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 journalJournal articleResearchpeer-review

4 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 languageEnglish
JournalJournal of Immunological Methods
Volume422
Pages (from-to)59-71
ISSN0022-1759
DOIs
Publication statusPublished - 2015

Fingerprint

Interleukin-1
Lipopolysaccharides
Monocytes
Macrophages
Monoclonal Antibodies
Peptides
Calpain
Fluorescein
Cell Nucleus
Flow Cytometry
Peptide Hydrolases
Cytokines
Inflammation

Keywords

  • Interleukin-1 alpha

Cite this

Carlsen, Thomas Gelsing ; Kjærsgaard, Pernille ; Jørgensen, Trine Lykke ; Foldbjerg, Rasmus ; Nielsen, Mads Lausen ; Guldbæk Poulsen, Thomas Bouet ; Zabieglo, Katarzyna ; Christiansen, Gunna ; Birkelund, Svend. / Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages. In: Journal of Immunological Methods. 2015 ; Vol. 422. pp. 59-71.
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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",
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Interleukin-1α activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages. / Carlsen, Thomas Gelsing; Kjærsgaard, Pernille; Jørgensen, Trine Lykke; Foldbjerg, Rasmus; Nielsen, Mads Lausen; Guldbæk Poulsen, Thomas Bouet; Zabieglo, Katarzyna; Christiansen, Gunna; Birkelund, Svend.

In: Journal of Immunological Methods, Vol. 422, 2015, p. 59-71.

Research output: Contribution to journalJournal articleResearchpeer-review

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

VL - 422

SP - 59

EP - 71

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -