A Metagenomic Analysis Provides a Culture-Independent Pathogen Detection for Atopic Dermatitis

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Abstract

Purpose

Atopic dermatitis (AD) is an inflammatory skin disease, significantly affecting the quality of life. Using AD as a model system, we tested a successive identification of AD-associated microbes, followed by a culture-independent serum detection of the identified microbe.

Methods

A total of 43 genomic DNA preparations from washing fluid of the cubital fossa of 6 healthy controls, skin lesions of 27 AD patients, 10 of which later received treatment (post-treatment), were subjected to high-throughput pyrosequencing on a Roche 454 GS-FLX platform.

Results

Microbial diversity was decreased in AD, and was restored following treatment. AD was characterized by the domination of Staphylococcus, Pseudomonas, and Streptococcus, whereas Alcaligenaceae (f), Sediminibacterium, and Lactococcus were characteristic of healthy skin. An enzyme-linked immunosorbent assay (ELISA) showed that serum could be used as a source for the detection of Staphylococcus aureus extracellular vesicles (EVs). S. aureus EV-specific immunoglobulin G (IgG) and immunoglobulin E (IgE) were quantified in the serum.

Conclusions

A metagenomic analysis together with a serum detection of pathogen-specific EVs provides a model for successive identification and diagnosis of pathogens of AD.

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Most cited references 19

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New insights into atopic dermatitis.

Donald Y M Leung, Mark Boguniewicz, Michael Howell … (2004)

Atopic dermatitis is a chronic inflammatory skin disease associated with cutaneous hyperreactivity to environmental triggers and is often the first step in the atopic march that results in asthma and allergic rhinitis. The clinical phenotype that characterizes atopic dermatitis is the product of interactions between susceptibility genes, the environment, defective skin barrier function, and immunologic responses. This review summarizes recent progress in our understanding of the pathophysiology of atopic dermatitis and the implications for new management strategies.

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Severity scoring of atopic dermatitis: the SCORAD index. Consensus Report of the European Task Force on Atopic Dermatitis.

A-M Calza, R Lever, DJ Atherton … (1993)

Assessment methods for atopic dermatitis (AD) are not standardized, and therapeutic studies are difficult to interpret. To obtain a consensus on assessment methods in AD and to use a statistical method to develop a composite severity index. Consensus definitions were given for items used in the scoring system (extent, intensity, subjective) and illustrated for intensity items. Slides were reviewed to address within- and between-observer variability by a group of 10 trained clinicians, and data were statistically evaluated with a two-way analysis of variance. Two variants of an assessment system were compared in 88 patients at 5 different institutions. Data were analyzed using principal-component analysis. For 5 intensity items studied (erythema, edema/papulation, oozing/crusts, excoriations, lichenification), within- and between-observer variability was good overall, except for edema/papulation which was difficult to assess with slides. In the series of 88 patients, principal-component analysis allowed to extract two unrelated components: the first one accounting for 33% of total variance was interpreted as a 'severity' component; the second one, accounting for 18% of variance, was interpreted as a 'profile' component distinguishing patients with mostly erythema and subjective symptoms and those with mostly lichenification and dryness and lower subjective symptoms. Of the two evaluation systems used, the one using the rule of nine to assess extent was found more workable than the one using a distribution x intensity product. A scoring index (SCORAD) combining extent, severity and subjective symptoms was mathematically derived from the first system and showed a normal distribution of the population studied. The final choice for the evaluation system was mostly made based on simplicity and easy routine use in outpatient clinics. Based on mathematical appreciation of weights of the items used in the assessment of AD, extent and subjective symptoms account for around 20% each of the total score, intensity items representing 60%. The so-designed composite index SCORAD needs to be further tested in clinical trials.

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Extracellular vesicles derived from Staphylococcus aureus induce atopic dermatitis-like skin inflammation

S Hong, M Kim, E-Y Lee … (2011)

Background Recently, we found that Staphylococcus aureus produces extracellular vesicles (EV) that contain pathogenic proteins. Although S. aureus infection has been linked with atopic dermatitis (AD), the identities of the causative agents from S. aureus are controversial. We evaluated whether S. aureus-derived EV are causally related to the pathogenesis of AD. Methods Extracellular vesicles were isolated by the ultracentrifugation of S. aureus culture media. The EV were applied three times per week to tape-stripped mouse skin. Inflammation and immune dysfunction were evaluated 48 h after the final application in hairless mice. Extracellular vesicles-specific IgE levels were measured by ELISA in AD patients and healthy subjects. Results The in vitro application of S. aureus EV increased the production of pro-inflammatory mediators (IL-6, thymic stromal lymphopoietin, macrophage inflammatory protein-1α, and eotaxin) by dermal fibroblasts. The in vivo application of S. aureus EV after tape stripping caused epidermal thickening with infiltration of the dermis by mast cells and eosinophils in mice. These changes were associated with the enhanced cutaneous production of IL-4, IL-5, IFN-γ, and IL-17. Interestingly, the serum levels of S. aureus EV-specific IgE were significantly increased in AD patients relative to healthy subjects. Conclusion These results indicate that S. aureus EV induce AD-like inflammation in the skin and that S. aureus-derived EV are a novel diagnostic and therapeutic target for the control of AD.

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Author and article information

Journal

Journal ID (nlm-ta): Allergy Asthma Immunol Res

Journal ID (iso-abbrev): Allergy Asthma Immunol Res

Journal ID (publisher-id): AAIR

Title: Allergy, Asthma & Immunology Research

Publisher: The Korean Academy of Asthma, Allergy and Clinical Immunology; The Korean Academy of Pediatric Allergy and Respiratory Disease

ISSN (Print): 2092-7355

ISSN (Electronic): 2092-7363

Publication date (Print): September 2017

Publication date (Electronic): 07 June 2017

Volume: 9

Issue: 5

Pages: 453-461

Affiliations

[1 ]Department of Internal Medicine, Ewha Womans University School of Medicine, Seoul, Korea.

[2 ]Department of Computer Science and Engineering, Hanyang University, Seoul, Korea.

[3 ]Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.

[4 ]Division of Molecular and Life Sciences, Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.

[5 ]Department of Allergy and Clinical Immunology, Ajou University Medical Center, Suwon, Korea.

[6 ]Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea.

[7 ]Department of Pediatrics, Pediatric Allergy and Respiratory Center, Soonchunhyang University Hospital, Soonchunhyang University College of Medicine, Seoul, Korea.

[8 ]Institute of MD Healthcare, Seoul, Korea.

Author notes

Correspondence to: Bok Yang Pyun, MD, Department of Pediatrics, Pediatric Allergy and Respiratory Center, Soonchunhyang University Hospital, Soonchunhyang University College of Medicine, 59 Daesagwan-ro, Yongsan-gu, Seoul 04401, Korea. Tel: +82-2-709-9339; Fax: +82-2-794-5471; bypyun@ 123456schmc.ac.kr

Co-Correspondence to: Sanghwa Yang, PhD, Institute of MD Healthcare, 9 World Cup buk-ro 56-gil, Mapo-gu, Seoul 03923, Korea. Tel: +82-70-7812-8061; Fax: +82-2-2655-0768; arraychip@ 123456gmail.com

Min-Hye Kim and Mina Rho contributed equally to this paper.

Article

DOI: 10.4168/aair.2017.9.5.453

PMC ID: 5500701

PubMed ID: 28677360

SO-VID: 112af74b-98bd-469b-a675-af19f15f64e4

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

History

Date received : 14 November 2016

Date revision received : 27 March 2017

Date accepted : 04 April 2017

Funding

Funded by: Ministry of Health and Welfare, CrossRef http://dx.doi.org/10.13039/501100003625;

Award ID: HI13C0040

Award ID: HI14C2628

Funded by: National Research Foundation of Korea, CrossRef http://dx.doi.org/10.13039/501100003725;

Award ID: NRF-2010-0028684

Award ID: NRF-2014-R1A1A1005144

Funded by: Soonchunhyang University, CrossRef http://dx.doi.org/10.13039/501100002560;

A Metagenomic Analysis Provides a Culture-Independent Pathogen Detection for Atopic Dermatitis

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Abstract

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Conclusions

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Karger: Dermatology

Most cited references 19

New insights into atopic dermatitis.

Severity scoring of atopic dermatitis: the SCORAD index. Consensus Report of the European Task Force on Atopic Dermatitis.

Extracellular vesicles derived from Staphylococcus aureus induce atopic dermatitis-like skin inflammation

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