Evaluation of the Multi-ImmunoTox Assay composed of 3 human cytokine reporter cells by examining immunological effects of drugs

•We established a luciferase reporter assay system, the Multi-ImmunoTox Assay (MITA).•MITA can evaluate the effects of chemicals on IL-2, IFN-γ, IL-8, and IL-1β promoters.•MITA correctly reflects the change of mRNA of the mother cells and whole blood cells.•The MITA evaluation of drugs corresponds with their published immunological effects.•MITA can present a novel high-throughput approach to detect chemical immunotoxicity.

We established a luciferase reporter assay system, the Multi-ImmunoTox Assay (MITA), to evaluate the effects on key predictive in vitro components of the human immune system. The system is composed of 3 stable reporter cell lines transfected with 3 luciferase genes, SLG, SLO, and SLR, under the control of 4 cytokine promoters, IL-2, IFN-γ, IL-1β, and IL-8, and the G3PDH promoter. We first compared the effects of dexamethasone, cyclosporine, and tacrolimus on these cell lines stimulated with phorbol 12-myristate 13-acetate and ionomycin, or lipopolysaccharides, with those on mRNA expression by the mother cell lines and human whole blood cells after stimulation. The results demonstrated that MITA correctly reflected the change of mRNA of the mother cell lines and whole blood cells. Next, we evaluated other immunosuppressive drugs, off-label immunosuppressive drugs, and non-immunomodulatory drugs. Although MITA did not detect immunosuppressive effects of either alkylating agents or antimetabolites, it could demonstrate those of the off-label immunosuppressive drugs, sulfasalazine, chloroquine, minocycline, and nicotinamide. Compared with the published immunological effects of the drugs, these data suggest that MITA can present a novel high-throughput approach to detect immunological effects of chemicals other than those that induce immunosuppressive effects through their inhibitory action on cell division.