title: Detection of the mechanism of immunotoxicity of cyclosporine A in murine in vitro and in vivo models.
abstract: Transcriptomics in combination with in vitro cell systems is a powerful approach to unravel modes of action of toxicants. An important question is to which extent the modes of action as revealed by transcriptomics depend on cell type, species and study type (in vitro or in vivo). To acquire more insight into this, we assessed the transcriptomic effects of the immunosuppressive drug cyclosporine A (CsA) upon 6 h of exposure of the mouse cytotoxic T cell line CTLL-2, the thymoma EL-4 and primary splenocytes and compared these to the effects in spleens of mice orally treated with CsA for 7 days. EL-4 and CTLL-2 cells showed the highest similarities in response. CsA affected many genes in primary splenocytes that were not affected in EL-4 or CTLL-2. Pathway analysis demonstrated that CsA upregulated the unfolded protein response, endoplasmic reticulum stress and NRF2 activation in EL-4 cells, CTLL-2 cells and primary mouse splenocytes but not in mouse spleen in vivo. As expected, CsA downregulated cell cycle and immune response in splenocytes in vitro, spleens in vivo as well as CTLL-2 in vitro. Genes up- and downregulated in human Jurkat, HepG2 and renal proximal tubular cells were similarly affected in CTLL-2, EL-4 and primary splenocytes in vitro. In conclusion, of the models tested in this study, the known mechanism of immunotoxicity of CsA is best represented in the mouse cytotoxic T cell line CTLL-2. This is likely due to the fact that this cell line is cultured in the presence of a T cell activation stimulant (IL-2) making it more suitable to detect inhibitory effects on T cell activation.
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, P.O. Box 230, 6700 AE, Wageningen, The Netherlands. peter_oss@hotmail.com.|Department of Toxicogenomics, Maastricht University, P.O. Box 616, 6200 MD, Maastricht, The Netherlands. peter_oss@hotmail.com.
authors.forename: P C J
authors.initials: PC
authors.lastname: Schmeits
authors.affiliation: Centre for Health Protection, National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The Netherlands.|Department of Toxicogenetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: M M
authors.initials: MM
authors.lastname: Schaap
authors.affiliation: Centre for Health Protection, National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The Netherlands.|Department of Toxicogenetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Luijten
authors.affiliation: Department of Toxicogenomics, Maastricht University, P.O. Box 616, 6200 MD, Maastricht, The Netherlands.|Research Group Microbiology and Systems Biology, TNO, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: van Someren
authors.affiliation: Research Group Microbiology and Systems Biology, TNO, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Boorsma
authors.affiliation: Department of Toxicogenomics, Maastricht University, P.O. Box 616, 6200 MD, Maastricht, The Netherlands.|Centre for Health Protection, National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: van Loveren
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, P.O. Box 230, 6700 AE, Wageningen, The Netherlands.|Department of Toxicogenomics, Maastricht University, P.O. Box 616, 6200 MD, Maastricht, The Netherlands.
authors.forename: A A C M
authors.initials: AA
authors.lastname: Peijnenburg
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, P.O. Box 230, 6700 AE, Wageningen, The Netherlands. peter.hendriksen@wur.nl.
authors.forename: P J M
authors.initials: PJ
authors.lastname: Hendriksen
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D016572:Cyclosporine / Q000633:toxicity*; D015536:Down-Regulation / Q000187:drug effects; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D020869:Gene Expression Profiling / Q000379:methods; D056945:Hep G2 Cells; D006801:Humans; D007166:Immunosuppressive Agents / Q000633:toxicity*; D019169:Jurkat Cells; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D058767:Protein Unfolding / Q000187:drug effects; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects* / Q000276:immunology; D013945:Thymoma / Q000276:immunology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D007166:Immunosuppressive Agents; D016572:Cyclosporine
keywords: C57BL/6; CTLL-2; Cyclosporine A; Interspecies comparison; Splenocytes; Transcriptomics