Nitric oxide inhibits the secretion of T-helper 1- and T-helper 2-associated cytokines in activated human T cells
Nitric oxide inhibits the secretion of T-helper 1- and T-helper 2-associated cytokines in activated human T cells
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DOI：
10.1046/j.1365-2567.1997.00161.x
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发表时间：
1997-02-01
期刊：
影响因子：
6.4
通讯作者：
Neumann, C
中科院分区：
文献类型：
--
作者：
Bauer, H;Jung, T;Neumann, C
调节辅助性T细胞1（Th 1）和辅助性T细胞2（Th 2）免疫应答的平衡的机制是非常感兴趣的，因为它们可以决定过敏性和感染性疾病的结果。最近，在小鼠中，一氧化氮（NO），一个强大的炎症调节剂，已被报道优先下调Th 1介导的免疫反应。在本研究中，我们研究了NO对活化的人T细胞和人T细胞克隆产生Th 1和Th 2相关细胞因子的影响。在NO供体试剂3-morpholinosydnonimine（SIN-1）和S-亚硝基-N-乙酰青霉胺（SNAP）的存在下测量细胞因子分泌。两种NO供体均显著抑制抗CD 3激活的T细胞释放干扰素-γ（IFN-γ）、白细胞介素-2（IL-2）、IL-5、IL-10和IL-4。没有观察到对Th 1相关细胞因子的优先抑制。既没有亚硝酸盐被发现在活化的T细胞的上清液中，也不是特异性mRNA的诱导型和组成型NO合酶检测，表明T细胞本身并没有贡献所观察到的效果的NO供体。抗CD 28单克隆抗体（mAb）共刺激阻止SIN-1/SNAP介导的细胞因子产生的下调只有部分。相反，当T细胞被佛波酯和离子霉素刺激时，它们对SIN-1诱导的细胞因子产生的抑制是难治的。当SIN-1加入后，抗CD 3刺激的发病，抑制作用被认为是不太明显，表明SIN-1可能会干扰早期信号转导事件。加入超氧化物歧化酶（SOD）和过氧化氢酶没有恢复SIN-1的作用，表明细胞因子的抑制是由于NO而不是氧中间体。此外，8-Br-cGMP介导的细胞内cGMP的增加引起与SIN-1和SNAP观察到的相同的细胞因子抑制模式。使用单细胞测定，这些试剂显示出降低产生IFN-γ的T细胞的频率，这表明并非所有T细胞都对SIN-1/SNAP敏感。然而，通过纯化的T细胞亚群（CD 4（+）、CD 8（+）、CD 45 RA（+）和CD 45 RO（+））产生的细胞因子同样受到NO供体的损害。总之，与鼠系统相反，我们的研究结果没有提供证据表明NO在体外优先抑制活化的人T细胞的Th 1-细胞因子分泌。
Mechanisms regulating the balance of T-helper 1 (Th1) and T-helper 2 (Th2) immune responses are of great interest as they may determine the outcome of allergic and infectious diseases. Recently, in mice, nitric oxide (NO), a powerful modulator of inflammation, has been reported to preferentially down-regulate Th1-mediated immune responses. In the present study, we investigated the effect of NO on the production of Th1- and Th2-associated cytokines by activated human T cells and human T-cell clones. Cytokine secretion was measured in the presence of the NO-donating agents 3-morpholinosydnonimine (SIN-1) and S-nitroso-N-acetylpenicillamine (SNAP). Both NO-donors markedly inhibited the release of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-5, IL-10 and IL-4 by anti-CD3 activated T cells. A preferential inhibition of Th1-associated cytokines was not observed. Neither was nitrite found in the supernatants of activated T cells, nor was specific mRNA for inducible and constitutive NO synthase detectable, indicating that T cells themselves did not contribute to the observed effect of the NO donors. Costimulation with anti-CD28 monoclonal antibodies (mAb) prevented SIN-1/SNAP-mediated down-regulation of cytokine production only in part. In contrast, when T cells were stimulated by phorbol-ester and ionomycin, they were refractory to SIN-1-induced inhibition of cytokine production. When SIN-1 was added after the onset of anti-CD3 stimulation, the inhibitory effect was found to be less pronounced, indicating that SIN-1 may interfere with early signal transduction events. The addition of superoxide dismutase (SOD) and catalase did not restore the effects of SIN-1, demonstrating that the inhibition of cytokines was due to NO and not to oxygen intermediates. Furthermore, 8-Br-cGMP-mediated increase of intracellular cGMP caused the same pattern of cytokine inhibition as observed with SIN-1 and SNAP. Using a single cell assay, these agents were shown to reduce the frequency of IFN-gamma-producing T cells, suggesting that not all T cells are susceptible to SIN-1/SNAP. However, cytokine production by purified T-cell subpopulations (CD4(+), CD8(+), CD45RA(+), and CD45RO(+)) was equally impaired by NO donors. In conclusion, in contrast to the murine system, our results do not provide evidence that NO preferentially inhibits Th1-cytokine secretion of activated human T cells in vitro.