Lactobacillus plantarum-derived Extracellular Vesicles Protect Atopic Dermatitis Induced by Staphylococcus aureus-derived Extracellular Vesicles - PubMed (original) (raw)

. 2018 Sep;10(5):516-532.

doi: 10.4168/aair.2018.10.5.516.

Seng Jin Choi # 2, Hyun Il Choi # 3, Jun Pyo Choi 4, Han Ki Park 5, Eun Kyoung Kim 6, Min Jeong Kim 7, Byoung Seok Moon 7, Taek Ki Min 8, Mina Rho 9, Young Joo Cho 10, Sanghwa Yang 6, Yoon Keun Kim 6, You Young Kim 11, Bok Yang Pyun 12

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Lactobacillus plantarum-derived Extracellular Vesicles Protect Atopic Dermatitis Induced by Staphylococcus aureus-derived Extracellular Vesicles

Min Hye Kim et al. Allergy Asthma Immunol Res. 2018 Sep.

Abstract

Purpose: The microbial environment is an important factor that contributes to the pathogenesis of atopic dermatitis (AD). Recently, it was revealed that not only bacteria itself but also extracellular vesicles (EVs) secreted from bacteria affect the allergic inflammation process. However, almost all research carried out so far was related to local microorganisms, not the systemic microbial distribution. We aimed to compare the bacterial EV composition between AD patients and healthy subjects and to experimentally find out the beneficial effect of some bacterial EV composition.

Methods: Twenty-seven AD patients and 6 healthy control subjects were enrolled. After urine and serum were obtained, EVs were prepared from samples. Metagenomic analysis of 16s ribosomal DNA extracted from the EVs was performed, and bacteria showing the greatest difference between controls and patients were identified. In vitro and in vivo therapeutic effects of significant bacterial EV were evaluated with keratinocytes and with Staphylococcus aureus-induced mouse AD models, respectively.

Results: The proportions of Lactococcus, Leuconostoc and Lactobacillus EVs were significantly higher and those of Alicyclobacillus and Propionibacterium were lower in the control group than in the AD patient group. Therefore, lactic acid bacteria were considered to be important ones that contribute to the difference between the patient and control groups. In vitro, interleukin (IL)-6 from keratinocytes and macrophages decreased and cell viability was restored with Lactobacillus plantarum-derived EV treatment prior to S. aureus EV treatment. In S. aureus-induced mouse AD models, L. plantarum-derived EV administration reduced epidermal thickening and the IL-4 level.

Conclusions: We suggested the protective role of lactic acid bacteria in AD based on metagenomic analysis. Experimental findings further suggest that L. plantarum-derived EV could help prevent skin inflammation.

Keywords: Atopic dermatitis; Lactobacillus; metagenomics; microbiome; probiotics.

Copyright © 2018 The Korean Academy of Asthma, Allergy and Clinical Immunology · The Korean Academy of Pediatric Allergy and Respiratory Disease.

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Conflict of interest statement

There are no financial or other issues that might lead to conflict of interest.

Figures

Fig. 1

Fig. 1. (A) Alpha-diversity as determined by the Chao 1 index. (B) Beta-diversity defined by PCA. (C) Bacterial community analysis of the urine of AD patients and normal controls at the phylum level. (D) Bacterial community analysis of the urine of patients and controls at the genus level (case: AD patients, control: healthy controls).

PCA, principal component analysis; AD, atopic dermatitis.

Fig. 2

Fig. 2. (A) Differences in bacterial EVs composition in the urine of AD patients and normal controls. (B) Bacterial community analysis of the urine of AD patients and normal controls at the genus level.

EV, extracellular vesicle; AD, atopic dermatitis.

Fig. 3

Fig. 3. Characterization of EVs from Lactobacillus plantarum. (A) TEM images of purified EVs from L. plantarum. (B) The size distribution of EVs from L. plantarum. The size distribution of EVs was measured in a diameter range of 20 to 50 nm. (C) SDS-PAGE analysis of purified proteins (WCL, EVs) from L. plantarum. Standard markers are shown on the left (kDa).

EV, extracellular vesicle; TEM, transmission electron microscopy; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; WCL, whole cell lysate.

Fig. 4

Fig. 4. (A) In vitro effects of Lactobacillus plantarum on the secretion of IL-6 from keratinocytes stimulated by Sa EV. NC was treated only with PBS, and PC was treated only with Sa EV. Pre- and co-treatment indicates that Lp EV was treated before Sa EV stimulation, or at the same time, respectively. (B) Keratinocytes cell viability assessment.

IL, interleukin; Sa EV, Staphylococcus aureus_-induced extracellular vesicle; NC, negative control; PBS, phosphate-buffered saline; PC, positive control; Lp EV, Lactobacillus plantarum_-derived extracellular vesicle. *P < 0.05 vs. NC; †_P < 0.05 vs. PC; ‡_P < 0.01 vs. PC (Mann-Whitney U test).

Fig. 5

Fig. 5. In vivo effects of oral Lp EV on the AD-like skin inflammation. (A) AD mouse model using tape-stripping and Sa EV. (B) Images of H&E-stained skin. (C) Histological analysis of epidermal thickness and infiltrating eosinophil. (D) Levels of IL-4 and IL-5.

Lp EV, Lactobacillus plantarum_-derived extracellular vesicle; AD, atopic dermatitis; Sa EV, Staphylococcus aureus_-induced extracellular vesicle; H&E, hematoxylin and eosin; IL, interleukin; NC, negative control; PC, positive control. *P < 0.05 vs. NC; †_P < 0.05 vs. PC; ‡_P < 0.01 vs. PC (Mann-Whitney U test).

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