人白介素2体外诱生与检测方法的建立  

Establishment of the Method to Induce and Measure Human IL-2 in vitro

作　　者：李桢[1] 张文琳[2] 唐斯[1] 程曦[1] 程良红[1] 张印则[1] 

机构地区：[1]深圳市血液中心,广东深圳518035 [2]唐山钢铁股份有限公司医院职业病科,河北唐山010000

出　　处：《中国实验血液学杂志》2008年第5期1162-1164,共3页Journal of Experimental Hematology

基　　金：深圳市科技计划项目;编号200603201

摘　　要：本研究旨在建立流式磁珠免疫荧光方法,并通过Luminex 100多功能液相分析平台对细胞培养上清液中的IL-2进行定量分析。用密度梯度离心法从ACD抗凝人外周血中分离淋巴细胞,用植物凝集素刺激其分化,在48小时后收集培养上清液,于-20℃冻存待测。用Luminex 100多功能液相分析平台检测IL-2标准品以及样品的相关荧光单位值(RFU),通过标准曲线求出样本中IL-2浓度。结果表明:得出标准品系列的回归方程为Lg(RFU)=1.547+0.867LgC。方差分析显示F=301.7427,p<0.05(ν=6);回归系数t检验显示,t=17.3707(ν=6),p<0.05。结论:建立了人IL-2体外诱生和检测的方法。该方法所得标准曲线有统计学意义。细胞上清液中的IL-2含量(对数)与荧光强度(对数)之间有直线关系。This study was aimed to establish the quantitative analysis of hlL-2 in culture supernatant by multifunctional Luminex 100. The lymphocytes were separated from ACD-anticoagulated peripheral blood by density gradient method. The lymphocytes were stimulated with PHA for 48 hours, and frozen at -20℃. The relative fluorescence units of standard preparations and samples were detected by multifunctional Luminex 100, and the sample concentrations were calculated by standard curve. The results indicated that the regression equation of standard preparation is Lg （RFU） = 1. 547 ＋0. 867 LgC. ANOVA F=301. 7427,p 〈0.05 （v =6）. The analysis of variance showed F = 301. 7427, p 〈 0.05 （ v = 6 ）. The test of regression coefficient showed t = 17. 3707 （ v = 6 ）, p 〈 0.05. It is concluded that method for induction and measurement of human IL-2 in vitro is esteblished. The standard curve established by this way is statistically significant. There is linear relationship between the concentration of hIL-2 and fluorescence intensity.

关 键 词：白细胞介素2 LUMINEX 100 淋巴细胞 植物凝集素 IL-2检测 

分 类 号：Q-331[生物学]