电离辐射与T细胞突变频率的剂量效应关系曲线的建立与验证

目的 建立T细胞受体基因突变频率(T cell receptor mutation frequency,TCR MF)与电离辐射剂量效应关系曲线.方法 招募8名健康成年人,4男4女.采集其外周血,分离淋巴细胞,分装到12孔培养板中,不同剂量(0.00～8.00 Gy)照射后,用植物血凝素蛋白(PHA-P)刺激,加人重组白介素-2(IL-2)培育7d,以流式细胞仪测量TCR MF,拟合剂量-效应曲线.另招募16例不同类型、照射方式及照射剂量的癌症放疗患者,采集外周血,用同样方法分离培养淋巴细胞,测量TCR MF,用拟合的剂量-效应曲线估算肿瘤放疗患者全身等效剂量.采集外周血,观察淋巴细胞染色体双着丝粒和着丝粒环畸变情况,估算受照剂量.结果 TCR MF与电离辐射在0.00～8.00 Gy剂量范围内剂量效应关系良好;大剂量组(2.00 ～8.00 Gy)、小剂量组(0.00～1.00 Gy)及0.00～8.00 Gy剂量组拟合曲线均符合二次多项式模式,拟合方程分别为TCR MF=- 32.8579+ 20.5436D+0.6341D2、TCR MF=1.796+0.017D+ 5.155D2和TCR MF=-0.6229+6.305D+0.6919D2,D为辐射剂量(Gy).用实验建立的曲线估算肿瘤放疗患者的全身受照剂量,与用染色体双着丝粒和着丝粒环畸变率估算的剂量平均相对标准偏差为16.8％.结论 应用TCR基因突变分析技术拟合出的辐射剂量效应曲线,可应用于电离辐射事故受照者近期受照剂量的估算.

Establishment and verification of dose effect relation curves between T cell receptor gene mutation frequency and ionizing radiation dose

Objective To establish the dose-effect curve between TCR MF and ionizing radiation.Methods Peripheral lymphocytes were collected from 8 healthy adults(4 males and 4 females) and cultured in vitro with 12 well culture plates.They were stimulated by PHA-P and IL-2 after exposed to different doses of irradiation (0.00 -8.00 Gy) and cultured for 7 d.The dose-effect curve was established after measuring TCR MF using flow cytometry.Also,using the same method,we separated and cultured the peripheral lymphocytes collected from 16 radiotherapy cancer patients,whose radiation styles and doses were different,and then measured TCR MF to estimate the whole equivalent dose of radiotherapy patients through the dose-effect curve.Peripheral blood was collected and cultured,chromosome aberration (dicentric and ring) was determined under microscope to estimate irradiation dose.Results The relationship of dose-effect between the TCR MF and ionizing radiation (0.00 -8.00 Gy) was well,the curve of large dose group (2.00 -8.00 Gy),low dose group(0.00 - 1.00 Gy) and 0.00 -8.00 Gy dose group were met with the quadratic polynomial model,the equation was TCR MF =-32.8579 + 20.5436D + 0.6341D2,TCR MF =1.796 + 0.017D + 5.155D2 and TCR MF =- 0.6229 + 6.305D + 0.6919D2,respectively.D was the radiation dose ( Gy ).Using the established curve and the chromosome aberration method to estimate the systemic exposure dosage,the average relative deviation was 16.8％.Conclusion The curve established by the TCR gene mutation analysis technology can be applied to exposure dose eatimation of victims in ionization radiation accidents.