2. Academic Validation
  3. Human TNF-Luc reporter mouse: A new model to quantify inflammatory responses

Human TNF-Luc reporter mouse: A new model to quantify inflammatory responses

  • Sci Rep. 2019 Jan 17;9(1):193. doi: 10.1038/s41598-018-36969-x.
Faisal Minshawi 1 2,  Mike R H White 1,  Werner Muller 1,  Neil Humphreys 1,  Dean Jackson 1,  Barry J Campbell 3,  Antony Adamson 4,  Stamatia Papoutsopoulou 5 6
Affiliations

Affiliations

  • 1 Faculty of Biology, Medicine and Health, School of Biological Sciences, Manchester, M13 9PT, United Kingdom.
  • 2 Department of Laboratory Medicine, Faculty of Applied Medical Science, Umm Al-Qura University, Makkah, Saudi Arabia.
  • 3 Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, United Kingdom.
  • 4 Faculty of Biology, Medicine and Health, School of Biological Sciences, Manchester, M13 9PT, United Kingdom. antony.adamson@manchester.ac.uk.
  • 5 Faculty of Biology, Medicine and Health, School of Biological Sciences, Manchester, M13 9PT, United Kingdom. s.papoutsopoulou@liverpool.ac.uk.
  • 6 Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, United Kingdom. s.papoutsopoulou@liverpool.ac.uk.
PMID: 30655563 DOI: 10.1038/s41598-018-36969-x
Abstract

Tumour necrosis factor (TNF) is a key cytokine during inflammatory responses and its dysregulation is detrimental in many inflammatory diseases, such as rheumatoid arthritis and inflammatory bowel disease. Here, we used a Bacterial artificial chromosome (BAC) construct that expresses luciferase under the control of the human TNF locus to generate a novel transgenic mouse, the hTNF.LucBAC strain. In vitro stimulation of hTNF.LucBAC cells of different origin revealed a cell specific response to stimuli demonstrating the integrated construct's ability as a proxy for inflammatory gene response. Lipopolysaccharide was the most potent luciferase inducer in macrophages, while TNF was a strong activator in intestinal organoids. Lipopolysaccharide-induced luciferase activity in macrophages was downregulated by inhibitors of NF-κB pathway, as well as by Interleukin-10, a known anti-inflammatory cytokine. Moreover, the transgene-dependent luciferase activity showed a positive correlation to the endogenous murine soluble TNF secreted to the culture medium. In conclusion, the hTNF.LucBAC strain is a valuable tool for studying and screening molecules that target TNF synthesis and will allow further functional studies of the regulatory elements of the TNF locus.

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