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Article

Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome

1
Department of Health Science and Social Welfare, Kibi International University, Takahashi 716-8508, Japan
2
Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8503, Japan
3
Department of Immunology, Kagawa University, Kita-gun, Kagawa 7610793, Japan
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(22), 8490; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228490
Received: 28 October 2020 / Accepted: 9 November 2020 / Published: 11 November 2020
(This article belongs to the Special Issue Extracellular Vesicles in Inflammation)
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. View Full-Text
Keywords: osteopontin; THP-1; exosome; LPS; Exo-Prep osteopontin; THP-1; exosome; LPS; Exo-Prep
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MDPI and ACS Style

Bai, G.; Matsuba, T.; Niki, T.; Hattori, T. Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome. Int. J. Mol. Sci. 2020, 21, 8490. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228490

AMA Style

Bai G, Matsuba T, Niki T, Hattori T. Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome. International Journal of Molecular Sciences. 2020; 21(22):8490. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228490

Chicago/Turabian Style

Bai, Gaowa, Takashi Matsuba, Toshiro Niki, and Toshio Hattori. 2020. "Stimulation of THP-1 Macrophages with LPS Increased the Production of Osteopontin-Encapsulating Exosome" International Journal of Molecular Sciences 21, no. 22: 8490. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228490

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