Activation and expression of the nuclear factors of activated T cells, NFATp and NFATc, in human natural killer cells: regulation upon CD16 ligand binding.
Activation and expression of the nuclear factors of activated T cells, NFATp and NFATc, in human natural killer cells: regulation upon CD16 ligand binding.
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在人类天然杀伤细胞中，活化T细胞NFATP和NFATC的核因子的激活和表达：CD16配体结合时调节。
DOI：
10.1084/jem.182.3.801
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发表时间：
1995-09-01
期刊：
影响因子：
--
通讯作者：
Perussia B
中科院分区：
文献类型：
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作者：
Aramburu J;Azzoni L;Rao A;Perussia B
在自然杀伤（NK）细胞中，将来自表面受体的信号转导与细胞因子合成的激活偶联的假定因素尚未阐明。我们在这里报告，活化T细胞的核因子（NFATp），环孢菌素A（CsA）的敏感因子，调节几种细胞因子的转录，介导CD 16诱导的细胞因子基因在人类NK细胞的激活。NK细胞中CD 16（Fc γ RIIIA）诱导的细胞因子mRNA表达通过CsA敏感性和Ca（2+）依赖性机制发生。用CD 16配体刺激NK细胞诱导来自这些细胞的核提取物中的NFAT样DNA结合活性，如在电泳迁移率变动测定中检测到的。这种情况在刺激后以快速动力学发生，通过不需要从头蛋白质合成的CsA敏感性和Ca（2+）依赖性机制进行。NK细胞NFAT存在于未刺激细胞的胞质溶胶中，在刺激时迁移到细胞核，并且可以与AP-1结合。据报道，两种不同的分子NFATp和NFATc介导NFAT活性。使用NFATp-和NFATc-特异性抗体的超移位测定的结果表明，在CD 16配体结合后早期的NK细胞活化主要（如果不是排他的话）涉及NFATp，并且蛋白质印迹分析显示其具有与T淋巴细胞相同的电泳迁移率（约120 kD）。NK细胞不组成性表达NFATc，但在用CD 16配体刺激的2小时内在这些细胞中诱导NFATc mRNA积累。然而，使用可获得的识别T细胞NFATc的mAb的超移位测定显示，在检测的任何时间（长达4 h），在CD 16或佛波酯刺激的细胞的核和细胞质提取物中均未检测到NFATc蛋白。这些结果提供了第一个直接证据，表明CsA敏感性转录因子NFATp和NFATc在人NK细胞中表达，并且它们的活化和/或表达可以在原代细胞中通过单一刺激来调节，在NK细胞中的CD 16的情况下，导致NFATp的早期活化和随后诱导的NFATc mRNA的表达。
The putative factors that couple the signal transduction from surface receptors to the activation of cytokine synthesis in natural killer (NK) cells have not been elucidated. We report here that the nuclear factor of activated T cells (NFATp), a cyclosporin A (CsA)-sensitive factor that regulates the transcription of several cytokines, mediates CD16-induced activation of cytokine genes in human NK cells. CD16 (Fc gamma RIIIA)-induced expression of cytokine mRNA in NK cells occurs via a CsA-sensitive and Ca(2+)-dependent mechanism. Stimulation of NK cells with CD16 ligands induces NFAT-like DNA binding activity in the nuclear extracts from these cells, as detected in electrophoretic mobility shift assays. This occurs with fast kinetics after stimulation, via a CsA-sensitive and Ca(2+)-dependent mechanism that does not require de novo protein synthesis. NK cell NFAT is present in the cytosol of nonstimulated cells, migrates to the nucleus upon stimulation, and can associate with AP-1. Two distinct molecules, NFATp and NFATc, have been reported to mediate NFAT activity. The results of supershift assays using NFATp- and NFATc- specific antibodies indicate that NK cell activation early after CD16 ligand binding involves primarily, if not exclusively, NFATp, and Western blot analysis shows that this has the same electrophoretic mobility (approximately 120 kD) as that of T lymphocytes. NK cells do not express NFATc constitutively, but NFATc mRNA accumulation is induced in these cells within 2 h of stimulation with CD16 ligands. However, supershift assays using the available mAb recognizing the T cell NFATc revealed no detectable NFATc protein in nuclear and cytoplasmic extracts from CD16- or phorbol ester-stimulated cells at any time tested, up to 4 h. These results provide the first direct evidence that both CsA-sensitive transcription factors, NFATp and NFATc, are expressed in human NK cells, and that their activation and/or expression can be regulated in primary cells by a single stimulus, that, in the case of CD16 in NK cells, results in early activation of NFATp and subsequently induced expression of NFATc mRNA.