Subtoxic Concentrations of Hepatotoxic Drugs Lead to Kupffer Cell Activation in a Human In Vitro Liver Model: An Approach to Study DILI.
Drug induced liver injury (DILI) is an idiosyncratic adverse drug reaction leading to severe liver damage. Kupffer cells (KC) sense hepatic tissue stress/damage and therefore could be a tool for the estimation of consequent effects associated with DILI. Aim of the present study was to establish a human in vitro liver model for the investigation of immune-mediated signaling in the pathogenesis of DILI. Hepatocytes and KC were isolated from human liver specimens. The isolated KC yield was 1.2 ± 0.9 × 10(6) cells/g liver tissue with a purity of >80%. KC activation was investigated by the measurement of reactive oxygen intermediates (ROI, DCF assay) and cell activity (XTT assay). The initial KC activation levels showed broad donor variability. Additional activation of KC using supernatants of hepatocytes treated with hepatotoxic drugs increased KC activity and led to donor-dependent changes in the formation of ROI compared to KC incubated with supernatants from untreated hepatocytes. Additionally, a compound- and donor-dependent increase in proinflammatory cytokines or in anti-inflammatory cytokines was detected. In conclusion, KC related immune signaling in hepatotoxicity was successfully determined in a newly established in vitro liver model. KC were able to detect hepatocyte stress/damage and to transmit a donor- and compound-dependent immune response via cytokine production.
Department of General, Visceral and Transplantation Surgery, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Victoria
V
Kegel
Department of General, Visceral and Transplantation Surgery, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Elisa
E
Pfeiffer
BG Trauma Center, Siegfried Weller Institute, Eberhard Karls University Tübingen, Schnarrenbergstrasse 95, 72076 Tübingen, Germany.
Britta
B
Burkhardt
Department of General, Visceral and Transplantation Surgery, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Jia L
JL
Liu
Bioreactor Group, Berlin-Brandenburg Centre for Regenerative Therapies (BCRT), Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Katrin
K
Zeilinger
BG Trauma Center, Siegfried Weller Institute, Eberhard Karls University Tübingen, Schnarrenbergstrasse 95, 72076 Tübingen, Germany.
Andreas K
AK
Nüssler
Department of General, Visceral and Transplantation Surgery, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Daniel
D
Seehofer
Department of General, Visceral and Transplantation Surgery, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
Georg
G
Damm
D000082:Acetaminophen / Q000008:administration & dosage*; D000328:Adult; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage; D002470:Cell Survival; D016207:Cytokines / Q000378:metabolism; D004008:Diclofenac / Q000008:administration & dosage*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007107:Immune System; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000187:drug effects* / Q000293:injuries* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016207:Cytokines; D017382:Reactive Oxygen Species; D004008:Diclofenac; D000082:Acetaminophen