An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test

Abstract Endotoxin neutralization, caused by plasma components, makes it difficult to detect endotoxins in human blood. In this study, we investigated which factors influence the recovery of endotoxins using limulus ameobocyte lysate (LAL)-based assays. The individual factors that were examined in m...

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Autores principales:	Stephan Harm, Claudia Schildböck, Karin Strobl, Jens Hartmann
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/57bb840215054b33bc349753284342dd
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spelling	oai:doaj.org-article:57bb840215054b33bc349753284342dd2021-12-02T14:21:59ZAn in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test10.1038/s41598-021-83487-42045-2322https://doaj.org/article/57bb840215054b33bc349753284342dd2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83487-4https://doaj.org/toc/2045-2322Abstract Endotoxin neutralization, caused by plasma components, makes it difficult to detect endotoxins in human blood. In this study, we investigated which factors influence the recovery of endotoxins using limulus ameobocyte lysate (LAL)-based assays. The individual factors that were examined in more detail were lipoprotein content, type of blood anticoagulation, kinetics and serum levels of divalent cations. Furthermore, it was investigated whether there is a direct correlation between LAL activity and monocyte activation. We could show that polyanionic heparin increases endotoxin recovery in blood, while citrate anticoagulation promotes endotoxin neutralization. Furthermore, we could show that the endotoxin activity in human plasma and serum decreases strongly over time. Time-dependent endotoxin neutralization reaches its maximum after 4–6 h incubation. By means of filtration tests we could determine that endotoxins in the plasma bind to lipoproteins but do not influence their activity. Comparative measurements have shown that high LAL activity of endotoxins in plasma simultaneously possesses high monocyte activating properties in whole blood. For the maximum recovery of endotoxins in human blood the physiological calcium and magnesium concentrations are sufficient. In this study, it was shown that the endotoxin neutralizing plasma components have a molecular weight similar to β2-microglobulin (11.7 kDa). For the exact identification of the endotoxin neutralizing plasma components, which caused a modulation of the immunostimulating endotoxin activity, further investigations have to be carried out in the future.Stephan HarmClaudia SchildböckKarin StroblJens HartmannNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-23 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Stephan Harm Claudia Schildböck Karin Strobl Jens Hartmann An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
description	Abstract Endotoxin neutralization, caused by plasma components, makes it difficult to detect endotoxins in human blood. In this study, we investigated which factors influence the recovery of endotoxins using limulus ameobocyte lysate (LAL)-based assays. The individual factors that were examined in more detail were lipoprotein content, type of blood anticoagulation, kinetics and serum levels of divalent cations. Furthermore, it was investigated whether there is a direct correlation between LAL activity and monocyte activation. We could show that polyanionic heparin increases endotoxin recovery in blood, while citrate anticoagulation promotes endotoxin neutralization. Furthermore, we could show that the endotoxin activity in human plasma and serum decreases strongly over time. Time-dependent endotoxin neutralization reaches its maximum after 4–6 h incubation. By means of filtration tests we could determine that endotoxins in the plasma bind to lipoproteins but do not influence their activity. Comparative measurements have shown that high LAL activity of endotoxins in plasma simultaneously possesses high monocyte activating properties in whole blood. For the maximum recovery of endotoxins in human blood the physiological calcium and magnesium concentrations are sufficient. In this study, it was shown that the endotoxin neutralizing plasma components have a molecular weight similar to β2-microglobulin (11.7 kDa). For the exact identification of the endotoxin neutralizing plasma components, which caused a modulation of the immunostimulating endotoxin activity, further investigations have to be carried out in the future.
format	article
author	Stephan Harm Claudia Schildböck Karin Strobl Jens Hartmann
author_facet	Stephan Harm Claudia Schildböck Karin Strobl Jens Hartmann
author_sort	Stephan Harm
title	An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
title_short	An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
title_full	An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
title_fullStr	An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
title_full_unstemmed	An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test
title_sort	in vitro study on factors affecting endotoxin neutralization in human plasma using the limulus amebocyte lysate test
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/57bb840215054b33bc349753284342dd
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An in vitro study on factors affecting endotoxin neutralization in human plasma using the Limulus amebocyte lysate test

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