20.500.12556/RUL-182837 Drug repurposing for cytokine storm baricitinib, ciclesonide, and acalabrutinib modulate cytokine release in a translational in vitro lymphoblastoid cell line model Cytokine storm, a life-threatening hyperinflammatory response seen in severe infections like COVID-19, autoimmune diseases, and certain therapies, remains difficult to treat due to limited approved therapeutics. To support preclinical drug development and repurposing in line with 3R principles, we present a novel in vitro model for screening of cytokine-modulating compounds. Herein, we demonstrate that lymphoblastoid cell lines (LCL cells), derived from healthy donors, provide a translationally relevant and phenotypically diverse platform for cytokine profiling and drug screening. Upon stimulation with PMA/ionomycin, LCL cells exhibited a robust secretion of pro-inflammatory (IL-6, IL-8, TNF-α) and anti-inflammatory (IL-10) cytokines, closely mirroring the cytokine profile of activated whole blood. Compared to established immune cell lines THP-1 and Jurkat, LCL cells showed broader cytokine responses and retained interindividual variability. We validated the model for cytokine-modulating drug screening using two well-established immunosuppressants, dexamethasone and cyclosporin A. Dexamethasone reduced IL-6 by 21 %, IL-8 by 71 %, IL-12 by 68 %, TNF-α by 83 %, and increased IL-10 by 51 %. Cyclosporin A lowered IL-6 by 34 %, IL-8 by 69 %, IL-10 by 48 %, and TNF-α by 41 %. Subsequently, we tested a panel of clinically investigated COVID-19 therapies, selected based on a comprehensive literature review. Ciclesonide and acalabrutinib showed strong suppression of all measured cytokines by more than 60 %, while baricitinib, which has become a standard of care for severe COVID-19, reduced IL-6 by 70 %, IL-12 by 68 %, and TNF-α by 47 %, but had minimal effect on IL-8. Notably, LCL cells retained interindividual variability, as demonstrated by diverse cytokine as well as divergent NF-κB nuclear translocation responses, supporting the model's capacity to reveal real-world patient heterogeneity. Collectively, our findings establish LCL cells as a reproducible, immunologically responsive, and patient-representative platform for cytokine storm research and high-content drug testing, providing both mechanistic insight and translational relevance. cytokine storm lymphoblastoid cell lines LCL cells immunomodulatory drug screening drug repurposing interindividual variability citokinska nevihta limfoblastoidne celične linije LCL celice imunomodulatorji presejalni testi za zdravila prenamenitev zdravil medindividualna variabilnost klinična biokemija cepiva proti covidu-19 zdravila true false true Angleški jezik Slovenski jezik Članek v reviji 2026-05-25 13:33:30 2026-05-25 13:33:40 2026-05-26 04:35:53 0000-00-00 00:00:00 2026 0 0 12 str. art. 157083 Vol. 197 Jan. 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616-097:615.2 1096-0023 10.1016/j.cyto.2025.157083 260350979 1-s2.0-S1043466625002303-main.pdf 1-s2.0-S1043466625002303-main.pdf 1 FE91E3216480F44F33566466E7AE6951 cf2ef1ec8dbf05a4bd10dd5f7180136720d1f2f6ac0d0738b9ad4714d6be13f5 fb39ae7a-582d-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234419 https://www.sciencedirect.com/science/article/pii/S1043466625002303 0 3e703c7c-582d-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234417 Fakulteta za farmacijo 0 0 0