PMID- 38045689 OWN - NLM STAT- MEDLINE DCOM- 20231205 LR - 20240325 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 14 DP - 2023 TI - Immuno-inflammatory in vitro hepatotoxicity models to assess side effects of biologicals exemplified by aldesleukin. PG - 1275368 LID - 10.3389/fimmu.2023.1275368 [doi] LID - 1275368 AB - INTRODUCTION: Hepatotoxicity induced by immunotherapeutics is an appearing cause for immune-mediated drug-induced liver injury. Such immuno-toxic mechanisms are difficult to assess using current preclinical models and the incidence is too low to detect in clinical trials. As hepatotoxicity is a frequent reason for post-authorisation drug withdrawal, there is an urgent need for immuno-inflammatory in vitro models to assess the hepatotoxic potential of immuno-modulatory drug candidates. We developed several immuno-inflammatory hepatotoxicity test systems based on recombinant human interleukin-2 (aldesleukin). METHODS: Co-culture models of primary human CD8(+) T cells or NK cells with the hepatocyte cell line HepaRG were established and validated with primary human hepatocytes (PHHs). Subsequently, the HepaRG model was refined by increasing complexity by inclusion of monocyte-derived macrophages (MdMs). The main readouts were cytotoxicity, inflammatory mediator release, surface marker expression and specific hepatocyte functions. RESULTS: We identified CD8(+) T cells as possible mediators of aldesleukin-mediated hepatotoxicity, with MdMs being implicated in increased aldesleukin-induced inflammatory effects. In co-cultures of CD8(+) T cells with MdMs and HepaRG cells, cytotoxicity was induced at intermediate/high aldesleukin concentrations and perforin was upregulated. A pro-inflammatory milieu was created measured by interleukin-6 (IL-6), c-reactive protein (CRP), interferon gamma (IFN-gamma), and monocyte chemoattractant protein-1 (MCP-1) increase. NK cells responded to aldesleukin, however, only minor aldesleukin-induced cytotoxic effects were measured in co-cultures. Results obtained with HepaRG cells and with PHHs were comparable, especially regarding cytotoxicity, but high inter-donor variations limited meaningfulness of the PHH model. DISCUSSION: The in vitro test systems developed contribute to the understanding of potential key mechanisms in aldesleukin-mediated hepatotoxicity. In addition, they may aid assessment of immune-mediated hepatotoxicity during the development of novel immunotherapeutics. CI - Copyright (c) 2023 Roser, Luckhardt, Ziegler, Thomas, Wagner, Damm, Scheffschick, Hewitt, Parnham and Schiffmann. FAU - Roser, Luise A AU - Roser LA AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. FAU - Luckhardt, Sonja AU - Luckhardt S AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. FAU - Ziegler, Nicole AU - Ziegler N AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. FAU - Thomas, Dominique AU - Thomas D AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. AD - pharmazentrum frankfurt/ZAFES, Department of Clinical Pharmacology, Goethe-University Hospital Frankfurt, Frankfurt am Main, Germany. FAU - Wagner, Pia Viktoria AU - Wagner PV AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. FAU - Damm, Georg AU - Damm G AD - Department of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, Leipzig, Germany. FAU - Scheffschick, Andrea AU - Scheffschick A AD - Department of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, Leipzig, Germany. FAU - Hewitt, Philip AU - Hewitt P AD - Chemical and Preclinical Safety, Merck Healthcare KGaA, Darmstadt, Germany. FAU - Parnham, Michael J AU - Parnham MJ AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. FAU - Schiffmann, Susanne AU - Schiffmann S AD - Department of Preclinical Research, Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt am Main, Germany. AD - pharmazentrum frankfurt/ZAFES, Department of Clinical Pharmacology, Goethe-University Hospital Frankfurt, Frankfurt am Main, Germany. AD - Fraunhofer Cluster of Excellence Immune-Mediated Diseases (CIMD), Frankfurt am Main, Germany. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20231117 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - M89N0Q7EQR (aldesleukin) RN - 0 (Interleukin-2) RN - 0 (Biological Products) SB - IM MH - Humans MH - Interleukin-2/pharmacology MH - *Biological Products MH - CD8-Positive T-Lymphocytes MH - *Drug-Related Side Effects and Adverse Reactions MH - *Chemical and Drug Induced Liver Injury/etiology PMC - PMC10693457 OTO - NOTNLM OT - immune cells OT - immune-related adverse events OT - immunotherapy OT - in vitro prognostics OT - triple-culture model COIS- MP is an employee of EpiEndo Pharmaceuticals ehf. PH is an employee of Merck KGaA. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2023/12/04 06:42 MHDA- 2023/12/05 17:44 PMCR- 2023/01/01 CRDT- 2023/12/04 04:42 PHST- 2023/08/09 00:00 [received] PHST- 2023/10/27 00:00 [accepted] PHST- 2023/12/05 17:44 [medline] PHST- 2023/12/04 06:42 [pubmed] PHST- 2023/12/04 04:42 [entrez] PHST- 2023/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2023.1275368 [doi] PST - epublish SO - Front Immunol. 2023 Nov 17;14:1275368. doi: 10.3389/fimmu.2023.1275368. eCollection 2023.