Vermes I, Haanen C, Richel D J, Schaafsma M R, Kalsbeek-Batenburg E, Reutelingsperger C P
Department of Clinical Chemistry, Medical Spectrum Twente, Enschede, The Netherlands.
Acta Haematol. 1997;98(1):8-13. doi: 10.1159/000203546.
It has been reported that cultured peripheral B lymphocytes of chronic lymphocytic leukemia (B-CLL) patients show a high degree of apoptosis (programmed cell death). Till now, no data exist about the occurrence of in vitro apoptosis of normal B and T cells. We measured the amount of apoptosis and secondary necrosis (type 2 necrosis) in B-CLL lymphocytes and in normal peripheral B and T lymphocytes in culture. Observations were made on B-CLL lymphocytes and on normal B and T cells purified by immunomagnetic cell sorting. Apoptosis and secondary necrosis were measured using a recently described sensitive flow-cytometric assay, probing simultaneously for cell surface exposure of phosphatidylserine with the use of FITC-labeled annexin-V and for cell membrane integrity as demonstrated by the exclusion of propidium iodide. The degree of in vitro apoptosis and secondary necrosis of normal B cells appears to be higher than that of normal T cells, and even higher than that of B-CLL cells. The results indicate that cultured mature circulating normal B lymphocytes exhibit a higher cell death rate than normal T cells and B-CLL lymphocytes.
据报道，慢性淋巴细胞白血病（B-CLL）患者培养的外周B淋巴细胞显示出高度的凋亡（程序性细胞死亡）。到目前为止，尚无关于正常B细胞和T细胞体外凋亡发生情况的数据。我们测量了培养的B-CLL淋巴细胞以及正常外周B细胞和T细胞中的凋亡量和继发性坏死（2型坏死）。对B-CLL淋巴细胞以及通过免疫磁珠细胞分选纯化的正常B细胞和T细胞进行了观察。使用最近描述的一种灵敏的流式细胞术测定法测量凋亡和继发性坏死，该方法同时用异硫氰酸荧光素（FITC）标记的膜联蛋白V探测磷脂酰丝氨酸在细胞表面的暴露情况，并通过碘化丙啶的排斥来证明细胞膜的完整性。正常B细胞的体外凋亡和继发性坏死程度似乎高于正常T细胞，甚至高于B-CLL细胞。结果表明，培养的成熟循环正常B淋巴细胞比正常T细胞和B-CLL淋巴细胞表现出更高的细胞死亡率。
