Lipoxin A(4) receptor activation is distinct from that of the formyl peptide receptor in myeloid cells: Inhibition of CD11/18 expression by lipoxin A(4)-lipoxin A(4) receptor interaction
Lipoxin A(4) receptor activation is distinct from that of the formyl peptide receptor in myeloid cells: Inhibition of CD11/18 expression by lipoxin A(4)-lipoxin A(4) receptor interaction
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DOI：
10.1021/bi00051a016
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发表时间：
1995-12-26
期刊：
影响因子：
2.9
通讯作者：
Serhan, CN
中科院分区：
文献类型：
--
作者：
Fiore, S;Serhan, CN
脂蛋白A（4）（LXA（4））与人类嗜中性粒细胞和分化的HL-60细胞中的高亲和力受体相互作用。最近，我们表征了一个髓样衍生的cDNA，该cDNA编码LXA（4）高亲和力受体（LXA（4）R）[Fiore，S.，Maddox，J.F.，Perez，Perez，H.D。和Serhan，C.N。 （1994）J。Exp。医学180，253-260]早先表示为相关的N-甲梅基肽受体（RFP）。为了检查该受体的选择性，我们测试了其偏爱H-3-LXA（4）与H-3-N-甲基甲基甲基甲基苯基苯胺（H-3-FMLP）的偏爱。当受体转染的中国仓鼠卵巢细胞暴露于H-3-LXA（4）或H-3-FMLP时，LXA（4）的受体亲和力超过了1000倍FMLP（6.1 nm vs 5 MU M ）。分化后，HL-60细胞获得高亲和力结合位点，并对LXA（4）和FMLP响应。使用RFP探针对分化的HL-60细胞进行了北印迹分析显示，在2.1 kb时表现出一个特征带。暴露于RFP反义寡核苷酸的分化HL-60细胞选择性丢失的H-3-LXA（4）结合以及LXA（4）刺激的脂质重塑，与LXA（4）r的mRNA损失相似。相反，仍然观察到FMLP受体，H-3-FMLP特异性结合和FMLP诱导的磷脂酶D活性的特异性mRNA。用在LXA中针对肽升高的抗血清治疗人类嗜中性粒细胞（4）R第三个细胞外结构域也导致选择性废除H-3-LXA（4）与多形核白细胞（PMN）的特异性结合，而无需阻止H-3-FMLP结合的情况。 FMLP刺激的CD11b上调以及PMN的同型聚集受到LXA的抑制（4）（在10（-9）下，在FMLP剂量 - 响应曲线中给出了1 log单位移动，这是相似的。 LXA（4）r antisera并未改变PMN中FMLP诱导的响应，而是完全阻断了LXA（4）动作。这些结果表明，通过阻塞转录机制或LXA（4）R细胞外域的阻塞来改变LXA（4）R蛋白的表达，从而导致LXA（4）LXA（4）LXA（4）信号传导的特异性结合和阻塞。此外，它们表明，在髓样细胞中，LXA（4）-lxa（4）r相互作用与FMLP的相互作用可以分离，并且LXA（4）（4）在PMN表面调节CD11/18。
Lipoxin A(4) (LXA(4)) interacts with high-affinity receptors in human neutrophils and differentiated HL-60 cells. Recently, we characterized a myeloid-derived cDNA that encodes a LXA(4) high-affinity receptor (LXA(4)R) [Fiore, S., Maddox, J.F., Perez, H.D., and Serhan, C.N. (1994) J. Exp. Med. 180, 253-260] denoted earlier as a related N-formyl peptide receptor (RFP). To examine the selectivity of this receptor we tested its preference for specific binding of H-3-LXA(4) versus H-3-N-formylmethionyl-leucyl-phenylalanine (H-3-FMLP). When receptor-transfected Chinese hamster ovary cells were exposed to either H-3-LXA(4) or H-3-FMLP, the receptor affinity for LXA(4) exceeded by 1000-fold that of FMLP (6.1 nM vs 5 mu M). Upon differentiation, HL-60 cells acquire high-affinity binding sites and respond to both LXA(4) and FMLP. Northern blot analysis of differentiated HL-60 cells using an RFP probe showed a characteristic band at 2.1 kb. Differentiated HL-60 cells exposed to an RFP antisense oligonucleotide selectively lost H-3-LXA(4) binding as well as LXA(4)-stimulated lipid remodeling that paralleled the loss of mRNA for LXA(4)R. In contrast, the specific mRNA for the FMLP receptor, H-3-FMLP specific binding, and FMLP-induced phospholipase D activity were still observed. Treatment of human neutrophils with antisera raised against a peptide in the LXA(4)R third extracellular domain also resulted in selective abrogation of H-3-LXA(4) specific binding with polymorphonuclear leukocytes (PMN) without blocking H-3-FMLP binding. FMLP-stimulated CD11b upregulation as well as homotypic aggregation of PMN was inhibited by LXA(4) (which at 10(-9) M gave similar to 1 log unit shift to the right in the FMLP dose-response curve), The addition of LXA(4)R antisera did not alter FMLP-induced responses in PMN but completely blocked LXA(4) actions. These results indicate that altering the expression of the LXA(4)R protein by blockage of transcriptional mechanisms or hindrance of the LXA(4)R extracellular domains leads to loss of LXA(4) specific binding and blockage of LXA(4) signaling. Moreover, they indicate that in myeloid cells LXA(4)-LXA(4)R interactions are dissociable from those of FMLP and that LXA(4) regulates CD11/18 on the PMN surface.