Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages

Thomas Gelsing Carlsen; Pernille Kjærsgaard; Trine Lykke Jørgensen; Rasmus Foldbjerg; Mads Lausen Nielsen; Thomas Bouet Guldbæk Poulsen; Katarzyna Zabieglo; Gunna Christiansen; Svend Birkelund

doi:10.1016/j.jim.2015.03.025

Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages

Thomas Gelsing Carlsen, Pernille Kjærsgaard, Trine Lykke Jørgensen, Rasmus Foldbjerg, Mads Lausen Nielsen, Thomas Bouet Guldbæk Poulsen, Katarzyna Zabieglo, Gunna Christiansen, Svend Birkelund

Research output: Contribution to journal › Journal article › Research › peer-review

6 Citations (Scopus)

Abstract

Background: Interleukin-1α (IL-1α) is a proinflammatory cytokine belonging to the IL-1 family. It is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a 16 kDa propiece and a 17 kDa mature IL-1α peptide. In contrast to its close relative, IL-1β, the role of IL- 1α in inflammation is only partly understood.

Results: Human macrophages/monocytes, stimulated with lipopolysaccharide (LPS) were analyzed for production and localization of IL-1α by use of a monoclonal antibody (MAb) generated against IL-1α pro piece. We found that IL-1α propiece was detected within the nuclei of the cells 2 hours (hrs) after LPS stimulation and production continued for up to 20 hrs. The MAb was conjugated to fluorescein isothiocyanate (FITC) for use in flow cytometry. Based on the flow cytometric analysis CD68 positive cells were positive for IL-1α in agreement with CD68 being a marker for monocytes.

Conclusions: Here, we demonstrate, for the first time, a method to visualize and measure the production of IL-1α in both human monocytes and macrophages.

Original language	English
Journal	Journal of Immunological Methods
Volume	422
Pages (from-to)	59-71
ISSN	0022-1759
DOIs	https://doi.org/10.1016/j.jim.2015.03.025
Publication status	Published - 2015

Keywords

Interleukin-1 alpha

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10.1016/j.jim.2015.03.025

http://www.sciencedirect.com/science/article/pii/S0022175915001179

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Interleukin-1 in the pathogenesis and treatment of inflammatory diseases
Carlsen, T. G. & Birkelund, S.
10/04/2012 → 01/12/2014
Project: Research

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@article{a4d2285d277a421099a821e4f9019f36,

title = "Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages",

abstract = "Background: Interleukin-1α (IL-1α) is a proinflammatory cytokine belonging to the IL-1 family. It is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a 16 kDa propiece and a 17 kDa mature IL-1α peptide. In contrast to its close relative, IL-1β, the role of IL- 1α in inflammation is only partly understood.Results: Human macrophages/monocytes, stimulated with lipopolysaccharide (LPS) were analyzed for production and localization of IL-1α by use of a monoclonal antibody (MAb) generated against IL-1α pro piece. We found that IL-1α propiece was detected within the nuclei of the cells 2 hours (hrs) after LPS stimulation and production continued for up to 20 hrs. The MAb was conjugated to fluorescein isothiocyanate (FITC) for use in flow cytometry. Based on the flow cytometric analysis CD68 positive cells were positive for IL-1α in agreement with CD68 being a marker for monocytes. Conclusions: Here, we demonstrate, for the first time, a method to visualize and measure the production of IL-1α in both human monocytes and macrophages.",

keywords = "Interleukin-1 alpha",

author = "Carlsen, {Thomas Gelsing} and Pernille Kj{\ae}rsgaard and J{\o}rgensen, {Trine Lykke} and Rasmus Foldbjerg and Nielsen, {Mads Lausen} and {Guldb{\ae}k Poulsen}, {Thomas Bouet} and Katarzyna Zabieglo and Gunna Christiansen and Svend Birkelund",

year = "2015",

doi = "10.1016/j.jim.2015.03.025",

language = "English",

volume = "422",

pages = "59--71",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

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TY - JOUR

T1 - Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages

AU - Carlsen, Thomas Gelsing

AU - Kjærsgaard, Pernille

AU - Jørgensen, Trine Lykke

AU - Foldbjerg, Rasmus

AU - Nielsen, Mads Lausen

AU - Guldbæk Poulsen, Thomas Bouet

AU - Zabieglo, Katarzyna

AU - Christiansen, Gunna

AU - Birkelund, Svend

PY - 2015

Y1 - 2015

N2 - Background: Interleukin-1α (IL-1α) is a proinflammatory cytokine belonging to the IL-1 family. It is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a 16 kDa propiece and a 17 kDa mature IL-1α peptide. In contrast to its close relative, IL-1β, the role of IL- 1α in inflammation is only partly understood.Results: Human macrophages/monocytes, stimulated with lipopolysaccharide (LPS) were analyzed for production and localization of IL-1α by use of a monoclonal antibody (MAb) generated against IL-1α pro piece. We found that IL-1α propiece was detected within the nuclei of the cells 2 hours (hrs) after LPS stimulation and production continued for up to 20 hrs. The MAb was conjugated to fluorescein isothiocyanate (FITC) for use in flow cytometry. Based on the flow cytometric analysis CD68 positive cells were positive for IL-1α in agreement with CD68 being a marker for monocytes. Conclusions: Here, we demonstrate, for the first time, a method to visualize and measure the production of IL-1α in both human monocytes and macrophages.

AB - Background: Interleukin-1α (IL-1α) is a proinflammatory cytokine belonging to the IL-1 family. It is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a 16 kDa propiece and a 17 kDa mature IL-1α peptide. In contrast to its close relative, IL-1β, the role of IL- 1α in inflammation is only partly understood.Results: Human macrophages/monocytes, stimulated with lipopolysaccharide (LPS) were analyzed for production and localization of IL-1α by use of a monoclonal antibody (MAb) generated against IL-1α pro piece. We found that IL-1α propiece was detected within the nuclei of the cells 2 hours (hrs) after LPS stimulation and production continued for up to 20 hrs. The MAb was conjugated to fluorescein isothiocyanate (FITC) for use in flow cytometry. Based on the flow cytometric analysis CD68 positive cells were positive for IL-1α in agreement with CD68 being a marker for monocytes. Conclusions: Here, we demonstrate, for the first time, a method to visualize and measure the production of IL-1α in both human monocytes and macrophages.

KW - Interleukin-1 alpha

U2 - 10.1016/j.jim.2015.03.025

DO - 10.1016/j.jim.2015.03.025

M3 - Journal article

SN - 0022-1759

VL - 422

SP - 59

EP - 71

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

ER -

Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages

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Keywords

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Interleukin-1 in the pathogenesis and treatment of inflammatory diseases

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