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本文探索了应用荧光分光光度计定量测定正常人外周血单个核细胞介素-2受体(IL_2R)的方法及其条件.以PHA与ConA诱导单个核细胞IL_2R表达的最适细胞浓度为3×10~6/ml:PHA和ConA最适浓度分别为100μm/ml和10μg/ml,PHA诱导较ConA为优.PHA诱导细胞IL_2R的表达24h达到高峰,尔后逐渐下降.第一抗体(抗—Tao)和第二抗体(荧光抗体)与单个核细胞孵育时间分别以20min和10min为好.检测同一标本不同复管的细胞(1×10~6)IL_2R结合荧光抗体mol(简称M)为5.22±0.45×10~(10),变异系数(CV)为8.6％,表明该法稳定性较好.测定5例正常人细胞(1×10~6)IL_2R结合荧光抗体M为4.8±0.97×10~(10).CV为20.2％,结果表明该法能较客观检测出不同个体IL_2R表达.
In this paper, we explored the method and the conditions for the quantitative determination of IL-2R in peripheral blood mononuclear cells (PBMCs) using fluorescence spectrophotometer.The optimum cell concentration of IL-2R induced by PHA and ConA was 3 × The optimal concentration of PHA and ConA was 100μm / ml and 10μg / ml, respectively, which was better than that of ConA.PHA induced the expression of IL-2R to peak at 24h, and then decreased gradually.After the first antibody (anti-Tao ) And the second antibody (fluorescent antibody) and mononuclear cell incubation time were 20min and 10min, respectively.It was found that the number of cells (1 × 10 ~ 6) and the fluorescence antibody mol (M) 0.45 × 10 ~ (10), and the coefficient of variation (CV) was 8.6%, indicating that the method has good stability.The determination of IL-1R in 5 normal human cells (1 × 10 ~ 6) was 4.8 ± 0.97 × 10 ~ (10) .CV was 20.2%, the results showed that the method can more objectively detect IL 2R expression in different individuals.