Kuzuya Takafumi, Kobayashi Takaaki, Katayama Akio, Nagasaka Takaharu, Miwa Yuko, Uchida Kazuharu, Nakao Akimasa, Yamada Kiyofumi
Department of Hospital Pharmacy, Nagoya University School of Medicine, Japan.
Biol Pharm Bull. 2009 Apr;32(4):604-8. doi: 10.1248/bpb.32.604.
Inhibition of cytokine production is the main immunosuppressive effect of cyclosporine (CsA), which is widely used in organ transplantation. Pharmacodynamic (PD) assay for evaluating the inhibition of interleukin-2 (IL-2) production for each patient could provide a more appropriate dosing regimen. We measured the suppression of IL-2 mRNA expression in whole blood following the addition of a range of CsA concentrations by a real-time reverse transcription-polymerase chain reaction (RT-PCR) method. Individual CsA sensitivity on the IL-2 mRNA expression was assessed with healthy subjects both in vitro and ex vivo. We also evaluated it in pre-transplant patients before taking immunosuppressive drugs. Sigmoid E(max) model was used to analyze the relationship between CsA concentration and IL-2 mRNA expression. The assay was completed within 8 h. The concentration that resulted in IC(50) showed high reproducibility and specificity among the healthy subjects (p<0.005, n=5). Ex vivo study indicated similar inhibition profiles to those of in vitro studies (n=3). The values of IC(50) obtained from patients (n=22) also showed large variations and were significantly lower than those from healthy subjects (p<0.05). Semi-quantitative RT-PCR was considered to be a rapid and reliable assay. Our data imply that measurement of IL-2 mRNA levels in whole blood could be valuable in monitoring CsA PD in transplant patients.
抑制细胞因子产生是环孢素(CsA)的主要免疫抑制作用,CsA在器官移植中广泛应用。评估每位患者白细胞介素-2(IL-2)产生抑制情况的药效学(PD)检测可为制定更合适的给药方案提供依据。我们通过实时逆转录聚合酶链反应(RT-PCR)方法,测定了添加一系列CsA浓度后全血中IL-2 mRNA表达的抑制情况。在体外和体内对健康受试者评估了个体对CsA对IL-2 mRNA表达的敏感性。我们还在移植前未服用免疫抑制药物的患者中进行了评估。采用Sigmoid E(max)模型分析CsA浓度与IL-2 mRNA表达之间的关系。该检测在8小时内完成。导致IC(50)的浓度在健康受试者中显示出高重现性和特异性(p<0.005,n = 5)。体内研究表明与体外研究具有相似的抑制特征(n = 3)。从患者(n = 22)获得的IC(50)值也显示出较大差异,且显著低于健康受试者(p<0.05)。半定量RT-PCR被认为是一种快速且可靠的检测方法。我们的数据表明,检测全血中IL-2 mRNA水平在监测移植患者的CsA药效学方面可能具有重要价值。