Wang Junyu, Fan Hua
Department of Emergency Medicine, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.
Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China. Email:
Zhonghua Yi Xue Za Zhi. 2014 Dec 30;94(48):3809-12.
To explore the role of P70S6 kinase phosphorylation as a downstream of mammalian target of rapamycin (mTOR) pathway in CD3 positive cells of liver transplant patients.
A total of 84 liver transplant recipients were recruited from our hospital and divided into 3 treatment groups of sirolimus (n = 26), tacrolimus (n = 35) and cyclosporine (n = 23). The P70S6 kinase phosphorylation of CD3 positive cell of patients and healthy control (HC) were analyzed by phospho-flow cytometry. A correlation analysis between P70S6 kinase phosphorylation and sirolimus trough level was performed. Intra-individual variability and inter-individual variability of P70S6 kinase phosphorylation were measured.
The P70S6 kinase phosphorylation in HC showed a low degree of intra-individual variability (3.5% to 5.6%) while the inter-individual variability between different healthy volunteers was higher (18.9% to 22.5%). The P70S6 kinase phosphorylation in patients treated with sirolimus (28.9 ± 10.5) was significantly lower than in HC (57.2 ± 8.4, P < 0.001), tacrolimus (42.5 ± 14.1, P < 0.001) or cyclosporine treated ones (51.4 ± 10.9, P < 0.001). The P70S6 kinase phosphorylation in HC (57.2 ± 8.4) was significantly higher than in tacrolimus (42.5 ± 14.1, P < 0.01) or cyclosporine treated patients (51.4 ± 10.9, P < 0.05). No correlation existed between P70S6 kinase phosphorylation and trough level of sirolimus (r = -0.18, P = 0.39).
Phospho-flow cytometry assay has been established for determining the degree of mTOR inhibition by assessing P70S6 kinase phosphorylation. Quantification of P70S6 kinase phosphorylation may play an adjunct role in pharmacodynamically guiding an individualized mTOR inhibitor based immunosuppression.
探讨雷帕霉素哺乳动物靶点（mTOR）通路下游的P70S6激酶磷酸化在肝移植患者CD3阳性细胞中的作用。
从我院招募84例肝移植受者，分为西罗莫司治疗组（n = 26）、他克莫司治疗组（n = 35）和环孢素治疗组（n = 23）。采用磷酸化流式细胞术分析患者及健康对照（HC）CD3阳性细胞的P70S6激酶磷酸化情况。对P70S6激酶磷酸化与西罗莫司谷浓度进行相关性分析。测定P70S6激酶磷酸化的个体内变异性和个体间变异性。
HC中P70S6激酶磷酸化的个体内变异性较低（3.5%至5.6%），而不同健康志愿者之间的个体间变异性较高（18.9%至22.5%）。西罗莫司治疗患者的P70S6激酶磷酸化水平（28.9±10.5）显著低于HC（57.2±8.4，P<0.001）、他克莫司治疗患者（42.5±14.1，P<0.001）或环孢素治疗患者（51.4±10.9，P<0.001）。HC的P70S6激酶磷酸化水平（57.2±8.4）显著高于他克莫司治疗患者（42.5±14.1，P<0.01）或环孢素治疗患者（51.4±10.9，P<0.05）。P70S6激酶磷酸化与西罗莫司谷浓度之间无相关性（r = -0.18，P = 0.39）。
已建立磷酸化流式细胞术检测方法，通过评估P70S6激酶磷酸化来确定mTOR的抑制程度。P70S6激酶磷酸化的定量分析可能在药效学指导基于mTOR抑制剂的个体化免疫抑制中发挥辅助作用。
