Inhibition of monocyte chemotaxis to C-C chemokines by antisense oligonucleotide for cytosolic phospholipase A(2)
Inhibition of monocyte chemotaxis to C-C chemokines by antisense oligonucleotide for cytosolic phospholipase A(2)
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DOI：
10.1074/jbc.271.11.6010
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发表时间：
1996-03-15
影响因子：
4.8
通讯作者：
Sozzani, S
中科院分区：
文献类型：
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作者：
Locati, M;Lamorte, G;Sozzani, S
单核细胞趋化蛋白(MCP)-1是趋化因子超家族C-C(或β)分支的成员，在趋化浓度下，可诱导预先标记的人单核细胞快速释放[H-3]花生四烯酸，但不能释放[C-14]油酸。这种作用与胞浆磷脂酶A(2)的磷酸化形式(CPLA(2))对应的免疫反应带的强度增加有关。为了说明CPLA(2)在诱导单核细胞趋化中的作用，用一种特定的反义寡核苷酸处理细胞。经10mU M反义寡核苷酸处理48h后，单核细胞CPLA(2)表达显著降低(57+/-5%；n=4)，MCP-1刺激细胞[H-3]花生四烯酸释放几乎完全抑制(81.8+/-4.2%；n=3)。Cpla(2)反义寡核苷酸对单核细胞对MCP-1的趋化作用也呈浓度依赖性抑制(IC50=1.9+/-1.1 mU；n=3)，在3~10 mU M范围内完全被抑制，对照寡核苷酸对单核细胞的趋化反应无抑制作用。在反义寡核苷酸处理的细胞中，单核细胞对MCP-3、RANTES(调节正常T细胞表达和分泌的活化)和MIP-1α/LD78的迁移也被抑制(70%)。相反，甲酰-甲硫基-亮氨酰苯丙氨酸和C5a这两种“经典”趋化激动剂对趋化反应的影响很小(
Monocyte chemotactic protein (MCP)-1, a member of the C-C (or beta) branch of the chemokine superfamily, at chemotactic concentrations, induced a rapid release of [H-3]arachidonic acid but not of [C-14]oleic acid from prelabeled human monocytes. This effect was associated with an increase in the intensity of the immunoreactive band corresponding to the phosphorylated form of cytosolic phospholipase A(2) (cPLA(2)). To address the role of cPLA(2) in the induction of monocyte chemotaxis, cells were treated with a specific antisense oligonucleotide. Monocytes cultured in the presence of 10 mu M antisense oligonucleotide for 48 h showed a marked decrease (57 +/- 5%; n = 4) of cPLA(2) expression, as evaluated by Western blot analysis and a nearly complete inhibition (81.8 +/- 4.2%; n = 3) of [H-3]arachidonic acid release in MCP-1-stimulated cells. Monocyte chemotaxis in response to MCP-1 also was inhibited in a concentration-dependent manner by cPLA(2) antisense oligonucleotide (IC50 = 1.9 +/- 1.1 mu M; n = 3), with complete inhibition observed between 3 and 10 mu M. No inhibition of chemotactic response was observed in monocytes treated with a control oligonucleotide. Monocyte migration in response to MCP-3, RANTES (regulated on activation normal T cells expressed and secreted), and MIP-1 alpha/LD78 also was inhibited (>70%) in antisense oligonucleotide-treated cells. On the contrary, the chemotactic response elicited by formyl-methionyl-leucyl-phenylalanine and C5a, two ''classical'' chemotactic agonists, was minimally affected (