In-vivo extravasation induces the expression of interleukin 1 receptor type 1 in human neutrophils
In-vivo extravasation induces the expression of interleukin 1 receptor type 1 in human neutrophils
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DOI：
10.1111/j.1365-2249.2011.04548.x
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发表时间：
2012-04-01
影响因子：
4.6
通讯作者：
Lundahl, J.
中科院分区：
文献类型：
--
作者：
Paulsson, J. M.;Moshfegh, A.;Lundahl, J.
为了解决炎症过程中中性粒细胞的激活问题，我们评估了体内外渗后白细胞介素 1 受体 1 型 (IL-1R1) 的表达。通过皮肤室技术从 11 名健康研究受试者中收集外渗的中性粒细胞，并与外周血中的中性粒细胞进行比较。通过微阵列、定量聚合酶链反应（qPCR）、蛋白质印迹、流式细胞术、酶联免疫吸附测定（ELISA）和免疫电子显微镜（iEM）评估IL-1R1的表达。基因阵列和 qPCR 均证明 IL-1R1 在外渗后被诱导。蛋白质印迹显示外渗白细胞中 IL-1R1 的表达增加。通过流式细胞术和 iEM 在中性粒细胞中进一步证实了这一点，还证明细胞内 IL-1R1 库增加，可通过 N-甲酰基-甲硫氨酸-亮氨酸-苯丙氨酸 (fMLP) 动员。用IL-1刺激外周中性粒细胞导致NFκB和许多下游趋化因子的转录，并且在体内外渗后也诱导了相应的趋化因子。目前的结果表明，IL-1R1 在外渗后被诱导，并存在于中性粒细胞表面以及移动的细胞内库中。此外，中性粒细胞表达功能性 IL-1R1，如 IL-1 刺激后趋化因子的诱导所证明的。结果表明 IL-1 在炎症部位中性粒细胞激活中具有潜在作用。
In order to address neutrophil activation during inflammation we assessed the expression of interleukin 1 receptor type 1 (IL-1R1) following in-vivo extravasation. Extravasated neutrophils were collected from 11 healthy study subjects by a skin chamber technique and compared to neutrophils in peripheral blood. Expression of IL-1R1 was assessed by microarray, quantitative polymerase chain reaction (qPCR), Western blot, flow cytometry, enzyme linked immunosorbent assay (ELISA) and immunoelectron microscopy (iEM). IL-1R1 was induced following extravasation, demonstrated by both gene array and qPCR. Western blot demonstrated an increased expression of IL-1R1 in extravasated leucocytes. This was confirmed further in neutrophils by flow cytometry and iEM that also demonstrated an increased intracellular pool of IL-1R1 that could be mobilized by N-formyl-methionine-leucine-phenylalanine (fMLP). Stimulation of peripheral neutrophils with IL-1 resulted in transcription of NF?B and a number of downstream chemokines and the corresponding chemokines were also induced following in-vivo extravasation. The present results demonstrate that IL-1R1 is induced following extravasation and exists on the neutrophil surface, as well as in a mobile intracellular pool. Furthermore, neutrophils express functional IL-1R1 as demonstrated by the induction of chemokines following IL-1 stimulation. The results indicate a potential role for IL-1 in the activation of neutrophils at inflammatory sites.