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Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles
BACKGROUND: Membrane vesicles (MVs) are nanoscale vesicular structures produced by bacteria during their growth in vitro and in vivo. Some bacterial components can be loaded in bacterial MVs, but the roles of the loaded MV molecules are unclear. METHODS: MVs of Staphylococcus aureus RN4220 and its d...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10588482/ https://www.ncbi.nlm.nih.gov/pubmed/37869662 http://dx.doi.org/10.3389/fmicb.2023.1254367 |
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author | Chen, Juan Lv, Yuhuan Shang, Weilong Yang, Yi Wang, Yuting Hu, Zhen Huang, Xiaonan Zhang, Rong Yuan, Jizhen Huang, Jingbin Rao, Xiancai |
author_facet | Chen, Juan Lv, Yuhuan Shang, Weilong Yang, Yi Wang, Yuting Hu, Zhen Huang, Xiaonan Zhang, Rong Yuan, Jizhen Huang, Jingbin Rao, Xiancai |
author_sort | Chen, Juan |
collection | PubMed |
description | BACKGROUND: Membrane vesicles (MVs) are nanoscale vesicular structures produced by bacteria during their growth in vitro and in vivo. Some bacterial components can be loaded in bacterial MVs, but the roles of the loaded MV molecules are unclear. METHODS: MVs of Staphylococcus aureus RN4220 and its derivatives were prepared. Dynamic light scattering analysis was used to evaluate the size distribution, and 4D-label-free liquid chromatography–tandem mass spectrometry analysis was performed to detect protein composition in the MVs. The site-mutation S. aureus RN4220-Δhld and agrA deletion mutant RN4220-ΔagrA were generated via allelic replacement strategies. A hemolysis assay was performed with rabbit red blood cells. CCK-8 and lactate dehydrogenase release assays were used to determine the cytotoxicity of S. aureus MVs against RAW264.7 macrophages. The serum levels of inflammatory factors such as IL-6, IL-1β, and TNFα in mice treated with S. aureus MVs were detected with an enzyme-linked immunosorbent assay kit. RESULTS: Delta-hemolysin (Hld) was identified as a major loaded factor in S. aureus MVs. Further study showed that Hld could promote the production of staphylococcal MVs with smaller sizes. Loaded Hld affected the diversity of loaded proteins in MVs of S. aureus RN4220. Hld resulted in decreased protein diversity in MVs of S. aureus. Site-mutation (RN4220-Δhld) and agrA deletion (RN4220-ΔagrA) mutants produced MVs ((Δhld)MVs and (ΔagrA)MVs) with a greater number of bacterial proteins than those derived from wild-type RN4220 ((wt)MVs). Moreover, Hld contributed to the hemolytic activity of (wt)MVs. Hld-loaded (wt)MVs were cytotoxic to macrophage RAW264.7 cells and could stimulate the production of inflammatory factor IL-6 in vivo. CONCLUSION: This study presented that Hld was a major loaded factor in S. aureus MVs, and the loaded Hld played vital roles in the MV-property modification. |
format | Online Article Text |
id | pubmed-10588482 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-105884822023-10-21 Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles Chen, Juan Lv, Yuhuan Shang, Weilong Yang, Yi Wang, Yuting Hu, Zhen Huang, Xiaonan Zhang, Rong Yuan, Jizhen Huang, Jingbin Rao, Xiancai Front Microbiol Microbiology BACKGROUND: Membrane vesicles (MVs) are nanoscale vesicular structures produced by bacteria during their growth in vitro and in vivo. Some bacterial components can be loaded in bacterial MVs, but the roles of the loaded MV molecules are unclear. METHODS: MVs of Staphylococcus aureus RN4220 and its derivatives were prepared. Dynamic light scattering analysis was used to evaluate the size distribution, and 4D-label-free liquid chromatography–tandem mass spectrometry analysis was performed to detect protein composition in the MVs. The site-mutation S. aureus RN4220-Δhld and agrA deletion mutant RN4220-ΔagrA were generated via allelic replacement strategies. A hemolysis assay was performed with rabbit red blood cells. CCK-8 and lactate dehydrogenase release assays were used to determine the cytotoxicity of S. aureus MVs against RAW264.7 macrophages. The serum levels of inflammatory factors such as IL-6, IL-1β, and TNFα in mice treated with S. aureus MVs were detected with an enzyme-linked immunosorbent assay kit. RESULTS: Delta-hemolysin (Hld) was identified as a major loaded factor in S. aureus MVs. Further study showed that Hld could promote the production of staphylococcal MVs with smaller sizes. Loaded Hld affected the diversity of loaded proteins in MVs of S. aureus RN4220. Hld resulted in decreased protein diversity in MVs of S. aureus. Site-mutation (RN4220-Δhld) and agrA deletion (RN4220-ΔagrA) mutants produced MVs ((Δhld)MVs and (ΔagrA)MVs) with a greater number of bacterial proteins than those derived from wild-type RN4220 ((wt)MVs). Moreover, Hld contributed to the hemolytic activity of (wt)MVs. Hld-loaded (wt)MVs were cytotoxic to macrophage RAW264.7 cells and could stimulate the production of inflammatory factor IL-6 in vivo. CONCLUSION: This study presented that Hld was a major loaded factor in S. aureus MVs, and the loaded Hld played vital roles in the MV-property modification. Frontiers Media S.A. 2023-10-06 /pmc/articles/PMC10588482/ /pubmed/37869662 http://dx.doi.org/10.3389/fmicb.2023.1254367 Text en Copyright © 2023 Chen, Lv, Shang, Yang, Wang, Hu, Huang, Zhang, Yuan, Huang and Rao. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Chen, Juan Lv, Yuhuan Shang, Weilong Yang, Yi Wang, Yuting Hu, Zhen Huang, Xiaonan Zhang, Rong Yuan, Jizhen Huang, Jingbin Rao, Xiancai Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title | Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title_full | Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title_fullStr | Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title_full_unstemmed | Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title_short | Loaded delta-hemolysin shapes the properties of Staphylococcus aureus membrane vesicles |
title_sort | loaded delta-hemolysin shapes the properties of staphylococcus aureus membrane vesicles |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10588482/ https://www.ncbi.nlm.nih.gov/pubmed/37869662 http://dx.doi.org/10.3389/fmicb.2023.1254367 |
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