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 33kDa 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 monocyte derived macrophages, stimulated with lipopolysaccharide (LPS) were analysed for production and localization of IL-1α by use of a monoclonal antibody (MAb) generated against recombinant precursor IL-1α. We found that the MAb detected IL-1α within the nuclei of the cells 2h (hours) after LPS stimulation and production continued for up to 20 h. At no time could we demonstrate cleavage of the IL-1α precursor. 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.
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: tc@hst.aau.dk.
authors: Thomas Gelsing
authors: TG
authors: Carlsen
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: pkpe09@student.aau.dk.
authors: Pernille
authors: P
authors: Kjærsgaard
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: tlj@hst.aau.dk.
authors: Trine Lykke
authors: TL
authors: Jørgensen
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: rfol@hst.aau.dk.
authors: Rasmus
authors: R
authors: Foldbjerg
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: madsni11@student.aau.dk.
authors: Mads Lausen
authors: ML
authors: Nielsen
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark. Electronic address: tbgp11@student.aau.dk.
authors: Thomas Bouet Guldbæk
authors: TB
authors: Poulsen
authors: Loke Diagnostics, Sindalsvej 17, 8240 Risskov, Denmark. Electronic address: katarzyna.zabieglo@uj.edu.pl.
authors: Katarzyna
authors: K
authors: Zabieglo
authors: Loke Diagnostics, Sindalsvej 17, 8240 Risskov, Denmark. Electronic address: gunna@loke.dk.
authors: Gunna
authors: G
authors: Christiansen
authors: Department of Health Science and Technology, Fredrik Bajers Vej 3b, 9220 Aalborg Ø, Denmark; Loke Diagnostics, Sindalsvej 17, 8240 Risskov, Denmark. Electronic address: sbirkelund@hst.aau.dk.
authors: Svend
authors: S
authors: Birkelund
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D001483:Base Sequence; D045744:Cell Line, Tumor; D004789:Enzyme Activation; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D007249:Inflammation / Q000276:immunology*; D053582:Interleukin-1alpha / Q000097:blood / Q000235:genetics / Q000276:immunology*; D008070:Lipopolysaccharides; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D009000:Monocytes / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D017423:Sequence Analysis, RNA; D000097382:CD68 Molecule
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D005456:Fluorescent Dyes; D053582:Interleukin-1alpha; D008070:Lipopolysaccharides; D012333:RNA, Messenger; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C508584:IL1A protein, human
keywords: Flow cytometry; IL-1α; Intranuclear staining; Monoclonal antibody; Monocytes