Class IA phosphatidylinositide 3-kinases, rather than p110γ, regulate formyl-methionyl-leucyl-phenylalanine-stimulated chemotaxis and superoxide production in differentiated neutrophil-like PLB-985 cells
Class IA phosphatidylinositide 3-kinases, rather than p110γ, regulate formyl-methionyl-leucyl-phenylalanine-stimulated chemotaxis and superoxide production in differentiated neutrophil-like PLB-985 cells
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DOI：
10.4049/jimmunol.176.12.7621
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发表时间：
2006-06-15
影响因子：
4.4
通讯作者：
Naccache, Paul H.
中科院分区：
文献类型：
--
作者：
Boulven, Isaline;Levasseur, Sylvain;Naccache, Paul H.
I类PI 3 K通过形成磷脂酰肌醇-3，4，5-三磷酸盐（PI（3，4，5）P-3）被认为是中性粒细胞对趋化因子反应的基本要素。此外，最近开发的PI 3 K缺陷型小鼠和亚型特异性抑制剂能够检查不同PI 3 K亚型在中性粒细胞活化中的贡献。然而，这些不同研究的结果是相互矛盾的，不同PI 3 K亚型的确切作用尚未明确，特别是在人类细胞中。在本研究中，我们使用了不同的方法来评估不同的PI 3 K亚型的作用，在响应趋化剂fMLP。我们抑制PI 3 K活性的瞬时表达后，核转染的显性阴性突变体的p85 α或p110 γ在人骨髓细胞系PLB-985，它可以被诱导表达嗜中性粒细胞样表型。用这种方法获得的数据表明，由fMLP触发的PI（3，4，5）P-3的产生是双相的，在短时间内观察到完全取决于p110 γ活性的产生峰值，以及由I-A类PI 3 K介导的延迟相。我们还提供了PI 3 K依赖性功能反应（即，超氧化物产生和趋化性）主要涉及PI 3 K I-A，并暗示PI（3，4，5）P-3产生的延迟相，而p110 γ和PI（3，4，5）P-3的早期峰在这些反应的起始或控制中不起主要作用。
Class I PI3Ks, through the formation of phosphatidylinositol-3,4,5-trisphosphate (PI(3,4,5)P-3) are thought of as essential elements of the neutrophil response to chemotactic factors. Moreover, the recent development of PI3K-deficient mice and isoform-specific inhibitors enabled examinations of the contribution of the distinct PI3K isoforms in neutrophil activation. However, the results of these various studies are conflicting, and the exact role of the different PI3K isoforms is not yet clearly established, particularly in human cells. In the present study, we used a different approach to assess the role of the distinct PI3K isoforms in response to the chemotactic agent fMLP. We inhibited PI3K activities by the transient expression following nucleofection of dominant negative mutants of either p85 alpha or p110 gamma in the human myeloid cell line PLB-985, which can be induced to express a neutrophil-like phenotype. The data obtained with this approach showed that the production of PI(3,4,5)P-3 triggered by fMLP is biphasic, with a peak of production observed in a short time period that entirely depends on p110 gamma activity, and a delayed phase that is mediated by class I-A PI3K. We also provide evidence that the PI3K-dependent functional responses (i.e., superoxide production and chemotaxis) induced by the chemotactic factor mainly involve PI3K I-A and, by implication, the delayed phase of PI(3,4,5)P-3 production, whereas p110 gamma and the early peak of PI(3,4,5)P-3 do not play major roles in the initiation or the control of these responses.