Toxicol Sci. 2017 Apr 21. doi: 10.1093/toxsci/kfx069. [Epub ahead of print]
Ogese MO1, Faulkner L2, Jenkins RE2, French NS2, Copple IM2, Antoine DJ2, Elmasry M2,3, Malik H3, Goldring CE2, Kevin Park B2, Betts C1, Naisbitt DJ2.
1 Pathology Sciences, Drug Safety and Metabolism, AstraZeneca R&D, Darwin Building 310, Cambridge Science Park, Milton Rd, Cambridge CB4 0WG, United Kingdom.
2 MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, University of Liverpool, Ashton Street, Liverpool L69 3GE, United Kingdom.
3 North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK.
It is now apparent that antigen-specific T-cells are activated in certain patients with drug-induced liver injury (DILI). Since cross-talk between hepatocytes and immune cells is likely to be critical in determining the outcome of drug exposure, the aim of this study was to profile the signals released by drug-treated hepatocytes and to characterise the impact of these molecules on dendritic cells. Human hepatocytes were exposed to three drugs (flucloxacillin, amoxicillin and isoniazid) associated with DILI potentially mediated by the adaptive immune system as drug-specific T-cells have been isolated from DILI patients, and the metabolite nitroso-sulfamethoxazole (SMX-NO). Hepatocyte toxicity, cytokine release and activation of oxidative stress pathways were measured. Supernatants were transferred to monocyte-derived dendritic cells and cell phenotype and function were assessed. High-mobility group box 1 protein (HMGB1) and lactate dehydrogenase release as well as adenosine triphosphate depletion occurred in a drug-, time- and concentration-dependent manner with SMX-NO and flucloxacillin, whereas isoniazid and amoxicillin were non-toxic. Furthermore, drug-induced activation of Nrf2 marker genes was observed when hepatocytes were exposed to test drugs. The disulphide isoform of HMGB1 stimulated dendritic cell cytokine release and enhanced the priming of naive T cells. Incubation of dendritic cells with supernatant from drug-treated hepatocytes resulted in two distinct cytokine profiles. SMX-NO/flucloxacillin stimulated secretion of TNF-á, IL-6, IL-1á and IL-1-â. Isoniazid which did not induce significant hepatocyte toxicity, compared with SMX-NO and flucloxacillin, stimulated the release of a panel of cytokines including the above and IFN-γ, IL-12, IL-17A, IP-10 and IL-10. Collectively, our study identifies drug-specific signalling pathways between hepatocytes and immune cells that could influence whether drug exposure will result in an immune response and tissue injury.
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