T细胞亚群CD4测定方法的研究

Study on Method of Determining CD4 on T Lymphocyte

为比较外周血T淋巴细胞亚群CD4不同测定方法的差别 ,以流式细胞术为定量手段 ,测定了猴外周血中三种不同方法处理后CD4的表达 .结果表明 :先标后溶法———先用异硫氰荧光素标记的单克隆抗体 (FITC CD4McAb)标记后 ,再加入红细胞溶解液溶掉红细胞的处理方法 ,结果基本等同于传统的淋巴细胞分离法 ,但样本用量仅为传统方法的 1/ 5 ,且操作简单 .激光共焦显微术的形态学研究也证实 :先标后溶法与淋巴细胞分离法相似 ,其细胞膜表面荧光标记清晰 。

To investigate the differentiation of pre preparation method to determine CD4 on T lymphocyte in blood, CD4 expression on T lymphocyte of monkey was determined by the flow cytometrey as a quantitative analysis with three different pre preparation methods. These results showed that (b) method (whole blood was labeled by FITC CD4 McAb. Then red blood cells were dissolved by erythrocytolysis ) was better (a) method (red blood cells were dissolved by erythrocytolysis in whole blood. Then it was labeled by FITC CD4 McAb). CD4 results with (b) method were equal to (c) method (traditional method of lymphocyte separated ). But the volume of sample by (b) method only was 1/5 of (c) method. The pre preparation of (b) method also was sampler than (c) method.Morphologic study by laser scanning confocal microscopy showed (b) method was similar to (c) method. Both of them had a clear fluorescent labeled on the cell membrane but not in the cell.