Comparison of the Pharmacological Efficacies of Immunosuppressive Drugs Evaluated by the ATP Production and Mitochondrial Activity in Human Lymphocytes

Cell Medicine

Hiroshi Satoh¹  Akira Toyama¹  Hiroyasu Sasahara¹  Mahoto Tsukaguchi¹  Kazuya Isogai¹  Kentaro Sugiyama²  Sachiko Tanaka²  Kenji Onda²  Toshihiko Hirano²  Kazuhide Saitoh³  Yuki Nakagawa³  Kota Takahashi³ 

[1] *Division of Pharmacy, Niigata University Medical and Dental Hospital, Niigata, Japan;†Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Japan;‡Division of Urology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan

关键词: CellTiter-Glo assay;    Lymphocyte immunosuppressant sensitivity test (LIST);    3-(4;    5-Dimethylthiazol-2-yl)-2;    5-diphenyltetrazolium bromide (MTT) assay;    Peripheral blood mononuclear cells (PBMCs);   

DOI  :  10.3727/215517913X674207

学科分类：生物科学（综合）

来源: Cognizant Communication Corporation

【 摘 要 】

The lymphocyte immunosuppressant sensitivity test (LIST) using patient peripheral lymphocytes can predict the therapeutic efficacy of immunosuppressive drugs used in renal transplantation. We have evaluated the pharmacological efficacy of drugs by using the LIST with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which measures the cellular mitochondrial activity. The LIST with the MTT assay requires a relatively large amount of blood. As such, we developed a new assay for examining drug sensitivity with a CellTiter-Glo assay, which measures the amount of cellular ATP to help increase the assay's sensitivity and reduce the amount of blood needed. Renal transplant recipients generally receive either cyclosporine or tacrolimus, in addition to mycophenolate mofetil and methylprednisolone, as an immunosuppressive therapy to prevent acute rejection. We evaluated the pharmacological efficacy of these immunosuppressive agents with both the MTT and CellTiter-Glo assays using the peripheral blood mononuclear cells of 21 healthy volunteers. Furthermore, we also examined the relationship between these immunosuppressive agents' pharmacological efficacy and the results of the MTT and CellTiter-Glo assays. The IC50 values for cyclosporine, tacrolimus, mycophenolic acid, and methylprednisolone were significantly correlated between the MTT and CellTiter-Glo assays. The amount of blood cells required for LIST with the CellTiter-Glo assay was able to be reduced to 25% of the amount required for the previously established LIST with the MTT assay procedure. We concluded from these observations that the LIST with the CellTiter-Glo assay should be used instead of the MTT assay for carrying out individualized immunosuppressive therapy in renal transplantation patients.

【 授权许可】

CC BY