流式细胞仪检测大鼠外周血T细胞亚群(微量全血法)

本实验利用MRC W/OX系列单克隆抗体和流式细胞仪(Epics C型)首次建立了微量全血检测大鼠外周血T细胞亚群的方法,并报告了正常成鼠外周血T细胞亚群分布(n=60):OX-19+细胞占65.45±5.98％(SD),W3/25~+细胞占47.17±6.45％,OX-8~+细胞占24.26±6.18％。W3/25~+亚群和OX-8~+亚群比值为2.12±0.11(SEM)。部分标本流式细胞仪分析结果和荧光显微镜的结果无明显差异(P>0.05)。在抗体染色条件方面,发现当温育温度过高(30℃)时,染色阳性细胞率有明显下降(P<0.001)。实验证实,用微量全血染色不仅简便、可靠、重复性好,且用血量少。同时避免了常规Ficoll分离法时种种弊病。因此用本法可动态观察大鼠外周血T细胞亚群的变化,为进一步研究在药理、病理和免疫学实验中的细胞免疫功能变化提供了方便。

ANALYSES OF RAT PERIPHERAL BLOOD LYMPHOCYTE SUBSETS BY MONOCLONAL ANTIBODIES AND FLOW CYTOMETER(MICRO-VOLUME WHOLE BLOOD METHOD)

In this report,a method by means of McAbs(MRC W/OX)and Flow Cytometer(Epics C)to measure the rnt circulating lymphocyte subsets with microvolume whole blood stain was described.T cell subsets in the peripheral blood of W line adult rats(n=60)thus measured were 65.45±5.98% OX-19+ cells.47.17±6.45% W3/25+ cells and 24.26± 6.18% OX-8+ cells.W3/25+/OX-8+ subsets ratio was 2.12±0.11(SEM).Compared with fluorescein staining method there was no significant difference(P>0.05).In exploring the staining condition,it was found that when incubating temperature augmented above the limited range,for instance,up to 30℃,the percentage of positive staining cells was much lower than that at 4℃ or 10-15℃(P<0.001).It is evident that this simple and rapid method is easy to operate and able to avoid the disadvantage of conventional Ficoll lymphocyte separation method.The method described here makes it possible to observe the functional change of rat pheripheral T cell subsets in the experimention successively.