TY - JOUR
T1 - Evaluation of the Multi-ImmunoTox Assay composed of 3 human cytokine reporter cells by examining immunological effects of drugs
AU - Kimura, Yutaka
AU - Fujimura, Chizu
AU - Ito, Yumiko
AU - Takahashi, Toshiya
AU - Aiba, Setsuya
N1 - Funding Information:
This work was supported in part by a Health and Labor Sciences Research Grant in Japan and by the Ministry of Economy, Trade and Industry.
PY - 2014/8
Y1 - 2014/8
N2 - 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.
AB - 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.
KW - Alternatives to animals
KW - Immunotoxicity
KW - Reporter assay
UR - http://www.scopus.com/inward/record.url?scp=84897936561&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84897936561&partnerID=8YFLogxK
U2 - 10.1016/j.tiv.2014.02.013
DO - 10.1016/j.tiv.2014.02.013
M3 - Article
C2 - 24603311
AN - SCOPUS:84897936561
SN - 0887-2333
VL - 28
SP - 759
EP - 768
JO - Toxicology in Vitro
JF - Toxicology in Vitro
IS - 5
ER -