Macrophage-derived chemokine induces human eosinophil chemotaxis in a CC chemokine receptor 3- and CC chemokine receptor 4-independent manner
Macrophage-derived chemokine induces human eosinophil chemotaxis in a CC chemokine receptor 3- and CC chemokine receptor 4-independent manner
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DOI：
10.1016/s0091-6749(99)70481-1
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发表时间：
1999-03-01
影响因子：
14.2
通讯作者：
Godiska, R
中科院分区：
文献类型：
--
作者：
Bochner, BS;Bickel, CA;Godiska, R
背景：趋化因子被认为有助于选择性细胞募集，巨噬细胞源性趋化因子 (MDC) 是一种 CC 趋化因子，可引起树突状细胞、单核细胞和活化的自然杀伤细胞的趋化性。 MDC 与 CC 趋化因子受体 4 (CCR4) 结合，但不与 CCR1、CCR2、CCR3、CCR5、CCR6 或 CCR7 结合，目的：我们的目的是通过趋化性和钙流测定来确定 MDC 对人嗜酸性粒细胞的体外活性。方法：从过敏供体的外周血中纯化嗜酸性粒细胞，比较 MDC 与其他 CC 趋化因子的趋化活性在微趋化室测定中。 CCR3 在这些测定中的作用是通过使用 CCR3 阻断抗体来确定的。通过使用 fura-2AM 标记，对用 CCR3 或 CCR4 转染的嗜酸性粒细胞和细胞系进行胞质 Ca++ 动员的测量。采用RT-PCR法测定嗜酸性粒细胞CCR3和CCR4 mRNA的表达。结果：MDC（0.1至100 nmol/L）引起纯化的人嗜酸性粒细胞呈剂量依赖性趋化性（最大与对照的3倍相似）。与其他 CC 趋化因子相比，MDC 和嗜酸性粒细胞趋化因子的嗜酸性粒细胞趋化作用的效力和功效相似，但低于 RANTES、单核细胞趋化蛋白 (MCP)-4 和嗜酸性粒细胞趋化因子-2。 CCR3作为CCR3的阻断单克隆抗体未能抑制MDC诱导的趋化性，此外，用Fura-2AM标记的CCR3转染的人胚胎肾细胞在用eotaxin、eotaxin-2或MCP-B刺激后表现出细胞内游离钙的快速升高，但用MDC刺激后则不然。在 10 ng/mL IL-5 中培养 72 小时的嗜酸性粒细胞在用 eotaxin-2 或 MCP-4 刺激后也表现出细胞内游离钙增加，但用高达 100 nmol/L MDC 则不然。结论：MDC 是嗜酸性粒细胞趋化性的 CCR3 和 CCR4 独立激活剂，但在这些反应过程中它似乎不会引起可测量的胞质钙升高。 MDC 似乎通过 CCR4 之外的另一种受体发挥作用，因此可能有助于嗜酸性粒细胞积累，而不通过 CCR1 至 CCR7 发挥作用。
Background: Chemokines are believed to contribute to selective cell recruitment, Macrophage-derived chemokine (MDC) is a CC chemokine that causes chemotaxis of dendritic cells, monocytes, and activated natural killer cells. MDC binds to CC chemokine receptor 4 (CCR4) but not to CCR1, CCR2, CCR3, CCR5, CCR6, or CCR7,Objective: Our aim was to determine the in vitro activity of MDC on human eosinophils by using chemotaxis and calcium flux assays.Methods: Eosinophils were purified from peripheral blood of allergic donors, and chemotactic activity of MDC and other CC chemokines was compared in microchemotaxis chamber assays. The role of CCR3 in these assays was determined by using a CCR3-bIocking antibody. Measurements of cytosolic Ca++ mobilization were performed by using fura-2AM labeling, with eosinophils and cell lines transfected with CCR3 or CCR4. Eosinophil expression of CCR3 and CCR4 mRNA was determined by using RT-PCR,Results: MDC (0.1 to 100 nmol/L) caused dose-dependent chemotaxis of purified human eosinophils (maximum similar to 3-fold control). Compared with other CC chemokines, the potency and efficacy for eosinophil chemotaxis were similar for MDC and eotaxin but were less than that observed for RANTES, monocyte chemoattractant protein (MCP)-4, and eotaxin-2, Although MDC can act by means of CCR4 RT-PCR analysis failed to reveal CCR4 mRNA in eosinophils, Effects of MDC on eosinophils was also independent of CCR3, as a blocking mAb to CCR3 failed to inhibit MDC-induced chemotaxis, Furthermore, CCR3-transfected human embryonic kidney cells labeled with Fura-2AM exhibited a rapid rise in intracellular free calcium after stimulation with eotaxin, eotaxin-2, or MCP-B but not with MDC. Eosinophils cultured for 72 hours in 10 ng/mL IL-5 also demonstrated increased intracellular free calcium after stimulation with eotaxin-2 or MCP-4, but not with up to 100 nmol/L MDC,Conclusion: MDC is a CCR3- and CCR4-independent activator of eosinophil chemotaxis, but it does not appear to elicit measurable cytosolic calcium elevations during these responses. MDC appears to act by means of another receptor in addition to CCR4 and may therefore contribute to eosinophil accumulation without working through CCR1 to CCR7.