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Over the last four decades, molecular cloning has evolved tremendously. Efficient products allowing assembly of multiple DNA fragments have become available. However, cost-effective tools for engineering antibodies of different specificities, isotypes and species are still needed for many research and clinical applications in academia. Here, we report a(More)
BACKGROUND Grass pollen immunotherapy is an effective treatment for seasonal allergic rhinitis that provides the opportunity to study the induction and maintenance of allergen-specific immune tolerance. OBJECTIVES We investigated the relationship between clinical responsiveness, regulatory cytokine production, and antibody responses to allergen during 1(More)
The incidence of allergic diseases is increasing at an alarming rate, particularly in countries with a western lifestyle. Currently in the UK, approximately one quarter of the population suffers from seasonal allergic rhinitis. Most patients can be treated with conventional pharmacotherapy on an 'as needed' symptomatic basis whereas allergen immunotherapy(More)
BACKGROUND Grass pollen immunotherapy for allergic rhinitis is a disease-modifying treatment that results in long-term clinical tolerance lasting years after treatment discontinuation. Active treatment is associated with generation of inhibitory grass pollen-specific IgG antibodies capable of blocking allergen-IgE interactions. OBJECTIVES We sought to(More)
BACKGROUND Serum IgG(4) responses to allergen immunotherapy are well documented as blocking allergen binding to receptor-bound IgE on antigen-presenting cells and effector cells, but the molecular characteristics of treatment-induced IgG(4), particularly in relation to expressed antibody, are poorly defined. OBJECTIVES We aimed to clone and express(More)
BACKGROUND Most children with detectable peanut-specific IgE (P-sIgE) are not allergic to peanut. We addressed 2 non-mutually exclusive hypotheses for the discrepancy between allergy and sensitization: (1) differences in P-sIgE levels between children with peanut allergy (PA) and peanut-sensitized but tolerant (PS) children and (2) the presence of an IgE(More)
Allergen immunotherapy is an effective treatment option in patients with IgE-mediated allergy, particularly in those who do not respond to usual antiallergic drugs. Successful immunotherapy has been associated with the suppression of allergic inflammation in target organs and increases in allergen-specific IgG antibodies, particularly the IgG(4) subclass.(More)
IgG4 purified from patients undergoing specific allergen immunotherapy inhibits the activities of the serum IgE in in vitro assays and is thought to reduce the symptoms of the disease. However, it is not known whether this is related to an intrinsic property of this subclass or only the allergen specificity. We tested the hypothesis that allergen(More)
BACKGROUND Previous studies of immunoglobulin gene sequences in patients with allergic diseases using low-throughput Sanger sequencing have limited the analytic depth for characterization of IgE repertoires. OBJECTIVES We used a high-throughput, next-generation sequencing approach to characterize immunoglobulin heavy-chain gene (IGH) repertoires in(More)
BACKGROUND The calcium-binding 2EF-hand protein Phl p 7 from timothy grass pollen is a highly cross-reactive pollen pan-allergen that can induce severe clinical symptoms in allergic patients. Recently, a human monoclonal Phl p 7-specific IgG4 antibody (mAb102.1F10) was isolated from a patient who had received grass pollen-specific immunotherapy (SIT). (More)