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Ficolin-1 is present in a highly mobilizable subset of human neutrophil granules and associates with the cell surface after stimulation with fMLP

Rørvig, Sara ; Honore, Christian ; Larsson, Lars-Inge ; Ohlsson, Sophie LU orcid ; Pedersen, Corinna C ; Jacobsen, Lars C ; Cowland, Jack B ; Garred, Peter and Borregaard, Niels (2009) In Journal of Leukocyte Biology 86(6). p.49-1439
Abstract

Ficolins are soluble molecules that bind carbohydrate present on the surface of microorganisms and function as recognition molecules in the lectin complement pathway. Three ficolins have been identified in humans: ficolin-1, ficolin-2, and ficolin-3. Ficolin-1 is synthesized in monocytes and type II alveolar epithelial cells. Ficolin-1 has been shown to be present in secretory granules of human neutrophils, but it is not known which subset of the neutrophils' secretory granules harbors ficolin-1. To determine the exact subcellular localization of ficolin-1 in neutrophils, recombinant ficolin-1 was expressed in Chinese hamster ovary cells and used for generation of polyclonal antibodies. This allowed detection of ficolin-1 in subcellular... (More)

Ficolins are soluble molecules that bind carbohydrate present on the surface of microorganisms and function as recognition molecules in the lectin complement pathway. Three ficolins have been identified in humans: ficolin-1, ficolin-2, and ficolin-3. Ficolin-1 is synthesized in monocytes and type II alveolar epithelial cells. Ficolin-1 has been shown to be present in secretory granules of human neutrophils, but it is not known which subset of the neutrophils' secretory granules harbors ficolin-1. To determine the exact subcellular localization of ficolin-1 in neutrophils, recombinant ficolin-1 was expressed in Chinese hamster ovary cells and used for generation of polyclonal antibodies. This allowed detection of ficolin-1 in subcellular fractions of human neutrophils by ELISA, by Western blotting, and by immunohistochemistry. Real-time PCR examination of normal human bone marrow showed FCN1 gene expression largely in myelocytes, metamyelocytes, and band cells with a profile quite similar to that of gelatinase. In accordance with this, biosynthesis studies of neutrophils precursor cells showed that ficolin-1 was primarily synthesized in myelocytes, metamyelocytes, and band cells. Immunohistochemistry and subcellular fractionation demonstrated that ficolin-1 is primarily localized in gelatinase granules but also in highly exocytosable gelatinase-poor granules, not described previously. Ficolin-1 is released from neutrophil granules by stimulation with fMLP or PMA, and the majority becomes associated with the surface membrane of the cells and can be detected by flow cytometry. Our studies show that neutrophils are a major source of ficolin-1, which can be readily exocytosed by stimulation.

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author
; ; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, CHO Cells, Carcinogens/pharmacology, Cricetinae, Cricetulus, Exocytosis/drug effects, Gene Expression Regulation/drug effects, Humans, Lectins/immunology, N-Formylmethionine Leucyl-Phenylalanine/pharmacology, Neutrophils/immunology, Recombinant Proteins/immunology, Secretory Vesicles/immunology, Tetradecanoylphorbol Acetate/pharmacology
in
Journal of Leukocyte Biology
volume
86
issue
6
pages
11 pages
publisher
John Wiley & Sons Inc.
external identifiers
  • pmid:19741154
  • scopus:73149096727
ISSN
1938-3673
DOI
10.1189/jlb.1008606
language
English
LU publication?
no
id
c6d00b5d-cf13-427d-a0a9-58dff5c3d870
date added to LUP
2018-06-16 22:31:24
date last changed
2024-04-01 07:19:32
@article{c6d00b5d-cf13-427d-a0a9-58dff5c3d870,
  abstract     = {{<p>Ficolins are soluble molecules that bind carbohydrate present on the surface of microorganisms and function as recognition molecules in the lectin complement pathway. Three ficolins have been identified in humans: ficolin-1, ficolin-2, and ficolin-3. Ficolin-1 is synthesized in monocytes and type II alveolar epithelial cells. Ficolin-1 has been shown to be present in secretory granules of human neutrophils, but it is not known which subset of the neutrophils' secretory granules harbors ficolin-1. To determine the exact subcellular localization of ficolin-1 in neutrophils, recombinant ficolin-1 was expressed in Chinese hamster ovary cells and used for generation of polyclonal antibodies. This allowed detection of ficolin-1 in subcellular fractions of human neutrophils by ELISA, by Western blotting, and by immunohistochemistry. Real-time PCR examination of normal human bone marrow showed FCN1 gene expression largely in myelocytes, metamyelocytes, and band cells with a profile quite similar to that of gelatinase. In accordance with this, biosynthesis studies of neutrophils precursor cells showed that ficolin-1 was primarily synthesized in myelocytes, metamyelocytes, and band cells. Immunohistochemistry and subcellular fractionation demonstrated that ficolin-1 is primarily localized in gelatinase granules but also in highly exocytosable gelatinase-poor granules, not described previously. Ficolin-1 is released from neutrophil granules by stimulation with fMLP or PMA, and the majority becomes associated with the surface membrane of the cells and can be detected by flow cytometry. Our studies show that neutrophils are a major source of ficolin-1, which can be readily exocytosed by stimulation.</p>}},
  author       = {{Rørvig, Sara and Honore, Christian and Larsson, Lars-Inge and Ohlsson, Sophie and Pedersen, Corinna C and Jacobsen, Lars C and Cowland, Jack B and Garred, Peter and Borregaard, Niels}},
  issn         = {{1938-3673}},
  keywords     = {{Animals; CHO Cells; Carcinogens/pharmacology; Cricetinae; Cricetulus; Exocytosis/drug effects; Gene Expression Regulation/drug effects; Humans; Lectins/immunology; N-Formylmethionine Leucyl-Phenylalanine/pharmacology; Neutrophils/immunology; Recombinant Proteins/immunology; Secretory Vesicles/immunology; Tetradecanoylphorbol Acetate/pharmacology}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{49--1439}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Journal of Leukocyte Biology}},
  title        = {{Ficolin-1 is present in a highly mobilizable subset of human neutrophil granules and associates with the cell surface after stimulation with fMLP}},
  url          = {{http://dx.doi.org/10.1189/jlb.1008606}},
  doi          = {{10.1189/jlb.1008606}},
  volume       = {{86}},
  year         = {{2009}},
}