School of Life Sciences, Chinese University of Hong Kong, Hong Kong, China.
State Key Laboratory of Respiratory Disease for Allergy at Shenzhen University, Shenzhen, China.
J Allergy Clin Immunol. 2015 Feb;135(2):539-48. doi: 10.1016/j.jaci.2014.09.031. Epub 2014 Oct 31.
A sequenced house dust mite (HDM) genome would advance our understanding of HDM allergens, a common cause of human allergies.
We sought to produce an annotated Dermatophagoides farinae draft genome and develop a combined genomic-transcriptomic-proteomic approach for elucidation of HDM allergens.
A D farinae draft genome and transcriptome were assembled with high-throughput sequencing, accommodating microbiome sequences. The allergen gene structures were validated by means of Sanger sequencing. The mite's microbiome composition was determined, and the predominant genus was validated immunohistochemically. The allergenicity of a ubiquinol-cytochrome c reductase binding protein homologue was evaluated with immunoblotting, immunosorbent assays, and skin prick tests.
The full gene structures of 20 canonical allergens and 7 noncanonical allergen homologues were produced. A novel major allergen, ubiquinol-cytochrome c reductase binding protein-like protein, was found and designated Der f 24. All 40 sera samples from patients with mite allergy had IgE antibodies against rDer f 24. Of 10 patients tested, 5 had positive skin reactions. The predominant bacterial genus among 100 identified species was Enterobacter (63.4%). An intron was found in the 13.8-kDa D farinae bacteriolytic enzyme gene, indicating that it is of HDM origin. The Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed a phototransduction pathway in D farinae, as well as thiamine and amino acid synthesis pathways, which is suggestive of an endosymbiotic relationship between D farinae and its microbiome.
An HDM genome draft produced from genomic, transcriptomic, and proteomic experiments revealed allergen genes and a diverse endosymbiotic microbiome, providing a tool for further identification and characterization of HDM allergens and development of diagnostics and immunotherapeutic vaccines.
序列分析的屋尘螨（HDM）基因组将有助于我们理解 HDM 过敏原，这是人类过敏的常见原因。
我们试图生成一个注释的粉尘螨（Dermatophagoides farinae）基因组草案，并开发一种组合基因组-转录组-蛋白质组方法来阐明 HDM 过敏原。
使用高通量测序组装 D farinae 基因组和转录组，同时容纳微生物组序列。通过 Sanger 测序验证过敏原基因结构。确定螨的微生物组组成，并通过免疫组织化学验证主要属。使用免疫印迹、免疫吸附测定和皮肤点刺试验评估泛醌-细胞色素 c 还原酶结合蛋白同源物的变应原性。
生成了 20 个经典过敏原和 7 个非经典过敏原同源物的完整基因结构。发现并命名了一种新的主要过敏原，泛醌-细胞色素 c 还原酶结合蛋白样蛋白，命名为 Der f 24。40 份尘螨过敏患者的血清样本均针对 rDer f 24 产生 IgE 抗体。10 名测试患者中有 5 名出现阳性皮肤反应。在鉴定的 100 个物种中，主要细菌属为肠杆菌（63.4%）。在 13.8 kDa D farinae 溶菌酶基因中发现一个内含子，表明它来自 HDM。京都基因与基因组百科全书通路分析显示 D farinae 存在光转导通路以及硫胺素和氨基酸合成通路，提示 D farinae 与其微生物组之间存在共生关系。
从基因组、转录组和蛋白质组实验生成的 HDM 基因组草案揭示了过敏原基因和多样化的共生微生物组，为进一步鉴定和表征 HDM 过敏原以及开发诊断和免疫治疗疫苗提供了工具。
