Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>

<i>Helicobacter pylori</i> is a pathogenic bacterium that causes gastric and extragastric diseases. We have previously demonstrated that one of the mechanisms of <i>H. pylori</i>-associated chronic immune thrombocytopenia involves immune complexes of platelets, a <i>H....

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Main Authors: Aoi Okamoto, Tatsuki Shibuta, Nanaka Morita, Ryota Fujinuma, Masaya Shiraishi, Reimi Matsuda, Mayu Okada, Satoe Watanabe, Tsukuru Umemura, Hiroaki Takeuchi
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Language:English
Published: MDPI AG 2025-03-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/4/753
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author Aoi Okamoto
Tatsuki Shibuta
Nanaka Morita
Ryota Fujinuma
Masaya Shiraishi
Reimi Matsuda
Mayu Okada
Satoe Watanabe
Tsukuru Umemura
Hiroaki Takeuchi
author_facet Aoi Okamoto
Tatsuki Shibuta
Nanaka Morita
Ryota Fujinuma
Masaya Shiraishi
Reimi Matsuda
Mayu Okada
Satoe Watanabe
Tsukuru Umemura
Hiroaki Takeuchi
author_sort Aoi Okamoto
collection DOAJ
description <i>Helicobacter pylori</i> is a pathogenic bacterium that causes gastric and extragastric diseases. We have previously demonstrated that one of the mechanisms of <i>H. pylori</i>-associated chronic immune thrombocytopenia involves immune complexes of platelets, a <i>H. pylori</i> protein Lpp20 and an anti-Lpp20 antibody. However, it remains unclear how Lpp20 enters the body. We hypothesize that bacterial extracellular vesicles (bEVs) transport Lpp20. Thus, this study assessed Lpp20 in the bEVs released from seven clinical <i>H. pylori</i> isolates, using immunoprecipitation (IP), immunoblotting (IB), and surface plasmon resonance imaging (SPRi), with anti-GroEL (a marker of bEVs) and anti-Lpp20 antibodies. Lpp20 and bEVs were each detected in lysates of all seven strains. IP–IB experiments demonstrated that bEVs containing Lpp20 were produced by five of the strains (J99, SS1, HPK5, JSHR3, and JSHR31). SPRi using an anti-Lpp20 antibody demonstrated significantly higher reflectance from the strain HPK5 than from its <i>lpp20</i>-disrupted strains (<i>p</i> < 0.01), indicating localization of Lpp20 on the bEVs’ surface; Lpp20 may also be contained within bEVs. The bEVs containing Lpp20 were not detected from two clinical <i>H. pylori</i> strains (26695 and JSHR6) or from two <i>lpp20</i>-disrupted strains (26695ΔLpp20 and HPK5ΔLpp20). Differences in Lpp20 detection in bEVs are likely due to variations in bEV production resulting from strain diversity.
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spelling doaj-art-1269bbddbc3f42fb9ed40a44418cb2bb2025-08-20T02:28:37ZengMDPI AGMicroorganisms2076-26072025-03-0113475310.3390/microorganisms13040753Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>Aoi Okamoto0Tatsuki Shibuta1Nanaka Morita2Ryota Fujinuma3Masaya Shiraishi4Reimi Matsuda5Mayu Okada6Satoe Watanabe7Tsukuru Umemura8Hiroaki Takeuchi9Medical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanDepartment of Medical Science Technology, School of Health Science at Fukuoka, International University of Health and Welfare, 137-1 Enokiz, Okawa 831-8501, JapanDepartment of Medical Science Technology, School of Health Science at Narita, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanDepartment of Medical Science Technology, School of Health Science at Narita, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanMedical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanMedical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanMedical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanDepartment of Medical Science Technology, School of Health Science at Narita, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, JapanDepartment of Medical Science Technology, School of Health Science at Fukuoka, International University of Health and Welfare, 137-1 Enokiz, Okawa 831-8501, JapanMedical Laboratory Science, Graduate School of Health and Welfare Sciences, International University of Health and Welfare, 4-3 Kozunomori, Narita 286-8686, Japan<i>Helicobacter pylori</i> is a pathogenic bacterium that causes gastric and extragastric diseases. We have previously demonstrated that one of the mechanisms of <i>H. pylori</i>-associated chronic immune thrombocytopenia involves immune complexes of platelets, a <i>H. pylori</i> protein Lpp20 and an anti-Lpp20 antibody. However, it remains unclear how Lpp20 enters the body. We hypothesize that bacterial extracellular vesicles (bEVs) transport Lpp20. Thus, this study assessed Lpp20 in the bEVs released from seven clinical <i>H. pylori</i> isolates, using immunoprecipitation (IP), immunoblotting (IB), and surface plasmon resonance imaging (SPRi), with anti-GroEL (a marker of bEVs) and anti-Lpp20 antibodies. Lpp20 and bEVs were each detected in lysates of all seven strains. IP–IB experiments demonstrated that bEVs containing Lpp20 were produced by five of the strains (J99, SS1, HPK5, JSHR3, and JSHR31). SPRi using an anti-Lpp20 antibody demonstrated significantly higher reflectance from the strain HPK5 than from its <i>lpp20</i>-disrupted strains (<i>p</i> < 0.01), indicating localization of Lpp20 on the bEVs’ surface; Lpp20 may also be contained within bEVs. The bEVs containing Lpp20 were not detected from two clinical <i>H. pylori</i> strains (26695 and JSHR6) or from two <i>lpp20</i>-disrupted strains (26695ΔLpp20 and HPK5ΔLpp20). Differences in Lpp20 detection in bEVs are likely due to variations in bEV production resulting from strain diversity.https://www.mdpi.com/2076-2607/13/4/753<i>Helicobacter pylori</i> (<i>H. pylori</i>)bacterial extracellular vesicles (bEVs)Lpp20extragastric disease
spellingShingle Aoi Okamoto
Tatsuki Shibuta
Nanaka Morita
Ryota Fujinuma
Masaya Shiraishi
Reimi Matsuda
Mayu Okada
Satoe Watanabe
Tsukuru Umemura
Hiroaki Takeuchi
Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
Microorganisms
<i>Helicobacter pylori</i> (<i>H. pylori</i>)
bacterial extracellular vesicles (bEVs)
Lpp20
extragastric disease
title Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
title_full Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
title_fullStr Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
title_full_unstemmed Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
title_short Identification of Released Bacterial Extracellular Vesicles Containing Lpp20 from <i>Helicobacter pylori</i>
title_sort identification of released bacterial extracellular vesicles containing lpp20 from i helicobacter pylori i
topic <i>Helicobacter pylori</i> (<i>H. pylori</i>)
bacterial extracellular vesicles (bEVs)
Lpp20
extragastric disease
url https://www.mdpi.com/2076-2607/13/4/753
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