Molecular and biological characterization of the murine leukotriene B4 receptor expressed on eosinophils.
Molecular and biological characterization of the murine leukotriene B4 receptor expressed on eosinophils.
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DOI：
10.1084/jem.188.6.1063
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发表时间：
1998-09-21
期刊：
影响因子：
--
通讯作者：
Luster AD
中科院分区：
文献类型：
--
作者：
Huang WW;Garcia-Zepeda EA;Sauty A;Oettgen HC;Rothenberg ME;Luster AD
白细胞进入组织的运动是由局部产生的化学介质调节的，这些介质统称为化学引诱物。虽然化学引诱物构成了一系列不同的分子，包括蛋白质，肽和脂质，他们似乎都通过一个相关的家庭的七个跨膜G蛋白偶联受体的信号白细胞。嗜酸性粒细胞是一种有效的促炎细胞，在过敏性炎症、寄生虫感染和某些恶性肿瘤期间被吸引到组织中。由于控制嗜酸性粒细胞招募的分子机制还不完全清楚，我们进行了简并聚合酶链反应从小鼠嗜酸性粒细胞分离的cDNA，以确定新的趋化受体。我们报告的分离的cDNA编码的351个氨基酸的糖蛋白，是78%相同的人类基因，已被报道为嘌呤受体（P2Y7）和白三烯B4（LTB4）受体（BLTR）。用该cDNA转染的中国仓鼠卵巢（CHO）细胞特异性结合[3H] LTB 4，解离常数为0.6 ± 0.1 nM。此外，LTB4在转染的CHO细胞中诱导剂量依赖性的细胞内钙流。相反，[35S]dATP不特异性结合这些转染子。该mRNA在暴露于白细胞介素5的嗜酸性粒细胞中以高水平表达，引发腹膜巨噬细胞和中性粒细胞，并且在较小程度上干扰素γ刺激巨噬细胞。在未攻击小鼠的肺、淋巴结和脾中检测到低水平的表达。Western blot分析检测到小鼠嗜酸性粒细胞和肺泡巨噬细胞以及人嗜酸性粒细胞中的mBLTR蛋白。此外，在与嗜酸性粒细胞流入一致的时间过程中，在过敏性肺部炎症的鼠模型中的小鼠肺中发现mBLTR mRNA水平升高。我们的研究结果表明，这种小鼠受体是LTB4受体，在活化的白细胞，包括嗜酸性粒细胞上高度表达，并可能在介导嗜酸性粒细胞募集到炎症灶中发挥重要作用。
The movement of leukocytes into tissues is regulated by the local production of chemical mediators collectively referred to as chemoattractants. Although chemoattractants constitute a diverse array of molecules, including proteins, peptides, and lipids, they all appear to signal leukocytes through a related family of seven transmembrane–spanning G protein–coupled receptors. The eosinophil is a potent proinflammatory cell that is attracted into tissues during allergic inflammation, parasitic infection, and certain malignancies. Since the molecular mechanisms controlling eosinophil recruitment are incompletely understood, we performed a degenerate polymerase chain reaction on cDNA isolated from murine eosinophils to identify novel chemoattractant receptors. We report the isolation of a cDNA that encodes a 351–amino acid glycoprotein that is 78% identical to a human gene that has been reported to be a purinoceptor (P2Y7) and a leukotriene B4 (LTB4) receptor (BLTR). Chinese hamster ovary (CHO) cells transfected with this cDNA specifically bound [3H]LTB4 with a dissociation constant of 0.6 ± 0.1 nM. Furthermore, LTB4 induced a dose-dependent intracellular calcium flux in transfected CHO cells. In contrast, [35S]dATP did not specifically bind to these transfectants. This mRNA was expressed at high levels in interleukin 5–exposed eosinophils, elicited peritoneal macrophages and neutrophils, and to a lesser extent interferon γ stimulated macrophages. Low levels of expression were detected in the lung, lymph node, and spleen of unchallenged mice. Western blot analysis detected the mBLTR protein in murine eosinophils and alveolar macrophages as well as human eosinophils. In addition, elevated levels of mBLTR mRNA were found in the lungs of mice in a murine model of allergic pulmonary inflammation in a time course consistent with the influx of eosinophils. Our findings indicate that this murine receptor is an LTB4 receptor that is highly expressed on activated leukocytes, including eosinophils, and may play an important role in mediating eosinophil recruitment into inflammatory foci.
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DOI：
10.1084/jem.184.4.1483
发表时间：
1996-10-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.8
作者：
Akbar, GKM;Dasari, VR;Kunapuli, SP
通讯作者：
Kunapuli, SP
影响因子：
14.2
作者：
BOCHNER, BS;SCHLEIMER, RP
通讯作者：
SCHLEIMER, RP
影响因子：
2.9
作者：
BOMALASKI, JS;MONG, S
通讯作者：
MONG, S
影响因子：
15.9
作者：
Heath, H;Qin, SX;Mackay, CR
通讯作者：
Mackay, CR