离体诱导淋巴细胞TCR基因突变检测实验条件的探讨  

Study on the experimental conditions of mutation induction of lymphocyte TCR gene in vitro

作　　者：侯殿俊[1] 马娅[1] 刘伟[1] 乔建维[1] 卢峰[1] 商希梅[1] 李洁清[1] 封丽[1] 

机构地区：[1]山东省医学科学院放射医学研究所,山东济南250062

出　　处：《中国职业医学》2008年第6期475-476,479,共3页China Occupational Medicine

基　　金：山东省自然基金项目资助(Y2007C151)

摘　　要：目的完善外周血淋巴细胞TCR基因突变的离体培养方法,为其作为生物剂量计提供试验模型。方法以不同剂量(0～8 Gy)的X射线照射新鲜分离的健康成人周围血淋巴细胞,用不同浓度的植物血凝素(PHA-P)、重组人白细胞介素2(rhIL2)协同刺激,37℃、5%CO2、饱和湿度条件下培养7 d,流式细胞术检测。结果PHA-P及rhIL2协同刺激,细胞的活化增殖要优于PHA-P单一刺激后再加入rhIL2。TCR基因位点突变频率(TCRMF)在0～8 Gy剂量范围内,剂量效应拟和曲线以二次多项式模式相关性最好。结论采用本实验改进的方法,可缩短TCR基因突变表型表达的时间,可为尽快估算受照人员的辐射生物剂量提供理论依据。Objective To consummate the culture method of TCR gene mutation of peripheral blood lymphocytes to provide a test model as biological dosimeter. Methods Peripheral lymphocytes freshly isolated from healthy adult donors were irradiated with X-ray at doses of ranging from 0 to 8 Gy, then cultured with interleukin-2（rhIL2） and phytohemagglutinin（PHA-P） at the conditions of 37℃, 5% CO2 and saturation humidity for7 days. The mutant frequencies of TCR gene （TCRMF） were detected by flow cytometry. Results The activation of cell proliferation by PHA-P and rhIL2 coordinated stimulation was better than that by stimulation of PHA-P followed by rhIL2. TCRMF were found to be optimal fitting the quadratic polynomial dose-response model within the doses of 0-8 Gy. Conclusion This improved method can reduce the time of expression of the TCR gene mutation phenotype. At the same time it can provide a theoretical basis for estimating the biological radiation dose as quickly as possible.

关 键 词：辐射照射 基因突变 TCR基因 生物剂量计 

分 类 号：X591[环境科学与工程—环境工程]