Human TNF-Luc reporter mouse: A new model to quantify inflammatory responses.
Faisal MinshawiMike R H WhiteWerner MullerNeil HumphreysDean JacksonBarry J CampbellAntony D AdamsonStamatia PapoutsopoulouPublished in: Scientific reports (2019)
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.
Keyphrases
- rheumatoid arthritis
- lipopolysaccharide induced
- endothelial cells
- disease activity
- inflammatory response
- oxidative stress
- signaling pathway
- interstitial lung disease
- induced apoptosis
- ankylosing spondylitis
- copy number
- stem cells
- induced pluripotent stem cells
- lps induced
- nuclear factor
- transcription factor
- immune response
- crispr cas
- toll like receptor
- pluripotent stem cells
- systemic sclerosis
- cell cycle arrest