Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion

The importance of alveolar macrophages has been reported in many toxicology/immunology studies. Alveolar macrophages release interleukin (IL)-1α as a damage-associated molecular pattern (DAMP) when stimulated by fine particles. However, it is unclear whether cell isolation procedures affect ex vivo...

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Autores principales:	Eita Sasaki, Haruka Momose, Keiko Furuhata, Takuo Mizukami, Isao Hamaguchi
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	alveolar macrophages il-1α rip3k necroptosis damage-associated molecular pattern pulmonary toxicity fine particles Immunologic diseases. Allergy RC581-607 Toxicology. Poisons RA1190-1270
Acceso en línea:	https://doaj.org/article/bf20f18bee534a4699ffdafd55d99cb2
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spelling	oai:doaj.org-article:bf20f18bee534a4699ffdafd55d99cb22021-11-17T14:21:57ZImpact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion1547-691X1547-690110.1080/1547691X.2021.1979699https://doaj.org/article/bf20f18bee534a4699ffdafd55d99cb22021-01-01T00:00:00Zhttp://dx.doi.org/10.1080/1547691X.2021.1979699https://doaj.org/toc/1547-691Xhttps://doaj.org/toc/1547-6901The importance of alveolar macrophages has been reported in many toxicology/immunology studies. Alveolar macrophages release interleukin (IL)-1α as a damage-associated molecular pattern (DAMP) when stimulated by fine particles. However, it is unclear whether cell isolation procedures affect ex vivo particle-induced responses in primary mouse alveolar macrophages (mAM). In this study, effects of injection buffer volume used to perform bronchoalveolar lavage fluid (BALF) collection to isolate mAM for use in ex vivo particle-induced responses were assessed. Among the mAM obtained from BALF collected using a 0.55 or 0.75 ml, but not a 1.0 ml buffer injection volume, decreased cell viability and IL-1α release were observed when cells were stimulated ex vivo with silica crystal or aluminum salt. Injected buffer composition did not affect the IL-1α release. On the other hand, IL-6 secretion induced by lipopolysaccharide (LPS) did not differ among mAM obtained from BALF collected using the different volumes. Expression levels of cell surface markers like CD11c, SiglecF, and CD64 did not differ among mAM obtained from BALF collected using the different injection buffer volumes. IL-1α release (and also necroptosis) induced by ex vivoparticle stimulation was suppressed by RIPK3 inhibitor or cytochalasin D co-treatment. Decreases in RIPK3 phosphorylation were noted in mAM obtained in BALF collected using the 1.0 ml injection volume compared with mAM obtained in BALF using 0.55 or 0.75 ml buffer. These observations illustrate that larger volumes of buffer used to collect BALF from mice can affect sensitivity of the isolated mAM to ex vivo particle-induced responses by inhibiting their functions.Eita SasakiHaruka MomoseKeiko FuruhataTakuo MizukamiIsao HamaguchiTaylor & Francis Grouparticlealveolar macrophagesil-1αrip3knecroptosisdamage-associated molecular patternpulmonary toxicityfine particlesImmunologic diseases. AllergyRC581-607Toxicology. PoisonsRA1190-1270ENJournal of Immunotoxicology, Vol 18, Iss 1, Pp 163-172 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	alveolar macrophages il-1α rip3k necroptosis damage-associated molecular pattern pulmonary toxicity fine particles Immunologic diseases. Allergy RC581-607 Toxicology. Poisons RA1190-1270
spellingShingle	alveolar macrophages il-1α rip3k necroptosis damage-associated molecular pattern pulmonary toxicity fine particles Immunologic diseases. Allergy RC581-607 Toxicology. Poisons RA1190-1270 Eita Sasaki Haruka Momose Keiko Furuhata Takuo Mizukami Isao Hamaguchi Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
description	The importance of alveolar macrophages has been reported in many toxicology/immunology studies. Alveolar macrophages release interleukin (IL)-1α as a damage-associated molecular pattern (DAMP) when stimulated by fine particles. However, it is unclear whether cell isolation procedures affect ex vivo particle-induced responses in primary mouse alveolar macrophages (mAM). In this study, effects of injection buffer volume used to perform bronchoalveolar lavage fluid (BALF) collection to isolate mAM for use in ex vivo particle-induced responses were assessed. Among the mAM obtained from BALF collected using a 0.55 or 0.75 ml, but not a 1.0 ml buffer injection volume, decreased cell viability and IL-1α release were observed when cells were stimulated ex vivo with silica crystal or aluminum salt. Injected buffer composition did not affect the IL-1α release. On the other hand, IL-6 secretion induced by lipopolysaccharide (LPS) did not differ among mAM obtained from BALF collected using the different volumes. Expression levels of cell surface markers like CD11c, SiglecF, and CD64 did not differ among mAM obtained from BALF collected using the different injection buffer volumes. IL-1α release (and also necroptosis) induced by ex vivoparticle stimulation was suppressed by RIPK3 inhibitor or cytochalasin D co-treatment. Decreases in RIPK3 phosphorylation were noted in mAM obtained in BALF collected using the 1.0 ml injection volume compared with mAM obtained in BALF using 0.55 or 0.75 ml buffer. These observations illustrate that larger volumes of buffer used to collect BALF from mice can affect sensitivity of the isolated mAM to ex vivo particle-induced responses by inhibiting their functions.
format	article
author	Eita Sasaki Haruka Momose Keiko Furuhata Takuo Mizukami Isao Hamaguchi
author_facet	Eita Sasaki Haruka Momose Keiko Furuhata Takuo Mizukami Isao Hamaguchi
author_sort	Eita Sasaki
title	Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
title_short	Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
title_full	Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
title_fullStr	Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
title_full_unstemmed	Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion
title_sort	impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced il-1α secretion
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/bf20f18bee534a4699ffdafd55d99cb2
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_version_	1718425507182149632

Impact of injection buffer volume to perform bronchoalveolar lavage fluid collection for isolating alveolar macrophages to investigate fine particle-induced IL-1α secretion

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