title: Successful validation of genomic biomarkers for human immunotoxicity in Jurkat T cells in vitro.
abstract: Previously, we identified 25 classifier genes that were able to assess immunotoxicity using human Jurkat T cells. The present study aimed to validate these classifiers. For that purpose, Jurkat cells were exposed for 6 h to subcytotoxic doses of nine immunotoxicants, five non-immunotoxicants and four compounds for which human immunotoxicity has not yet been fully established. RNA was isolated and subjected to Fluidigm quantitative real time (qRT)-PCR analysis. The sensitivity, specificity and accuracy of the screening assay as based on the nine immunotoxicants and five non-immunotoxicants used in this study were 100%, 80% and 93%, respectively, which is better than the performance in our previous study. Only one compound was classified as false positive (benzo-e-pyrene). Of the four potential (non-)immunotoxicants, chlorantraniliprole and Hidrasec were classified immunotoxic and Sunset yellow and imidacloprid as non-immunotoxic. ToxPi analysis of the PCR data provided insight in the molecular pathways that were affected by the compounds. The immunotoxicants 2,3-dichloro-propanol and cypermethrin, although structurally different, affected protein metabolism and cholesterol biosynthesis and transport. In addition, four compounds, i.e. chlorpyrifos, aldicarb, benzo-e-pyrene and anti-CD3, affected genes in cholesterol metabolism and transport, protein metabolism and transcription regulation. qRT-PCR on eight additional genes coding for similar processes as defined in ToxPi analyzes, supported these results. In conclusion, the 25 immunotoxic classifiers performed very well in a screening with new non-immunotoxic and immunotoxic compounds. Therefore, the Jurkat screening assay has great promise to be applied within a tiered approach for animal free testing of human immunotoxicity.
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.|Department of Toxicogenomics, Maastricht University, the Netherlands.
authors.forename: Peter C J
authors.initials: PC
authors.lastname: Schmeits
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.|Department of Toxicogenomics, Maastricht University, the Netherlands.
authors.forename: Jia
authors.initials: J
authors.lastname: Shao
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
authors.forename: Danique A
authors.initials: DA
authors.lastname: van der Krieken
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
authors.forename: Oscar L
authors.initials: OL
authors.lastname: Volger
authors.affiliation: Department of Toxicogenomics, Maastricht University, the Netherlands.|Centre for Health Protection, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
authors.forename: Henk
authors.initials: H
authors.lastname: van Loveren
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
authors.forename: Ad A C M
authors.initials: AA
authors.lastname: Peijnenburg
authors.affiliation: RIKILT Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
authors.forename: Peter J M
authors.initials: PJ
authors.lastname: Hendriksen
mesh_terms: D000448:Aldicarb / Q000494:pharmacology / Q000633:toxicity; D001391:Azo Compounds / Q000494:pharmacology / Q000633:toxicity; D001580:Benzopyrenes / Q000494:pharmacology / Q000633:toxicity; D054316:Biomarkers, Pharmacological; D002728:Chlorohydrins / Q000494:pharmacology / Q000633:toxicity; D004390:Chlorpyrifos / Q000494:pharmacology / Q000633:toxicity; D005819:Genetic Markers / Q000187:drug effects*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology / Q000633:toxicity; D000922:Immunotoxins / Q000494:pharmacology*; D066298:In Vitro Techniques; D019169:Jurkat Cells / Q000187:drug effects*; D000073943:Neonicotinoids; D009574:Nitro Compounds / Q000494:pharmacology / Q000633:toxicity; D011722:Pyrethrins / Q000494:pharmacology / Q000633:toxicity; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018675:Toxicity Tests; D062367:ortho-Aminobenzoates / Q000494:pharmacology / Q000633:toxicity
chemical_list: D001391:Azo Compounds; D001580:Benzopyrenes; D054316:Biomarkers, Pharmacological; D002728:Chlorohydrins; D005819:Genetic Markers; D007093:Imidazoles; D000922:Immunotoxins; D000073943:Neonicotinoids; D009574:Nitro Compounds; D011722:Pyrethrins; D062367:ortho-Aminobenzoates; C017160:cypermethrin; C082359:imidacloprid; C517733:chlorantranilipole; C026487:benzo(e)pyrene; C076159:2,3-dichloro-1-propanol; D000448:Aldicarb; C005842:6-hydroxy-5-((p- sulfophenyl)azo)-2-naphthalenesulfonic acid disodium salt; D004390:Chlorpyrifos
keywords: Fluidigm; Jurkat; ToxPi; biomarker; classifier; genomic; high throughput; immunotoxicity; prediction; qRT-PCR