ANALYSIS OF T-CELL SUBSETS IN NORMAL ADULTS - COMPARISON OF WHOLE-BLOOD LYSIS TECHNIQUE TO FICOLL-HYPAQUE SEPARATION BY FLOW-CYTOMETRY
ANALYSIS OF T-CELL SUBSETS IN NORMAL ADULTS - COMPARISON OF WHOLE-BLOOD LYSIS TECHNIQUE TO FICOLL-HYPAQUE SEPARATION BY FLOW-CYTOMETRY
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DOI：
10.1016/0022-1759(87)90434-0
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发表时间：
1987-04-02
影响因子：
2.2
通讯作者：
GINNS, LC
中科院分区：
文献类型：
--
作者：
RENZI, P;GINNS, LC
通过免疫荧光和流式细胞术分析外周血T细胞和T细胞亚群通常用于正常个体和某些疾病状态的研究。从粒细胞和红细胞中分离T细胞的方法各不相同。我们试图确定这些程序是否会改变被分析细胞的组成。在这篇文章中，我们报告说，在正常人中，与全血裂解技术相比，Ficoll-Hypaque淋巴细胞分离选择性地降低了OKT 8+亚群（P < 0.01）。OKT 8+亚群的减少导致OKT 4/OKT 8比值的显著增加（P < 0.01）。这种差异似乎是由于在Ficoll-Hypaque上离心期间OKT 8+细胞的选择性损失。使用Ficoll-Hypaque方法，我们还发现与单克隆抗体反应的淋巴细胞的固定比全血裂解法制备的细胞稳定性差。T细胞亚群的测量结果的解释受到这些细胞分离方法的影响。
Analysis of peripheral blood T cells and T cell subsets by immunofluorescence and flow cytometry is employed commonly in studies of normal individuals and in certain disease states. Methods for separating T cells from granulocytes and erythrocytes vary. We have sought to determine whether or not such procedures alter the composition of cells being analyzed. In this communication, we report that, in normal individuals, Ficoll-Hypaque separation of lymphocytes selectively decreases the OKT8+ subset compared to the whole blood lysis techique (P < 0.01). The decrease in the OKT8+ subset results in a significant increase (P < 0.01) in the OKT4/OKT8 ratio. This disparity appears to be due to a selective loss of OKT8+ cells during centrifugation over Ficoll-Hypaque. With the Ficoll-Hypaque method, we have also found that the fixation of lymphocytes reacted with monoclonal antibodies is less stable than cells prepared by the whole blood lysis method. The interpretation of measurements of T cell subsets is influenced by the method by which these cells are isolated.