Bordas-Le Floch Véronique, Le Mignon Maxime, Bussières Laetitia, Jain Karine, Martelet Armelle, Baron-Bodo Véronique, Nony Emmanuel, Mascarell Laurent, Moingeon Philippe
Stallergenes Greer, Antony cedex, France.
PLoS One. 2017 Oct 5;12(10):e0185830. doi: 10.1371/journal.pone.0185830. eCollection 2017.
House dust mites (HDMs) such as Dermatophagoides farinae and D. pteronyssinus represent major causes of perennial allergy. HDM proteomes are currently poorly characterized, with information mostly restricted to allergens. As of today, 33 distinct allergen groups have been identified for these 2 mite species, with groups 1 and 2 established as major allergens. Given the multiplicity of IgE-reactive mite proteins, potential additional allergens have likely been overlooked.
To perform a comprehensive characterization of the transcriptomes, proteomes and allergomes of D. farinae and D. pteronyssinus in order to identify novel allergens.
Transcriptomes were analyzed by RNA sequencing and de novo assembly. Comprehensive mass spectrometry-based analyses proteomes were combined with two-dimensional IgE reactivity profiling.
Transcripts from D. farinae and D. pteronyssinus were assembled, translated into protein sequences and used to populate derived sequence databases in order to inform immunoproteomic analyses. A total of 527 and 157 proteins were identified by bottom-up MS analyses in aqueous extracts from purified HDM bodies and fecal pellets, respectively. Based on high sequence similarities (>71% identity), we also identified 2 partial and 11 complete putative sequences of currently undisclosed D. pteronyssinus counterparts of D. farinae registered allergens. Immunoprofiling on 2D-gels revealed the presence of unknown 23 kDa IgE reactive proteins in both species. Following expression of non-glycosylated recombinant forms of these molecules, we confirm that these new allergens react with serum IgEs from 42% (8/19) of HDM-allergic individuals.
Using combined transcriptome and immunoproteome approaches, we provide a comprehensive characterization of D. farinae and D. pteronyssinus allergomes. We expanded the known allergen repertoire for D. pteronyssinus and identified two novel HDM allergens, now officially referred by the International Union of Immunological Societies (IUIS) Nomenclature Subcommittee as Der f 36 and Der p 36.
屋尘螨，如粉尘螨和户尘螨，是常年性过敏的主要诱因。目前对屋尘螨蛋白质组的特征了解甚少，相关信息大多局限于过敏原。截至目前，已为这两种螨类鉴定出33个不同的过敏原组，其中第1组和第2组被确定为主要过敏原。鉴于与IgE反应的螨蛋白种类繁多，可能还有其他潜在过敏原被忽视了。
对粉尘螨和户尘螨的转录组、蛋白质组和过敏原组进行全面表征，以鉴定新的过敏原。
通过RNA测序和从头组装分析转录组。基于质谱的综合分析将蛋白质组与二维IgE反应谱分析相结合。
对粉尘螨和户尘螨的转录本进行组装，翻译成蛋白质序列，并用于构建衍生序列数据库，以指导免疫蛋白质组分析。通过自下而上的质谱分析，在纯化的屋尘螨虫体和粪便颗粒的水提取物中分别鉴定出527种和157种蛋白质。基于高度的序列相似性（>71%同一性），我们还鉴定出了粉尘螨已注册过敏原的户尘螨对应物的2个部分和11个完整的推定序列。二维凝胶免疫分析显示这两种螨类中均存在未知的23 kDa IgE反应性蛋白。在表达这些分子的非糖基化重组形式后，我们证实这些新过敏原与42%（8/19）的屋尘螨过敏个体的血清IgE发生反应。
通过结合转录组和免疫蛋白质组方法，我们对粉尘螨和户尘螨的过敏原组进行了全面表征。我们扩展了户尘螨已知的过敏原库，并鉴定出两种新的屋尘螨过敏原，现已被国际免疫学会联盟（IUIS）命名小组委员会正式命名为Der f 36和Der p 36。
