Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome
Osteopontin (OPN) mediates bone remodeling and tissue debridement. The OPN protein is cleaved, but it is unclear how full-length (FL)-OPN or its cleaved form perform their biological activities in target cells. We, therefore, performed the molecular characterization of OPN in exosomes (Exo). The Exo...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696453/ https://www.ncbi.nlm.nih.gov/pubmed/33187327 http://dx.doi.org/10.3390/ijms21228490 |
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author | Bai, Gaowa Matsuba, Takashi Niki, Toshiro Hattori, Toshio |
author_facet | Bai, Gaowa Matsuba, Takashi Niki, Toshiro Hattori, Toshio |
author_sort | Bai, Gaowa |
collection | PubMed |
description | Osteopontin (OPN) mediates bone remodeling and tissue debridement. The OPN protein is cleaved, but it is unclear how full-length (FL)-OPN or its cleaved form perform their biological activities in target cells. We, therefore, performed the molecular characterization of OPN in exosomes (Exo). The Exo were isolated from lipopolysaccharide (LPS)-stimulated phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages. The Exo were also isolated from PMA-differentiated THP-1 macrophages. The Exo were identified using the qNano multiple analyzer (diameter 59–315 nm) and western blotting with a CD9 antibody. LPS-stimulated cells produced more particles than non-stimulated cells. The presence of the FL or the cleaved form of OPN was confirmed using western blot analysis. A mixture of FL and cleaved OPN was also measured using an ELISA system (Ud-OPN) and their presence in the Exo was confirmed. Ud/FL ratios became low after LPS stimulation, indicating the enhanced encapsulation of FL-OPN in the Exo by LPS. These findings suggest that LPS stimulation of human macrophages facilitates the synthesis of FL-OPN, which is cleaved in cells or the Exo after release. These findings indicate that Exo is a suitable vehicle to transfer OPN to the target cells. |
format | Online Article Text |
id | pubmed-7696453 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76964532020-11-29 Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome Bai, Gaowa Matsuba, Takashi Niki, Toshiro Hattori, Toshio Int J Mol Sci Article Osteopontin (OPN) mediates bone remodeling and tissue debridement. The OPN protein is cleaved, but it is unclear how full-length (FL)-OPN or its cleaved form perform their biological activities in target cells. We, therefore, performed the molecular characterization of OPN in exosomes (Exo). The Exo were isolated from lipopolysaccharide (LPS)-stimulated phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages. The Exo were also isolated from PMA-differentiated THP-1 macrophages. The Exo were identified using the qNano multiple analyzer (diameter 59–315 nm) and western blotting with a CD9 antibody. LPS-stimulated cells produced more particles than non-stimulated cells. The presence of the FL or the cleaved form of OPN was confirmed using western blot analysis. A mixture of FL and cleaved OPN was also measured using an ELISA system (Ud-OPN) and their presence in the Exo was confirmed. Ud/FL ratios became low after LPS stimulation, indicating the enhanced encapsulation of FL-OPN in the Exo by LPS. These findings suggest that LPS stimulation of human macrophages facilitates the synthesis of FL-OPN, which is cleaved in cells or the Exo after release. These findings indicate that Exo is a suitable vehicle to transfer OPN to the target cells. MDPI 2020-11-11 /pmc/articles/PMC7696453/ /pubmed/33187327 http://dx.doi.org/10.3390/ijms21228490 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bai, Gaowa Matsuba, Takashi Niki, Toshiro Hattori, Toshio Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title | Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title_full | Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title_fullStr | Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title_full_unstemmed | Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title_short | Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome |
title_sort | stimulation of thp-1 macrophages with lps increased the production of osteopontin-encapsulating exosome |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696453/ https://www.ncbi.nlm.nih.gov/pubmed/33187327 http://dx.doi.org/10.3390/ijms21228490 |
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