specific immunotherapy and allergy prophylaxis.
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Abstract
Type I allergy represents an important health problem that affects more than 25% of the population in industrialized countries. Specific immunotherapy is one of the few causative treatment approaches for type I allergy and is currently performed with crude allergen extracts, which consist of a mixture of allergenic and nonallergenic components, are difficult to standardize and cannot be applied according to the patient's reactivity profile. With the introduction of molecular biological techniques into allergy research, a large panel of individual recombinant allergens has become available. Recombinant allergens can be used for improved diagnosis of allergy to determine the patient's sensitization profile, which is a prerequisite to select the allergens for patient-tailored immunotherapy. They allow the elucidation of the properties of allergens and of the mechanisms of allergy as well as of the mechanisms of immunotherapy. Moreover, recombinant allergens allow the development of hypoallergenic allergen derivatives with reduced allergenic activity and retained immunogenicity. First immunotherapy trials with hypoallergenic allergen derivatives have shown that this treatment might improve immunotherapy in the near future. This review summarizes the results, which were obtained with recombinant allergens and hypoallergenic allergen derivatives. The experiences from the in vitro and in vivo evaluation of the hypoallergenic derivatives and from clinical studies as well as the contribution of hypoallergenic derivatives to develop new treatment strategies and possibly prophylactic vaccination strategies are discussed.
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Affiliation(s)
- S Vrtala
- Division of Immunopathology, Department for Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria
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24
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Pree I, Reisinger J, Focke M, Vrtala S, Pauli G, van Hage M, Cromwell O, Gadermaier E, Egger C, Reider N, Horak F, Valenta R, Niederberger V. Analysis of Epitope-Specific Immune Responses Induced by Vaccination with Structurally Folded and Unfolded Recombinant Bet v 1 Allergen Derivatives in Man. THE JOURNAL OF IMMUNOLOGY 2007; 179:5309-16. [PMID: 17911617 DOI: 10.4049/jimmunol.179.8.5309] [Citation(s) in RCA: 41] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
Previously, we have constructed recombinant derivatives of the major birch pollen allergen, Bet v 1, with a more than 100-fold reduced ability to induce IgE-mediated allergic reactions. These derivatives differed from each other because the two recombinant Bet v 1 fragments represented unfolded molecules whereas the recombinant trimer resembled most of the structural fold of the Bet v 1 allergen. In this study, we analyzed the Ab (IgE, IgG subclass, IgA, IgM) response to Bet v 1, recombinant and synthetic Bet v 1-derived peptides in birch pollen allergic patients who had been vaccinated with the derivatives or adjuvant alone. Furthermore, we studied the induction of IgE-mediated skin responses in these patients using Bet v 1 and Bet v 1 fragments. Both types of vaccines induced a comparable IgG1 and IgG4 response against new sequential epitopes which overlap with the conformational IgE epitopes of Bet v 1. This response was 4- to 5-fold higher than that induced by immunotherapy with birch pollen extract. Trimer more than fragments induced also IgE responses against new epitopes and a transient increase in skin sensitivity to the fragments at the beginning of therapy. However, skin reactions to Bet v 1 tended to decrease one year after treatment in both actively treated groups. We demonstrate that vaccination with folded and unfolded recombinant allergen derivatives induces IgG Abs against new epitopes. These data may be important for the development of therapeutic as well as prophylactic vaccines based on recombinant allergens.
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Affiliation(s)
- Ines Pree
- Department of Otolaryngology, Vienna General Hospital, Medical University of Vienna, Vienna, Austria
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25
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Valenta R, Niederberger V. Recombinant allergens for immunotherapy. J Allergy Clin Immunol 2007; 119:826-30. [PMID: 17335886 DOI: 10.1016/j.jaci.2007.01.025] [Citation(s) in RCA: 127] [Impact Index Per Article: 7.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2007] [Accepted: 01/12/2007] [Indexed: 11/19/2022]
Abstract
Recombinant allergens can be produced as defined molecules in consistent quality and unlimited amounts according to the corresponding DNA template. Furthermore, they can be modified to reduce their allergenic activity and to foster certain advantageous immunologic properties. Recombinant allergens equaling the natural allergens are available for diagnostic and therapeutic purposes, and modified versions have been developed with the aim to to reduce IgE-mediated side effects during immunotherapy. First injection immunotherapy trials conducted with recombinant vaccines for birch pollen and grass pollen allergy show that recombinant allergen-based immunotherapy has vaccination characteristics and is clinically effective. The obtained results hold promise that recombinant allergen-based immunotherapy will improve current immunotherapy practice and may open possibilities for new treatment strategies and possibly even for prophylactic vaccination.
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Affiliation(s)
- Rudolf Valenta
- Christian Doppler Laboratory for Allergy Research, Division of Immunopathology, Department of Pathophysiology, Center of Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
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26
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Breiteneder H, Mills C. Structural bioinformatic approaches to understand cross-reactivity. Mol Nutr Food Res 2006; 50:628-32. [PMID: 16764018 DOI: 10.1002/mnfr.200500274] [Citation(s) in RCA: 35] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Cross-reactivity of allergens results from the presence of antibody-accessible conserved surface structures. These can best be studied when allergens have been structurally defined by X-ray crystallography or another structure determination method. When this is not the case, mimotope technology provides a useful alternative for elucidating antibody-binding sites on allergens. Structural bioinformatic approaches have been used to study the cross-reactivity of inhalant allergens with labile food allergens (Bet v 1 family) as well as the cross-reactivity between stable food allergens such as members of the nonspecific lipid transfer protein family. It was found that the degree of similarity of the structures correlated with the observed IgE cross-reactivities. However, IgE cross-reactivity between structurally unrelated allergens has not been demonstrated to date.
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Affiliation(s)
- Heimo Breiteneder
- Department of Pathophysiology, Medical University of Vienna, Vienna, Austria.
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Cromwell O, Fiebig H, Suck R, Kahlert H, Nandy A, Kettner J, Narkus A. Strategies for recombinant allergen vaccines and fruitful results from first clinical studies. Immunol Allergy Clin North Am 2006; 26:261-81, vii. [PMID: 16701144 DOI: 10.1016/j.iac.2006.02.001] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
Recombinant DNA technology has delivered the prospect of a new generation of preparations for allergen-specific immunotherapy. The first clinical studies with recombinant allergens have yielded encouraging results, suggesting that there is a good chance that such preparations will become available for use in the routine management of allergic disease.
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Affiliation(s)
- Oliver Cromwell
- Research and Development, Allergopharma Joachim Ganzer KG, Hermann-Koerner-Strasse 52, D-21465 Reinbek, Germany.
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Abstract
Type I allergy represents an important health problem that is currently affecting approximately 25% of the population in Western countries. Immunotherapy, the only causative treatment of Type I allergy, is currently performed with crude allergen extracts, which contain unpredictable amounts of allergenic, as well as nonallergenic, components. The application of molecular biology for allergen characterization has revealed the molecular nature of the most common allergens and allowed the production of recombinant allergens that equal natural allergens. Based on this knowledge, several different strategies to improve immunotherapy have become available. Until now, T-cell peptides, selected wild-type-like recombinant allergens and genetically modified hypoallergenic allergen derivatives have been evaluated in clinical trials in patients. Immunotherapy based on T-cell peptides has focused on allergen-specific T-cell responses, whereas genetically modified recombinant allergen molecules offer the advantage of combining T-cell and B-cell epitopes. Genetically modified recombinant birch pollen derivatives (Bet v 1-fragments, Bet v 1-trimer) have been evaluated in a double-blind, placebo-controlled, multicenter study. Vaccination with the Bet v 1-derivatives improved symptoms of birch pollen allergy, induced a healthy allergen-specific immunoglobulin G response and led to a significant reduction of seasonally induced boosts of immunoglobulin E.
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Affiliation(s)
- Verena Niederberger
- Department of Otolaryngology, Vienna General Hospital, AKH, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
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29
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Mothes N, Valenta R, Spitzauer S. Allergy testing: the role of recombinant allergens. Clin Chem Lab Med 2006; 44:125-32. [PMID: 16475895 DOI: 10.1515/cclm.2006.024] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
Currently, diagnosis of type I allergy is performed using crude allergen extracts, which allow the identification of the allergen-containing source responsible for type I allergic symptoms (e.g., allergic rhino-conjunctivitis, asthma) but not the disease-eliciting molecules. With the introduction of recombinant allergens produced by molecular biology techniques, a large panel of allergenic molecules has become available. The application of these recombinant allergens for in vitro tests has led to new forms of component-resolved diagnosis (CRD) and allows the establishment of a patient's individual reactivity profile. The increasing number of recombinant allergens characterized during the last decade has allowed the development of chip-based allergy tests for simultaneous detection of up to 5000 different allergens and epitopes. The introduction of these recombinant allergen-based tests into clinical practice improves the selection of patients for traditional specific immunotherapy and allows monitoring of the immunological efficacy of specific immunotherapy by measuring allergen-specific IgG antibodies. Besides their diagnostic application, recombinant allergens and hypoallergenic derivatives thereof have also been used as vaccines in clinical trials, and recent results have shown their usefulness for the treatment of type I allergy.
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Affiliation(s)
- Nadine Mothes
- Zentrum für Physiologie und Pathophysiologie, Institut für Pathophysiologie, Abteilung Immunpathologie, Medizinische Universität Wien, Vienna, Austria.
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30
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Winkler B, Hufnagl K, Spittler A, Ploder M, Kállay E, Vrtala S, Valenta R, Kundi M, Renz H, Wiedermann U. The role of Foxp3+ T cells in long-term efficacy of prophylactic and therapeutic mucosal tolerance induction in mice. Allergy 2006; 61:173-80. [PMID: 16409192 DOI: 10.1111/j.1398-9995.2006.01014.x] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/16/2023]
Abstract
BACKGROUND Mucosal tolerance induction is suggested as treatment strategy for allergic diseases. Using a murine model of birch pollen (BP) allergy we investigated the long-term efficacy and the underlying mechanisms of mucosal tolerance induction with two structurally different molecules in a prophylactic and in a therapeutic set-up. METHODS The three-dimensional major BP allergen Bet v 1 or a nonconformational hypoallergenic fragment thereof was intranasally applied before (prophylaxis) or after sensitization (therapy). RESULTS In the prophylactic application both the Bet v 1 allergen and the fragment prevented allergic sensitization, and this effect lasted for 1 year. In the therapeutic approach established allergic immune responses were also suppressed after treatment with either of the molecules. However, a long-lasting curative effect (6 months) was only achieved with the Bet v 1 allergen but not with the Bet v 1 fragment. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of splenocytes revealed that tolerance induction with the Bet v 1 allergen was associated with enhanced expression of transforming growth factor (TGF)-beta, interleukin (IL)-10, and Foxp3 mRNA in CD4+ T cells, whereas treatment with the fragment led to the induction of either Foxp3 (prophylaxis) or IL-10 (therapy) alone. CONCLUSION From these data we concluded (i) that the mechanisms underlying peripheral tolerance are linked to the conformation of the antigen, (ii) that mucosal tolerance is mediated by separate regulatory cell subsets, and (iii) that the long-term efficacy of immunosuppression is associated with the presence of Foxp3+ T cells.
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Affiliation(s)
- B Winkler
- Department of Specific Prophylaxis and Tropical Medicine, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria
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31
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Abstract
The application of recombinant DNA technology to allergen research has provided the sequence information and genetic material to produce new types of allergy vaccines. One general strategy has been to use the knowledge to produce synthetic peptides that represent selected T-cell or B-cell epitopes. The production of genetically engineered allergens provides an alternative strategy to construct hypoallergenic vaccines, which can provide a better and less selected representation of the epitopes. Many strategies have been used to produce such hypoallergens, and their ability to reduce allergenicity has been amply demonstrated by skin and nasal provocation tests. The retention of T cell-stimulating activity has also been demonstrated, and a consistent feature of the vaccines has been, despite the reduced immunoglobulin E (IgE)-binding reactivity, the ability to induce anti-allergen IgG antibody. The lead hypoallergens have been polypeptide fragments and trimeric constructs of the birch allergen Bet v 1. A clinical trial with these medicaments has shown the ability to modify IgE and IgG antibody production, skin test reactivity, and symptom scores. This is the first trial of a recombinant allergy vaccine, and it has set a benchmark for further studies. A new generation of hypoallergens is now being produced based on the detailed knowledge of the tertiary structures of the allergens and of the T-cell and B-cell epitopes. The modifications have been made to change the topography of the allergens while retaining a stable, folding structure. In the case of Bet v 1, tertiary structures of hypoallergens have been determined. Structurally modeled hypoallergens have been produced for pollen, venom, food, and latex allergens, with promising characteristics from preclinical studies.
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Affiliation(s)
- Wayne R Thomas
- Centre for Child Health Research, University of Western Australia, Telethon Institute for Child Health Research, PO Box 855, West Perth 6872, 100 Roberts Road, Subiaco 6008, Western Australia.
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32
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Gafvelin G, Thunberg S, Kronqvist M, Grönlund H, Grönneberg R, Troye-Blomberg M, Akdis M, Fiebig H, Purohit A, Horak F, Reisinger J, Niederberger V, Akdis CA, Cromwell O, Pauli G, Valenta R, van Hage M. Cytokine and Antibody Responses in Birch-Pollen-Allergic Patients Treated with Genetically Modified Derivatives of the Major Birch Pollen Allergen Bet v 1. Int Arch Allergy Immunol 2005; 138:59-66. [PMID: 16103688 DOI: 10.1159/000087358] [Citation(s) in RCA: 60] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/11/2005] [Accepted: 04/11/2005] [Indexed: 02/02/2023] Open
Abstract
BACKGROUND Recently, recombinant hypoallergenic derivatives of the major birch pollen allergen, Bet v 1, were used to treat birch-pollen-allergic patients in a double-blind, placebo-controlled, multi-centre immunotherapy study. The aim of this study was to evaluate the effects of vaccination with aluminium-hydroxide-adsorbed recombinant Bet v 1 derivatives versus placebo on T-cell, cytokine and antibody responses in a subgroup of patients. METHODS Blood was drawn from patients of the Swedish centre (n = 27; rBet v 1 fragments: n = 10; rBet v 1 trimer: n = 8, and placebo-aluminium hydroxide: n = 9) before the start and after completion of the treatment. PBMC were stimulated with rBet v 1 and analysed for cytokine (IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-gamma)-secreting cells by ELISpot. Bet v 1-specific antibody levels in serum (IgG(1-4), IgE and IgA) were measured by ELISA. Skin prick tests with defined Bet v 1 concentrations were performed before and 10-11 months after the beginning of the study. RESULTS Bet v 1-specific IgG levels, consisting of IgG(1), IgG(2) and IgG(4), were significantly increased after treatment with recombinant allergen derivatives. Treatment with rBet v 1 trimer led to a significant (p < 0.05) reduction of Bet v 1-reactive IL-5- and IL-13-producing cells, reflecting a reduced Th2 response. In addition, a decreased number of Bet v 1-reactive IL-4 producing (p = 0.07) and an increase of IL-12-producing (p = 0.06) cells was noted in the trimer-treated patients. In contrast to placebo, active treatment resulted in significantly reduced immediate-type skin reactions to Bet v 1 even 10-11 months after treatment. CONCLUSION Vaccination with recombinant hypoallergenic Bet v 1 derivatives induces a Bet v 1-specific IgG response and leads to reduced skin reactivity in allergic patients. A reduction of Bet v 1-specific Th2 responses was observed in trimer-treated patients, which may reflect the intrinsic property of this allergen derivative.
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Affiliation(s)
- Guro Gafvelin
- Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska Institute and University Hospital, SE-171 76 Stockholm, Sweden.
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Larché M. Peptide therapy for allergic diseases: basic mechanisms and new clinical approaches. Pharmacol Ther 2005; 108:353-61. [PMID: 16014312 DOI: 10.1016/j.pharmthera.2005.05.004] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/22/2005] [Accepted: 05/16/2005] [Indexed: 11/22/2022]
Abstract
Desensitising allergen immunotherapy has been practised for many decades. Although time consuming, this form of therapy is antigen-specific and disease-modifying, in contrast to palliative pharmacotherapy. However, the use of allergen extracts containing native allergen molecules frequently results in allergic adverse reactions to treatment. Several strategies to reduce the allergenicity of therapeutic preparations, while maintaining their therapeutic benefit, are being developed. Peptide immunotherapy is one such approach. Short synthetic peptides, comprising T cell epitopes of major allergens, were unable to crosslink allergen-specific IgE molecules on basophils in vitro. Treatment of allergic volunteers with allergen peptides resulted in reduced skin, lung and nasal sensitivity to allergen challenge and improved their subjective ability to tolerate allergen exposure. Peptides reduced pro-inflammatory cytokine secretion from peripheral blood cells, whilst increasing the immunosuppressive cytokine IL-10. Furthermore, peptide therapy was associated with the induction of a population of CD4+ T cells with a suppressive functional phenotype. Thus, peptide therapy may be suitable for the antigen-specific treatment of allergic diseases.
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Affiliation(s)
- Mark Larché
- Department of Allergy and Clinical Immunology, Imperial College London, Faculty of Medicine, National Heart and Lung Institute, Dovehouse Street, London SW3 6LY, United Kingdom.
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34
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Bischoff S, Crowe SE. Gastrointestinal food allergy: new insights into pathophysiology and clinical perspectives. Gastroenterology 2005; 128:1089-113. [PMID: 15825090 DOI: 10.1053/j.gastro.2004.08.015] [Citation(s) in RCA: 179] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/04/2023]
Abstract
Adverse reactions to food that result in gastrointestinal symptoms are common in the general population; while only a minority of such individuals will have symptoms due to immunologic reactions to foods, gastrointestinal food allergies do exist in both children and adults. These immune reactions are mediated by immunoglobulin E-dependent and -independent mechanisms involving mast cells, eosinophils, and other immune cells, but the complexity of the underlying mechanisms of pathogenesis have yet to be fully defined. Knowledge of the spectrum of adverse reactions to foods that affect the digestive system, including gastrointestinal food allergy, is essential to correctly diagnose and manage the subset of patients with immunologically mediated adverse reactions to foods. Potentially fatal reactions to food necessitate careful instruction and monitoring on the part of health care workers involved in the care of individuals at risk of anaphylaxis. New methods of diagnosis and novel strategies for treatment, including immunologic modulation and the development of hypoallergenic foods, are exciting developments in the field of food allergy.
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Affiliation(s)
- Stephan Bischoff
- Department of Gastroenterology, University Medical School of Hannover, Germany
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35
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Abstract
Allergic diseases such as asthma, rhinitis, and eczema are increasing in prevalence and affect up to 15% of populations in Westernized countries. The description of Tregs as T cells that prevent development of autoimmune disease led to considerable interest in whether these Tregs were also normally involved in prevention of sensitization to allergens and whether it might be possible to manipulate Tregs for the therapy of allergic disease. Current data suggest that Th2 responses to allergens are normally suppressed by both CD4+CD25+ Tregs and IL-10 Tregs. Furthermore, suppression by these subsets is decreased in allergic individuals. In animal models, Tregs could be induced by high- or low-dose inhaled antigen, and prior induction of such Tregs prevented subsequent development of allergen sensitization and airway inflammation in inhaled challenge models. For many years, allergen-injection immunotherapy has been used for the therapy of allergic disease, and this treatment may induce IL-10 Tregs, leading to both suppression of Th2 responses and a switch from IgE to IgG4 antibody production. Improvements in allergen immunotherapy, such as peptide therapy, and greater understanding of the biology of Tregs hold great promise for the treatment and prevention of allergic disease.
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Affiliation(s)
- Douglas S Robinson
- Department of Allergy and Clinical Immunology, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London, United Kingdom
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36
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Abstract
PURPOSE OF REVIEW This review is an update regarding the development of recombinant allergens from the laboratory bench to clinical applications. Special attention will be given to the potential improvement of allergen-specific immunotherapy through the use of recombinant allergens. RECENT FINDINGS Currently used therapeutic allergen extracts suffer from several important disadvantages and therefore may be replaced by recombinant allergens in the near future. Recent studies indicate that recombinant allergen-based diagnostic tests can be used for selection of patients for immunotherapy and to analyse the mechanisms underlying immunotherapy. Furthermore, recombinant and peptide technologies have been used for the generation of allergy vaccines with reduced allergenic activity. Applying the new technologies, the vaccines can be formulated to target either B cells or T cells, or both cell types. Very recently, encouraging results were obtained in an immunotherapy trial performed with genetically engineered allergens. SUMMARY Recombinant allergen-based diagnostic tests will improve the selection of patients for immunotherapy. The first immunotherapy trial with recombinant allergens provides information about mechanisms underlying immunotherapy and holds promise that new types of allergy vaccines based on recombinant allergens will become available soon.
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Affiliation(s)
- Verena Niederberger
- Department of Otorhinolaryngology, Centre for Physiology and Pathophysiology, Vienna General Hospital, Medical University of Vienna, A-1090 Vienna, Austria
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37
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Abstract
Allergic diseases such as asthma, rhinitis, and eczema are increasing in prevalence and affect up to 15% of populations in Westernized countries. The description of Tregs as T cells that prevent development of autoimmune disease led to considerable interest in whether these Tregs were also normally involved in prevention of sensitization to allergens and whether it might be possible to manipulate Tregs for the therapy of allergic disease. Current data suggest that Th2 responses to allergens are normally suppressed by both CD4+CD25+ Tregs and IL-10 Tregs. Furthermore, suppression by these subsets is decreased in allergic individuals. In animal models, Tregs could be induced by high- or low-dose inhaled antigen, and prior induction of such Tregs prevented subsequent development of allergen sensitization and airway inflammation in inhaled challenge models. For many years, allergen-injection immunotherapy has been used for the therapy of allergic disease, and this treatment may induce IL-10 Tregs, leading to both suppression of Th2 responses and a switch from IgE to IgG4 antibody production. Improvements in allergen immunotherapy, such as peptide therapy, and greater understanding of the biology of Tregs hold great promise for the treatment and prevention of allergic disease.
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Affiliation(s)
- Douglas S Robinson
- Department of Allergy and Clinical Immunology, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London, United Kingdom
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38
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Abstract
For more than 90 years, allergen-specific immunotherapy, the only causative allergy treatment, has remained basically unchanged. The development of molecular biology techniques has led to the preparation of individual recombinant allergens from the most important allergen sources. Recombinant allergens can be used to determine the individual sensitization profile of allergic patients and have allowed the development of novel therapeutic tools. This article summarizes data on genetically modified recombinant allergen derivatives with reduced allergenic activity, which may be used to improve the safety and efficacy of specific immunotherapy.
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Affiliation(s)
- Verena Niederberger
- Department of Otorhinolaryngology, Vienna General Hospital, Medical University of Vienna, Ebene 8J, Waehringer Guertel 18-20, Vienna A-1090, Austria.
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39
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Abstract
In the past few decades, DNA technology has enabled the production of defined recombinant allergen molecules for diagnostic and therapeutic purposes. Recombinant allergens containing most of the relevant IgE epitopes present in natural allergen sources are now available and allergen proteins can be produced that are identical, without biological or batch-to-batch variation. A great advantage of recombinant allergens is that they can be used for component-resolved diagnostics, which makes it possible to establish the patient's individual IgE reactivity profile before therapy is selected. However, before recombinant allergens can be applied in clinical practice their biological activity has to be carefully investigated in vivo. We here describe the most commonly used provocation methods (skin tests (prick and intradermal), nasal, bronchial, and conjunctival provocations) and how they can be performed. We also discuss the results so far obtained with in vivo testing using recombinant allergens and envisage their future use for immunotherapy.
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Affiliation(s)
- M van Hage-Hamsten
- Department of Medicine, Division of Clinical Immunology and Allergy, Karolinska Institutet and Hospital, Stockholm, Sweden
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Niederberger V, Horak F, Vrtala S, Spitzauer S, Krauth MT, Valent P, Reisinger J, Pelzmann M, Hayek B, Kronqvist M, Gafvelin G, Grönlund H, Purohit A, Suck R, Fiebig H, Cromwell O, Pauli G, van Hage-Hamsten M, Valenta R. Vaccination with genetically engineered allergens prevents progression of allergic disease. Proc Natl Acad Sci U S A 2004; 101 Suppl 2:14677-82. [PMID: 15310844 PMCID: PMC521981 DOI: 10.1073/pnas.0404735101] [Citation(s) in RCA: 259] [Impact Index Per Article: 12.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
IgE-mediated allergy affects >25% of the population in industrialized countries. Repeated contact with the disease-eliciting allergens induces rises of allergen-specific IgE Abs and progression of the disease to more severe manifestations. Our study uses a type of vaccine that is based on genetically modified allergen derivatives to treat allergic patients. We developed hypoallergenic derivatives of the major birch pollen allergen, Bet v 1, by genetic engineering and vaccinated birch pollen-allergic patients (n = 124) in a double-blind, placebo-controlled study. Active treatment induced protective IgG Abs that inhibited allergen-induced release of inflammatory mediators. We also observed a reduction of cutaneous sensitivity as well as an improvement of symptoms in actively treated patients. Most important, rises of allergen-specific IgE induced by seasonal birch pollen exposure were significantly reduced in vaccinated patients. Vaccination with genetically engineered allergen derivatives is a therapy for allergy that not only ameliorates allergic reactions but also reduces the IgE production underlying the disease.
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Affiliation(s)
- V Niederberger
- Department of Otorhinolaryngology, Vienna General Hospital, University of Vienna, 1090 Vienna, Austria
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Vrtala S, Focke-Tejkl M, Swoboda I, Kraft D, Valenta R. Strategies for converting allergens into hypoallergenic vaccine candidates. Methods 2004; 32:313-20. [PMID: 14962766 DOI: 10.1016/j.ymeth.2003.08.016] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 08/21/2003] [Indexed: 11/27/2022] Open
Abstract
Specific immunotherapy is based on the administration of increasing doses of allergens to allergic patients with the aim of inducing a state of antigen-specific unresponsiveness. Specific immunotherapy is one of the few causative treatment approaches for Type I allergy but may cause numerous side effects, including local inflammatory reactions, systemic manifestations (e.g., asthma attacks) and in the worst case, anaphylactic shock which may lead to death. Several attempts have been made in the past to reduce the rate of side effects. They included the chemical modification of allergen extracts to reduce their allergenic activity and the adsorption of allergen extracts to adjuvants to prevent the systemic release of allergens after administration. During the last decade, cDNAs coding for the most relevant allergens have been isolated and the corresponding allergens have been produced as recombinant molecules. Using allergen-encoding cDNAs, the amino acid sequence of allergens or purified recombinant allergens several strategies can now be applied to produce allergen derivatives with reduced allergenic activity for allergy vaccination in a controlled and reproducible manner. Currently, allergen-encoding cDNAs are used to engineer recombinant hypoallergenic allergen derivatives. According to the amino acid sequences and experimental epitope mapping data, synthetic peptides representing T- or B-cell epitopes are produced and purified recombinant allergens are coupled to novel adjuvants for vaccine formulation. In this article, strategies for the production and evaluation of allergen derivatives with reduced allergenic activity for allergy vaccination are described. These new vaccines hold great promise to improve the current practice of allergen-specific immunotherapy and maybe also used for prophylactic vaccination in the future.
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Affiliation(s)
- Susanne Vrtala
- Department of Pathophysiology, University of Vienna, Vienna A-1090, Austria
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Valenta R, Ball T, Focke M, Linhart B, Mothes N, Niederberger V, Spitzauer S, Swoboda I, Vrtala S, Westritschnig K, Kraft D. Immunotherapy of allergic disease. Adv Immunol 2004; 82:105-53. [PMID: 14975256 DOI: 10.1016/s0065-2776(04)82003-0] [Citation(s) in RCA: 65] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023]
Affiliation(s)
- R Valenta
- Division of Immunopathology, Department of Pathophysiology, University of Vienna, Medical School, Austria
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Mahler V, Vrtala S, Kuss O, Diepgen TL, Suck R, Cromwell O, Fiebig H, Hartl A, Thalhamer J, Schuler G, Kraft D, Valenta R. Vaccines for birch pollen allergy based on genetically engineered hypoallergenic derivatives of the major birch pollen allergen, Bet v 1. Clin Exp Allergy 2004; 34:115-22. [PMID: 14720271 DOI: 10.1111/j.1365-2222.2004.01857.x] [Citation(s) in RCA: 35] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
BACKGROUND We have recently engineered recombinant derivatives of the major birch pollen allergen Bet v 1 (rBet v 1 fragments and trimer) with strongly reduced allergenic activity. OBJECTIVE The aim of this study was the in vivo characterization of potential allergy vaccines based on Al(OH)3-adsorbed genetically modified rBet v 1 derivatives in mice. METHODS BALB/c mice were immunized either with courses of nine injections of increasing doses of Al(OH)3-adsorbed rBet v 1 wild-type, rBet v 1 fragments, rBet v 1 trimer or Al(OH)3 alone in weekly intervals or with three high-dose injections applied in intervals of 3 weeks. Humoral immune responses to rBet v 1 wild-type and homologous plant allergens were measured by ELISA and Western blotting, and the ability of mouse antibodies to inhibit the binding of allergic patients IgE to Bet v 1 was studied by ELISA competition experiments. RESULTS In both schemes, hypoallergenic rBet v 1 derivatives induced low IgE but high IgG1 responses against rBet v 1 wild-type. The IgG1 antibodies induced by genetically modified rBet v 1 derivatives cross-reacted with natural Bet v 1 and its homologues from alder (Aln g 1) as well as hazel (Cor a 1) and strongly inhibited the binding of birch pollen allergic patients' IgE to Bet v 1 wild-type. CONCLUSION Genetically modified hypoallergenic rBet v 1 derivatives induce blocking antibodies in vivo. Their safety and efficacy for the treatment of birch pollen and associated plant allergies can now be evaluated in clinical immunotherapy studies.
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Affiliation(s)
- V Mahler
- Department of Dermatology, University of Erlangen, Germany
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Harwanegg C, Laffer S, Hiller R, Mueller MW, Kraft D, Spitzauer S, Valenta R. Microarrayed recombinant allergens for diagnosis of allergy. Clin Exp Allergy 2003; 33:7-13. [PMID: 12534543 DOI: 10.1046/j.1365-2222.2003.01550.x] [Citation(s) in RCA: 113] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
We suggest that the coapplication of recombinant allergens and microarray technology can lead to the development of new forms of multi-allergen tests which allow the determining and monitoring of complex sensitization profiles of allergic patients in single assays. The allergen extracts which have so far been used for diagnosis only allowed the determining of whether an allergic patient is sensitized against a particular allergen source, but the disease-eliciting allergens could not be identified. Through the application of recombinant DNA technology a rapidly growing panel of recombinant allergen molecules has become available which meanwhile comprises the epitope spectrum of most of the important allergen sources. We demonstrate that microarray technology can be used to establish multi-allergen tests consisting of microarrayed recombinant allergen molecules. Microarrayed recombinant allergens can be used to determine and monitor the profile of disease-eliciting allergens using single tests that require minute amounts of serum from allergic patients. The wealth of diagnostic information gained through microarray-based allergy testing will likely improve diagnosis, prevention and treatment of allergy.
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Affiliation(s)
- C Harwanegg
- VBC Genomics Bioscience Research GmbH, Vienna, Austria
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Abstract
During the past decade, genetic information for most of the common allergens has been obtained. Using these genetic blueprints it has become possible to reconstruct, by recombinant DNA technology, almost complete repertoires of the relevant allergens and their epitopes. Recombinant allergens with the allergenic features of naturally occurring allergens have promoted allergy research and form the basis of new multiallergen tests for refined allergy diagnosis. Allergen derivatives with reduced allergenic activity have also been produced by recombinant DNA technology to increase safety and specificity of allergen-specific immunotherapy. These derivatives can be engineered to contain relevant T cell epitopes and to maintain those sequence motifs which are required for inducing protective antibody responses and therefore hold great promise for improving allergen-specific immunotherapy.
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Affiliation(s)
- Rudolf Valenta
- Department of Pathophysiology, University of Vienna Medical School, Vienna General Hospital, Waehringer Guertel 18-20, A-1090, Vienna, Austria.
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van Hage-Hamsten M, Johansson E, Roquet A, Peterson C, Andersson M, Greiff L, Vrtala S, Valenta R, Grönneberg R. Nasal challenges with recombinant derivatives of the major birch pollen allergen Bet v 1 induce fewer symptoms and lower mediator release than rBet v 1 wild-type in patients with allergic rhinitis. Clin Exp Allergy 2002; 32:1448-53. [PMID: 12372124 DOI: 10.1046/j.1365-2745.2002.01495.x] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/04/2023]
Abstract
BACKGROUND Genetic engineering of the major birch pollen allergen (Bet v 1) has led to the generation of recombinant Bet v 1 derivatives with markedly reduced IgE-binding capacity, but with retained T cell activating ability. OBJECTIVE To compare the mucosal reactivity to rBet v 1 derivatives with rBet v 1 wild-type as basis for new therapeutic strategies for birch pollen allergy based on mucosal tolerance induction. METHODS Outside the pollen season, 10 patients with birch pollen allergic rhinitis and mild asthma underwent four nasal challenge-sessions in a randomized, double-blind, and cross-over design, employing increasing doses of rBet v 1 fragment mix, rBet v 1 trimer, rBet v 1 wild-type and diluent (albumin). Nasal lavage fluids (NAL) were collected before the challenge-series as well as 10 min, 4 and 24 h thereafter. Nasal lavage fluid levels of tryptase as well as EPO and ECP were measured as indices of mast cell and eosinophil activity, respectively. RESULTS All 10 patients tolerated the highest accumulated dose, 8.124 microg, when challenged with rBet v 1 trimer, eight with rBet v 1 fragments compared to one when challenged with rBet v 1 wild-type. No late phase reactions were observed. The change in tryptase levels (pre-challenge vs. 10 min) was significantly lower after challenges with rBet v 1 trimer and rBet v 1 fragments than with rBet v 1 wild-type. The change in EPO/ECP concentration pre-challenge versus 4 h post-challenge was lower for rBet v 1 trimer and the change was significantly lower when pre-challenge versus 24 h post-challenge to rBet v 1 fragments and rBet v 1 wild-type was examined. CONCLUSION The derivatives induced significantly fewer symptoms and lower mast cell and eosinophil activation than rBet v 1 wild-type upon application to the nasal mucosa. They could in the future be candidates for immunotherapy based on mucosal tolerance induction.
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Affiliation(s)
- M van Hage-Hamsten
- Department of Medicine, Division of Clinical Immunology, Karolinska Institute and Hospital, Stockholm, Sweden.
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Abstract
More than 25% of the population in industrialized countries suffers from immunoglobulin-E-mediated allergies. The antigen-specific immunotherapy that is in use at present involves the administration of allergen extracts to patients with the aim to cure allergic symptoms. However, the risk of therapy-induced side effects limits its broad application. Recent work indicates that the epitope complexity of natural allergen extracts can be recreated using recombinant allergens, and hypoallergenic derivatives of these can be engineered to increase treatment safety. It is proposed that these modified molecules will improve the current practice of specific immunotherapy and form a basis for prophylactic vaccination.
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Affiliation(s)
- Rudolf Valenta
- Department of Pathophysiology, University of Vienna Medical School, Vienna General Hospital-AKH, Australia.
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Pomés A, Chapman MD. Can knowledge of the molecular structure of allergens improve immunotherapy? Curr Opin Allergy Clin Immunol 2001; 1:549-54. [PMID: 11964740 DOI: 10.1097/00130832-200112000-00010] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
Conventional immunotherapy may be associated with the development of adverse reactions, including anaphylaxis, due to the use of increasing doses of allergen. Standardization of extracts is necessary in order to assess the correct amount of allergen administered. In recent years, increased knowledge on the molecular structure of allergens has allowed the development of novel alternatives for immunotherapy. Initially, allergens were cloned and expressed as recombinant proteins in eukaryotic and prokaryotic systems. Crystallization of the purified proteins led to the elucidation of the tertiary structure of the allergen. Molecular biology techniques were used to construct modified allergens whose new IgE binding properties were studied. IgE antibody mapping combined with molecular modeling has allowed the recognition of IgE binding sites on the surface of the molecule. This information has been applied to the engineering of new modified allergens, with and without adjuvants, that retain immunogenicity but with reduced allergenicity. The use of these molecules for immunotherapy should allow the administration of greater doses of allergen, without the undesired side effects characteristic of conventional immunotherapy.
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Affiliation(s)
- A Pomés
- Asthma and Allergic Diseases Center, Department of Medicine, UVA Health System, Charlottesville, Virginia, USA.
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Affiliation(s)
- C Grégoire
- Asthma and Allergic Diseases Center, Department of Medicine, University of Virginia, PO Box 801355, Charlottesville, VA 22908, USA
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