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The draft genome, transcriptome, and microbiome of Dermatophagoides farinae reveal a broad spectrum of dust mite allergens - 05/02/15

Doi : 10.1016/j.jaci.2014.09.031 
Ting-Fung Chan, PhD a, , Kun-Mei Ji, PhD b, , Aldrin Kay-Yuen Yim, BS a, c, , Xiao-Yu Liu, MSc d, , Jun-Wei Zhou, PhD e, , , Rui-Qi Li, BSM d, , Kevin Yi Yang, PhD c, e, , Jing Li, MS f, , Meng Li, MSc d, Patrick Tik-Wan Law, PhD a, Yu-Lan Wu, BS d, Ze-Lang Cai, MSc d, Hao Qin, PhD a, c, Ying Bao, MSc d, Ross Ka-Kit Leung, PhD c, e, Patrick Kwok-Shing Ng, PhD e, Ju Zou, MSc d, Xiao-Jun Zhong, MSc d, Pi-Xin Ran, MD f, Nan-Shan Zhong, MSc f, , Zhi-Gang Liu, MD b, d, , Stephen Kwok-Wing Tsui, PhD c, e,
a School of Life Sciences, Chinese University of Hong Kong, Hong Kong, China 
c Hong Kong Bioinformatics Centre, Chinese University of Hong Kong, Hong Kong, China 
e School of Biomedical Sciences, Chinese University of Hong Kong, Hong Kong, China 
b State Key Laboratory of Respiratory Disease for Allergy at Shenzhen University, Shenzhen, China 
d Shenzhen Key Laboratory of Allergy and Immunology, School of Medicine, Shenzhen University, Shenzhen, China 
f State Key Laboratory of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China 

Corresponding author: Nan-Shan Zhong, MSc, State Key Laboratory of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical College No. 151, Yanjiang Rd, Guangzhou 510120, China.∗∗Zhi-Gang Liu, MD, State Key Laboratory of Respiratory Disease for Allergy at Shenzhen University and Shenzhen Key Laboratory of Allergy and Immunology, School of Medicine, Shenzhen University, Nanhai Ave 3688, 518060 Shenzhen, Guangdong, People's Republic of China.∗∗∗Stephen Kwok-Wing Tsui, PhD, School of Biomedical Sciences, the Chinese University of Hong Kong, Shatin, N.T., Hong Kong.

Abstract

Background

A sequenced house dust mite (HDM) genome would advance our understanding of HDM allergens, a common cause of human allergies.

Objective

We sought to produce an annotated Dermatophagoides farinae draft genome and develop a combined genomic-transcriptomic-proteomic approach for elucidation of HDM allergens.

Methods

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.

Results

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.

Conclusion

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.

Il testo completo di questo articolo è disponibile in PDF.

Key words : House dust mite, allergen, genome, microbiome, transcriptome, proteome, ubiquinol-cytochrome c reductase binding protein, Der f 24, Enterobacter species

Abbreviations used : CEG, CEGMA, Gb, GO, HDM, KEGG, NCBI, UQCRB


Mappa


 Supported by the NSFC (81071388; 31328014; 3027122), Guangdong-Hong Kong Technology Cooperation Foundation (2005A10905007), the China 863 (2006AA02A231) and 973 (2009CB522100) Project Foundations, the Chinese University of Hong Kong Focused Investment Funding, the Hong Kong Research Grants Council (475113), the Shenzhen Science Foundation (JCYJ20120613113021045; JCYJ20130326112225593) and the Guangdong Province NSF (no. 2014566).
 Disclosure of potential conflict of interest: The authors declare that they have no relevant conflicts of interest.


© 2014  The Authors. Pubblicato da Elsevier Masson SAS. Tutti i diritti riservati.
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