Activation of natural killer cells with interleukin 2 (IL-2) and IL-12 increases perforin binding and subsequent lysis of tumour cells
Activation of natural killer cells with interleukin 2 (IL-2) and IL-12 increases perforin binding and subsequent lysis of tumour cells
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DOI：
10.1046/j.1365-2141.2001.02995.x
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发表时间：
2001-09-01
影响因子：
6.5
通讯作者：
Uharek, L
中科院分区：
文献类型：
--
作者：
Lehmann, C;Zeis, M;Uharek, L
自然杀伤(NK)细胞通过胞吐穿孔素，随后与靶细胞膜结合，形成溶孔，从而对多种不同的肿瘤细胞进行杀伤。然而，一些肿瘤细胞对细胞毒性具有抵抗力。利用NK耐药肿瘤细胞系ML-2、MONOMAC-1、RPMI和L540Cy，我们证明了IL-2和IL-12对NK细胞的激活导致这些肿瘤靶点的显著溶解。为了探讨其机制，我们从未激活和IL-2/IL-12激活的NK细胞中分离出细胞毒颗粒，并比较了它们对K562白血病细胞(对NK细胞介导的杀伤敏感)和ML-2白血病细胞(对NY耐药，细胞介导的溶解)的杀伤作用。与易被NK细胞颗粒杀伤的K562细胞不同，ML-2细胞对来自未激活的NK细胞的颗粒具有抵抗力。然而，IL-2和IL-12激活的墨汁细胞颗粒能够诱导显著的肿瘤细胞溶解。抗穿孔素抗体可完全阻断活化的NIC细胞颗粒对K562和ML-2细胞的杀伤作用，表明穿孔素是NK颗粒诱导细胞死亡的主要原因。比较未激活和IL-2/IL-12激活的NIC细胞的颗粒，ML-2细胞死亡增加是由于穿孔素与靶细胞膜的结合增强所致。然而，功能分析表明，穿孔素结合的差异并不是由于激活的NK细胞增加了穿孔素的产生。我们的结论是，NK细胞的激活导致穿孔素的结合增加，从而导致肿瘤细胞的溶解。
Natural killer (NK) cells can lyse a variety of different tumour cells by exocytosis of perforin, subsequent binding of perforin to the target cell membrane and formation of lytic pores. Some tumour cells, however, are resistant to cellular cytotoxicity. Using the NK-resistant tumour cell lines ML-2, MONOMAC-1, RPMI and L540Cy, we demonstrated that activation of NK cells with interleukin 2 (IL-2) and IL-12 resulted in significant lysis of these tumour targets. To investigate the underlying mechanisms, we isolated the cytotoxic granules from non-activated and IL-2-/IL-12-activated NK cells and compared the killing of K562 leukaemia cells (sensitive to NK cell-mediated lysis) and ML-2 leukaemia cells (resistant to NY, cell-mediated lysis). In contrast to K562 cells, which were easily killed by NK-cell granules, ML-2 cells were resistant to granules from non-activated NK cells. However, granules from INK cells activated with IL-2 and IL-12 were able to induce significant tumour cell lysis. Cell death of both K562 and ML-2 cells by granules from activated NIC cells was completely blocked by anti-perforin antibodies, indicating that perforin mainly accounts for the lysis induced by NK granules. Comparing granules from non-activated and IL-2-/IL-12-activated NIC cells, the increased cell death of ML-2 cells was caused by an improved binding of perforin to the target cell membrane. Functional assays, however, indicated that the differences in perforin binding were not as a result of an augmented production of perforin by activated NK cells. We conclude that activation of NK cells results in an increased binding of perforin and subsequent lysis of tumour cells.