Kutvirt S G, Lewis S L, Simon T L
Department of Pathology, University of New Mexico School of Medicine, Biomedical Research Facility, Albuquerque 87131.
Br J Biomed Sci. 1993 Dec;50(4):321-8.
Flow cytometry techniques for immunophenotyping have revolutionised the diagnosis and monitoring of paediatric immunological disorders. Although recent studies in adult subjects discourage the use of density gradients for cell preparation prior to phenotyping, these procedures continue to be used. The purpose of this study was to determine the effect of density gradient separation on lymphocyte phenotypes from neonates, infants, and adults as compared to whole blood determinations. Subset distributions were different with the two procedures. In all three groups, CD19+ (B cell) and CD8+ (suppressor/cytotoxic T cell) percentages were significantly lower and CD3-CD56+ (NK cell) percentages were significantly higher in the density gradient separated cells. The loss of CD8+ cells in density gradient separation was shown to be a selective event. The CD8+CD11b- (cytotoxic T) subset percentages were lower in the density gradient separated cells, while the percentages of CD8+CD11b+ (suppressor T) cells were not affected by separation procedure. Because of the selective loss of lymphocytes on density gradients, the use of a whole blood technique for immunophenotyping in paediatric subjects is recommended.
用于免疫表型分析的流式细胞术彻底改变了儿童免疫性疾病的诊断和监测。尽管最近针对成年受试者的研究不提倡在表型分析前使用密度梯度进行细胞制备，但这些方法仍在使用。本研究的目的是确定与全血检测相比，密度梯度分离对新生儿、婴儿和成人淋巴细胞表型的影响。两种方法的亚群分布不同。在所有三组中，密度梯度分离的细胞中CD19 +（B细胞）和CD8 +（抑制/细胞毒性T细胞）百分比显著降低，而CD3 - CD56 +（NK细胞）百分比显著升高。密度梯度分离中CD8 +细胞的损失被证明是一个选择性事件。密度梯度分离的细胞中CD8 + CD11b -（细胞毒性T）亚群百分比更低，而CD8 + CD11b +（抑制性T）细胞百分比不受分离程序影响。由于密度梯度上淋巴细胞的选择性损失，建议在儿科受试者中使用全血技术进行免疫表型分析。
