Yamamura Y, Rodriguez N, Schwartz A, Eylar E, Bagwell B, Yano N
Ponce School of Medicine, AIDS Research Program, Puerto Rico 00732.
Cell Mol Biol (Noisy-le-grand). 1995;41 Suppl 1:S121-32.
A new flow cytometric method was developed to quantitatively assess lymphocyte proliferation simultaneously for different subsets. The cells were stained with a fluorescent dye, PKH-26 and were stimulated with mitogens. The fluorescence intensities (FL2) of proliferating cells were measured by flow cytometry; and each subset was identified by the use of a monoclonal antibody (Mab)-fluorescein-isothiocyanate (FITC) (FL1). FL2 histograms were then analyzed by the cell proliferation model based on the ModFit software (Verity). This new method revealed information which could not be obtained by conventional mitogen assays. For example, the CD4+ and the CD4- T-subsets responded to phytohemagglutinin (PHA) quite differently from each other and it was indicated that activation of one population could significantly alter the response of the other. In addition, even within a subset, all activated cells did not proliferate uniformly. Some cells divided only once while others underwent further cellular division during the same time period. The method is, therefore, invaluable for studying the nature and the extent of interactions between different cellular subsets within a culture.
开发了一种新的流式细胞术方法，用于同时定量评估不同亚群淋巴细胞的增殖情况。细胞用荧光染料PKH-26染色，并用有丝分裂原刺激。通过流式细胞术测量增殖细胞的荧光强度（FL2）；每个亚群通过使用单克隆抗体（Mab）-异硫氰酸荧光素（FITC）（FL1）来鉴定。然后基于ModFit软件（Verity）的细胞增殖模型分析FL2直方图。这种新方法揭示了传统有丝分裂原检测无法获得的信息。例如，CD4 +和CD4 - T亚群对植物血凝素（PHA）的反应彼此非常不同，并且表明一个群体的激活可以显著改变另一个群体的反应。此外，即使在一个亚群内，所有活化细胞也并非均匀增殖。一些细胞仅分裂一次，而其他细胞在同一时间段内进行进一步的细胞分裂。因此，该方法对于研究培养物中不同细胞亚群之间相互作用的性质和程度非常有价值。
