Drug Metabolism and Pharmacokinetics Research Laboratories, Research, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, Japan.
Drug Metab Pharmacokinet. 2020 Apr;35(2):207-213. doi: 10.1016/j.dmpk.2019.11.003. Epub 2019 Dec 21.
As the application of flow cytometry to a quantitative pharmacokinetic study with adoptive T cell therapy is new, we aimed to investigate the quantitativity of flow cytometry-based analysis for the pharmacokinetic assessment of circulating human T cells in a preclinical study. We evaluated the selectivity, linearity, accuracy, precision, and sensitivity of flow cytometry-based analysis for human CD8 T cells in immunodeficient mouse blood. The CD3/8/45-positive cell population was successfully distinguished from the negative population. Linear regression analysis for the calibration curve showed good linearity and recovery was approximately 100%. Acceptable inter- and intra-day precision and accuracy were observed and the lower limit of quantification (30 cells/50 μL) was validated with acceptable precision and accuracy. Blood concentrations of human CD8 T cells in immunodeficient mice were then evaluated after administration using this method and the time-concentration profile of human T cells in mice was successfully assessed. The present study is the first to clarify the quantitativity of flow cytometry-based analysis for circulating human T cells in animals. The concept of the present study would be applicable to quantitative pharmacokinetics/efficacy or safety analysis of adoptive T cell therapy.
由于流式细胞术在过继性 T 细胞治疗的定量药代动力学研究中的应用尚属新领域，我们旨在通过临床前研究来探讨流式细胞术分析用于循环人 T 细胞药代动力学评估的定量能力。我们评估了流式细胞术分析在免疫缺陷小鼠血液中循环人 CD8 T 细胞的选择性、线性、准确性、精密度和灵敏度。成功地区分了 CD3/8/45 阳性细胞群和阴性细胞群。校准曲线的线性回归分析显示出良好的线性，回收率接近 100%。观察到可接受的日间和日内精密度和准确性，验证了定量下限（30 个细胞/50 μL）具有可接受的精密度和准确性。然后使用该方法评估了免疫缺陷小鼠中给予人 CD8 T 细胞后的血液浓度，并成功评估了小鼠中人 T 细胞的时间浓度曲线。本研究首次阐明了流式细胞术分析用于动物循环人 T 细胞的定量能力。本研究的概念适用于过继性 T 细胞治疗的定量药代动力学/疗效或安全性分析。
