Abstract
BackgroundAllergy to short ragweed (Ambrosia artemisiifolia) pollen is a serious and expanding health problem in North America and Europe. Whereas only 10 short ragweed pollen allergens are officially recorded, patterns of IgE reactivity observed in ragweed allergic patients suggest that other allergens contribute to allergenicity. The objective of the present study was to identify novel allergens following extensive characterization of the transcriptome and proteome of short ragweed pollen.MethodsFollowing a Proteomics-Informed-by-Transcriptomics approach, a comprehensive transcriptomic data set was built up from RNA-seq analysis of short ragweed pollen. Mass spectrometry-based proteomic analyses and IgE reactivity profiling after high resolution 2D-gel electrophoresis were then combined to identify novel allergens.ResultsShort ragweed pollen transcripts were assembled after deep RNA sequencing and used to inform proteomic analyses, thus leading to the identification of 573 proteins in the short ragweed pollen. Patterns of IgE reactivity of individual sera from 22 allergic patients were assessed using an aqueous short ragweed pollen extract resolved over 2D-gels. Combined with information derived from the annotated pollen proteome, those analyses revealed the presence of multiple unreported IgE reactive proteins, including new Amb a 1 and Amb a 3 isoallergens as well as 7 novel candidate allergens reacting with IgEs from 20–70% of patients. The latter encompass members of the carbonic anhydrase, enolase, galactose oxidase, GDP dissociation inhibitor, pathogenesis related-17, polygalacturonase and UDP-glucose pyrophosphorylase families.ConclusionsWe extended the list of allergens identified in short ragweed pollen. These findings have implications for both diagnosis and allergen immunotherapy purposes.
Highlights
Exposure to short ragweed (Ambrosia artemisiifolia) pollen is a major cause of severe type I respiratory allergy [1]
Whereas only 10 short ragweed pollen allergens are officially recorded, patterns of IgE reactivity observed in ragweed allergic patients suggest that other allergens contribute to allergenicity
Combined with information derived from the annotated pollen proteome, those analyses revealed the presence of multiple unreported IgE reactive proteins, including new Amb a 1 and Amb a 3 isoallergens as well as 7 novel candidate allergens reacting with IgEs from 20–70% of patients
Summary
Exposure to short ragweed (Ambrosia artemisiifolia) pollen is a major cause of severe type I respiratory allergy [1]. 10 short ragweed pollen allergens have been identified and recorded by the International Union of Immunological Societies (IUIS). This includes the pectate lyase Amb a 1, considered as the major short ragweed pollen allergen with approximately 90% of ragweed-allergic individuals exhibiting IgE reactivity to this molecule [4,5,6]. We recently identified as a new major allergen a cysteine protease reactive with seric IgEs from 66% of ragweed-allergic patients, recorded as Amb a 11 by the IUIS [11]. Allergy to short ragweed (Ambrosia artemisiifolia) pollen is a serious and expanding health problem in North America and Europe. The objective of the present study was to identify novel allergens following extensive characterization of the transcriptome and proteome of short ragweed pollen
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