Cloning and characterization of guinea pig interleukin-8 receptor
Cloning and characterization of guinea pig interleukin-8 receptor
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DOI：
10.1016/s0006-2952(03)00459-3
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发表时间：
2003-10-01
影响因子：
5.8
通讯作者：
Paquet, JL
中科院分区：
文献类型：
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作者：
Catusse, J;Faye, P;Paquet, JL
cxc趋化因子受体1和2及其配体（CXCL 1、2、3、5、6、7和8）在炎症期间诱导中性粒细胞的选择性募集。这种受体尚未在豚鼠（一种令人感兴趣的动物炎症模型）中被表征。我们报道了鉴定、克隆和鉴定豚鼠CXCL8受体。人CXCL8在豚鼠中性粒细胞体内产生嗜中性粒细胞、趋化性和细胞内钙释放。研究了该受体在嗜中性粒细胞表面的表达。从豚鼠中性粒细胞中克隆了编码功能性CXCL8受体的cDNA并对其进行了测序。采用RT-PCR技术合成该蛋白，引物来自已发表的CXCL8受体的保守区域。该序列为开放阅读框，编码352个氨基酸，在氨基酸水平上与人和兔CXCR2分别有70%和69%的同源性。该受体主要在中性粒细胞中表达，但在肾、肺、脾中也存在，在心脏中也有少量表达。克隆受体转染的细胞显示，该受体对人CXCL8具有高亲和力，略低于与豚鼠中性粒细胞的亲和力。兔和人的CXC趋化因子在转染的细胞中诱导肌醇磷酸积累。受体结合和激活特性以及序列同源性表明，我们鉴定了一种与人类CXCR2受体等效的豚鼠。(C) 2003 Elsevier Inc.版权所有。
CXC-chemokine receptors 1 and 2 and their ligands (CXCL 1, 2, 3, 5, 6, 7, and 8) induce the selective recruitment of neutrophils during inflammation. Such receptors have not been characterized yet in guinea pig, an animal inflammation model of interest. We report the identification, cloning, and characterization of a CXCL8 receptor in guinea pig. Human CXCL8 produced in vivo neutrophilia, chemotaxis and intracellular calcium release of guinea pig neutrophils. The expression of this receptor at their neutrophil surface was investigated. The cDNA encoding a functional CXCL8 receptor was cloned from guinea pig neutrophils and sequenced. It was synthesized using RT-PCR, with oligonucleotide primers derived from well conserved regions of published CXCL8 receptors. This sequence presented an open reading frame coding for 352 amino acids and shares, at the amino acid level, 70 and 69% identity with human and rabbit CXCR2, respectively. The receptor was mainly expressed in neutrophils but it was also present in kidney, lung, spleen and, to a less extent, in heart. Cloned receptor transfected cells showed that this receptor displayed high affinity for human CXCL8, slightly lower than the affinity observed with guinea pig neutrophils. CXC chemokines from both rabbit and human were shown to induce inositol phosphate accumulation in these transfected cells. Receptor binding and activation characteristics together with sequence homology suggested that we identified a guinea pig equivalent of the human CXCR2 receptor. (C) 2003 Elsevier Inc. All rights reserved.