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首页> 外文期刊>Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society >Lyn-coupled LacCer-enriched lipid rafts are required for CD11b/CD18-mediated neutrophil phagocytosis of nonopsonized microorganisms.
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Lyn-coupled LacCer-enriched lipid rafts are required for CD11b/CD18-mediated neutrophil phagocytosis of nonopsonized microorganisms.

机译：CD11b / CD18介导的非调理微生物的嗜中性白细胞吞噬需要使用Lyn-LacCer富集的脂质筏。

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摘要

The integrin CD11b/CD18 plays a central role in neutrophil phagocytosis. Although CD11b/CD18 binds a wide range of ligands, including C3bi and beta-glucan, and transmits outside-in signaling, the mechanism of this signaling responsible for phagocytosis remains obscure. Here, we report that lactosylceramide (LacCer)-enriched lipid rafts are required for CD11b/CD18-mediated phagocytosis of nonopsonized zymosans (NOZs) by human neutrophils. Anti-CD11b and anti-LacCer antibodies inhibited the binding of NOZs to neutrophils and the phagocytosis of NOZs. During phagocytosis of NOZ, CD11b and LacCer were accumulated and colocalized in the actin-enriched phagocytic cup regions. Immunoprecipitation experiments suggested that CD11b/CD18 was mobilized into the LacCer-enriched lipid rafts during phagocytosis of NOZs. DMSO-treated, neutrophil-like HL-60 cells (D-HL-60 cells) lacking Lyn-coupled, LacCer-mediated signaling showed little phagocytosis of NOZs. However, loading of D-HL-60 cells with C24 fatty acid chain-containing LacCer (C24-LacCer) reconstructed functional Lyn-associated, LacCer-enriched lipid rafts, and restored D-HL-60 cell NOZ phagocytic activity, which was inhibited by anti-LacCer and anti-CD11b antibodies. Lyn knockdown by small interfering RNA blocked the effect of C24:1-LacCer loading on D-HL-60 cell phagocytosis of NOZs. CD11b/CD18 activation experiments indicated phosphorylation of LacCer-associated Lyn by activation of CD11b. Taken together, these observations suggest that CD11b activation causes translocation of CD11b/CD18 into Lyn-coupled, LacCer-enriched lipid rafts, allowing neutrophils to phagocytose NOZs via CD11b/CD18.
机译：整联蛋白CD11b / CD18在嗜中性粒细胞吞噬作用中起重要作用。尽管CD11b / CD18结合了广泛的配体，包括C3bi和β-葡聚糖，并从外而内传递信号，但这种信号的吞噬作用机理仍然不清楚。在这里，我们报告富含乳糖基神经酰胺（LacCer）的脂质筏是人类嗜中性粒细胞CD11b / CD18介导的非调理酵母聚糖（NOZs）吞噬作用所必需的。抗CD11b和抗LacCer抗体抑制NOZ与中性粒细胞的结合以及NOZ的吞噬作用。在NOZ的吞噬过程中，CD11b和LacCer积累并共定位于富含肌动蛋白的吞噬杯区域。免疫沉淀实验表明，CD11b / CD18在NOZ吞噬过程中被转移到富含LacCer的脂质筏中。 DMSO处理的缺乏Lyn偶联LacCer介导的信号转导的中性粒细胞样HL-60细胞（D-HL-60细胞）几乎没有吞噬NOZ。但是，用含有C24脂肪酸链的LacCer（C24-LacCer）装载D-HL-60细胞可重建功能性Lyn相关的，富含LacCer的脂质筏，并恢复了被抑制的D-HL-60细胞NOZ吞噬活性。通过抗LacCer和抗CD11b抗体。小干扰RNA引起的Lyn敲低阻断了C24：1-LacCer负载对NOZs D-HL-60细胞吞噬作用的影响。 CD11b / CD18激活实验表明通过CD11b激活，与LacCer相关的Lyn发生了磷酸化。综上所述，这些观察结果表明CD11b激活导致CD11b / CD18易位到Lyn偶联的富含LacCer的脂质筏中，使中性粒细胞通过CD11b / CD18吞噬NOZ。

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