Schlesinger M, Lew F, Bekesi J G
J Clin Lab Immunol. 1984 Apr;13(4):195-202.
Subpopulations of NK-cells recovered in various Percoll fractions had similar Leu 7 (HNK-1) antigenicity. Leu 11 paralleled Leu 7 in its expression in the high density Percoll fractions, while cells in the lowest density showed a somewhat higher reactivity with Leu 11 than with Leu 7 antibody. A similar small proportion (20-30%) of NK-cells recovered in all Percoll fractions was susceptible to lysis by treatment with T8 and complement. Exposure to T11 and complement, which eliminate approximately 50% of the NK activity in unfractionated PBL, reduced the cytotoxicity of NK-cells recovered in the high density fraction to a significantly greater extent (60%) than that of those recovered in the low density (30%). Flow microfluorometry analysis using monoclonal anti-T reagents showed that while Leu 4 (pan T-marker) was expressed on all high density T-cells, a subpopulation of low density T-cells did not express this antigen. The proportion of Leu 3a positive (helper T) cells was found to be significantly higher among the high density T-cells than among the low density T-cells. There was a relative enrichment of Leu 2a positive (cytotoxic/suppressor T) cells recovered in the lower density Percoll fractions. Subsets of NK- and T-cells, differing in their density, were thus shown to also differ in their cell surface antigenic markers. Percoll density fractionation may prove to be a valuable tool for separation and analysis of NK- and T-cell subsets.
在不同Percoll组分中回收的NK细胞亚群具有相似的Leu 7(HNK-1)抗原性。Leu 11在高密度Percoll组分中的表达与Leu 7平行,而在最低密度的细胞中,与Leu 11抗体的反应性略高于与Leu 7抗体的反应性。在所有Percoll组分中回收的NK细胞中,有相似的一小部分(20%-30%)对用T8和补体处理敏感。用T11和补体处理可消除未分级外周血淋巴细胞中约50%的NK活性,与低密度组分中回收的NK细胞相比,高密度组分中回收的NK细胞的细胞毒性降低的程度更大(60%),而低密度组分中回收的NK细胞的细胞毒性降低30%。使用单克隆抗T试剂的流式微荧光分析表明,虽然Leu 4(泛T标记)在所有高密度T细胞上表达,但低密度T细胞亚群不表达该抗原。发现Leu 3a阳性(辅助性T)细胞在高密度T细胞中的比例显著高于低密度T细胞。在较低密度的Percoll组分中回收的Leu 2a阳性(细胞毒性/抑制性T)细胞相对富集。因此,密度不同的NK细胞和T细胞亚群在细胞表面抗原标志物方面也存在差异。Percoll密度分级法可能被证明是分离和分析NK细胞和T细胞亚群的一种有价值的工具。