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Bioactivity and secretion of interleukin-18 (IL-18) generated by equine and feline IL-18 expression constructs.
Abstract: Interleukin 18 (IL-18) is a cytokine capable of induction of IFNgamma, granulocyte monocyte-colony stimulating factor (GM-CSF), TNFalpha and IL-1 in immunocompetent cells. Equine and feline plasmid vectors expressing pro-IL-18, mature IL-18 and IL-18 fused to a synthetic signal sequence from human IL-1beta receptor antagonist protein (ILRAP), ILRAP-IL-18, have been generated. In vitro protein expression of these constructs was compared by Western blot analysis. These data demonstrated that ILRAP-IL-18 protein was secreted readily from transfected chinese hamster ovary (CHO) cells. A simple bioassay for human IL-18 was recently described using human myelomonocytic KG-1 cells, which produce human IFNgamma in response to human IL-18 in a dose dependent manner (Konishi et al., 1997). We demonstrated bioactivity of equine and feline IL-18 protein in transfection products of CHO cells using this assay. Bioactivity of ILRAP-IL-18 protein was demonstrated in the culture medium of transfected CHO cells. These data imply that the ILRAP-IL-18 construct shows potential for use in vivo, where cell secretion of protein is crucial.
Publication Date: 2004-11-16 PubMed ID: 15541795DOI: 10.1016/j.vetimm.2004.08.003Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This article explores the production and potential utility of Interleukin 18 (IL-18), a cytokine that influences immune responses, generated by encoded equine and feline vectors.
Understanding Interleukin 18 (IL-18)
- Interleukin 18 (IL-18) is a cytokine (a type of immunological messenger molecule) that is capable of inducing the production other important substances within immune cells, such as interferon-gamma (IFNgamma), granulocyte monocyte-colony stimulating factor (GM-CSF), tumor necrosis factor alpha (TNFalpha), and Interleukin-1 (IL-1).
- The cytokines play a crucial role in coordinating immune responses in the body, including inflammation and the defense against pathogens.
Construction of IL-18 Encoding Vectors
- The study reports the creation of equine and feline plasmid vectors that are able to express pro-IL-18 (an inactive precursor), mature IL-18, and a synthetic fusion of IL-18 with a signal sequence taken from human Interleukin-1 beta receptor antagonist protein (ILRAP), named ILRAP-IL-18.
- The use of the IL-18 encoding vectors is a molecular biology technique used to multiply the amounts of IL-18 in cells for therapeutic purposes.
In-vitro Protein Expression
- Analysis carried out via Western Blot (a laboratory method used for detection of specific protein molecules from complex mixtures) showcased a comparison of the protein expression capabilities of the different IL-18 constructs.
- The data demonstrated that the ILRAP-IL-18 protein could be effectively secreted from transfected Chinese Hamster Ovary (CHO) cells, a type of cell commonly used in biological and medical research.
Bioactivity of Constructs
- The study also revealed that both the equine and feline IL-18 proteins showed bioactivity (the ability to have a biological effect) post transfection into the CHO cells.
- A previously described bioassay method involving human myelomonocytic KG-1 cells was used to demonstrate this. The KG-1 cells produce interferon gamma in response to human IL-18 in a dose-dependent manner, indicating the bioactivity of IL-18.
- The fused ILRAP-IL-18 protein also demonstrated bioactivity in the culture medium of the transfected CHO cells, implying its potential for use in vivo where secretion of the protein from cells is crucial.
In conclusion, the ILRAP-IL-18 construct shows promising potential for biological activity in vivo, underlining its potential therapeutic utility.
Cite This Article
APA
O'Donovan LH, McMonagle EL, Taylor S, Argyle DJ, Nicolson L.
(2004).
Bioactivity and secretion of interleukin-18 (IL-18) generated by equine and feline IL-18 expression constructs.
Vet Immunol Immunopathol, 102(4), 421-428.
https://doi.org/10.1016/j.vetimm.2004.08.003 Publication
Researcher Affiliations
- Division of Infection and Immunity, Institute of Comparative Medicine, University of Glasgow, 464 Bearsden Road, Garscube Estate, Glasgow G61 1QH, UK. l.o'donovan@bio.gla.ac.uk
MeSH Terms
- Animals
- Biological Assay
- CHO Cells
- Cats / immunology
- Cricetinae
- Cricetulus
- Gene Expression Regulation / immunology
- Horses / immunology
- In Vitro Techniques
- Interferon-gamma / biosynthesis
- Interleukin 1 Receptor Antagonist Protein
- Interleukin-18 / analysis
- Interleukin-18 / biosynthesis
- Recombinant Fusion Proteins / biosynthesis
- Sialoglycoproteins / genetics
Citations
This article has been cited 2 times.- Liu Y, Sato H, Hamana M, Moonan NA, Yoneda M, Xia X, Kai C. Construction of an expression system for bioactive IL-18 and generation of recombinant canine distemper virus expressing IL-18. J Vet Med Sci 2014 Sep;76(9):1241-8.
- O'Donovan LH, McMonagle EL, Taylor S, Bain D, Pacitti AM, Golder MC, McDonald M, Hanlon L, Onions DE, Argyle DJ, Jarrett O, Nicolson L. A vector expressing feline mature IL-18 fused to IL-1beta antagonist protein signal sequence is an effective adjuvant to a DNA vaccine for feline leukaemia virus. Vaccine 2005 May 31;23(29):3814-23.
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