Wyckoff J H, Shannon B T, Petrini M, Merler E
Immunobiology. 1985 Mar;169(2):175-85. doi: 10.1016/S0171-2985(85)80031-0.
Human peripheral blood monocytes were placed on a discontinuous density gradient of bovine serum albumin and fractionated into five subpopulations. Cells from each subpopulation were assayed for spontaneous cytotoxic activity against K562 tumor cells. Immediately following fractionation, monocytes were not cytotoxic. Following incubation for at least 48 hr, monocytes from two layers of the gradients clearly exhibited greater spontaneous cytotoxic activity than all others. The degree of cytotoxicity expressed by cells of these layers was enhanced by the addition of indomethacin and inhibited by prostaglandin E2 (PGE2). Monocytes acquiring spontaneous cytotoxicity did not secrete measurable levels of PGE2 and had increased levels of purine nucleoside phosphorylase after 72 hs of culture in vitro. Surface markers HNK-1 and Mac-1 normally associated with cytotoxic function, were detected on these cells by indirect immunofluorescence at isolation and after culture. The fraction with the greatest cytotoxic activity showed an increase in the proportion of cells displaying reactivity to HNK-1 after culture compared to initial isolation.
将人外周血单核细胞置于牛血清白蛋白的不连续密度梯度上,分离为五个亚群。检测每个亚群细胞对K562肿瘤细胞的自发细胞毒性活性。分级分离后立即检测,单核细胞无细胞毒性。孵育至少48小时后,梯度中两层的单核细胞明显比其他所有单核细胞表现出更强的自发细胞毒性活性。这些层的细胞所表现出的细胞毒性程度可通过添加吲哚美辛而增强,而被前列腺素E2(PGE2)抑制。获得自发细胞毒性的单核细胞在体外培养72小时后不分泌可测量水平的PGE2,且嘌呤核苷磷酸化酶水平升高。通过间接免疫荧光在分离时和培养后在这些细胞上检测到通常与细胞毒性功能相关的表面标志物HNK-1和Mac-1。与初始分离相比,具有最大细胞毒性活性的部分在培养后显示出对HNK-1有反应的细胞比例增加。