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首页> 外文期刊>Cell Research >Interaction of non-adherent suspended neutrophils to complement opsonized pathogens: a new assay using optical traps
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Interaction of non-adherent suspended neutrophils to complement opsonized pathogens: a new assay using optical traps

机译：非粘附性悬浮中性粒细胞相互作用以补充调理病原体：使用光阱的新测定

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摘要

Phagocytosis of opsonized pathogens by circulating non-adherent neutrophils is an essential step in host defense, which when overwhelmed contributes to sepsis. To investigate the role played by ligation of complement receptors CR3 and CR4 in non-adherent neutrophils, we designed a novel assay system utilizing dual optical traps, respectively, holding a suspended unactivated cell and presenting a specific ligand-coated bead to the cell surface. We chose anti-CD 18 as an example ligand, mimicking the bacterial opsonizing complement fragment iC3b. Presentation of anti-CD 18-coated beads elicited both pseudopodial protrusion and subsequent phagocytosis. This is in sharp contrast to previously reported responses of adherent neutrophils, which phagocytize opsonized particles without pseudopod formation. We used this same new assay to probe actomyosin pathways in the neutrophil's pseudopodial and phagocytic response. Disruption of actin or inhibition of myosin light-chain kinase dose-dependently reduced pseudopod formation and phagocytosis rates. In summary, ⅰ) the new dual trap assay can be used to study the responses of suspended neutrophils to a variety of ligands, and ⅱ) in a first application of this technique, we found that local ligation of CR3/4 in unactivated neutrophils in suspension induces pseudopod formation and phagocytosis at that site, and that these events occur via an actomyosin-dependent pathway.
机译：循环性非粘附性中性粒细胞吞噬调理病原体的吞噬作用是宿主防御中必不可少的步骤，如果防御能力过强，则会导致败血症。为了研究补体受体CR3和CR4在非粘附性中性粒细胞的连接中所起的作用，我们设计了一种利用双光阱的新型检测系统，分别使用一个未捕获的悬浮细胞和向细胞表面呈递特定配体的珠子。我们选择抗CD 18作为示例配体，模仿细菌调理剂补体片段iC3b。抗CD 18包被的珠的呈现引起假足突和随后的吞噬作用。这与先前报道的粘附性中性粒细胞的反应形成鲜明对比，后者对吞噬调理过的颗粒无伪足形成。我们使用这种相同的新方法来检测中性粒细胞的假足和吞噬反应中的肌动球蛋白途径。肌动蛋白的破坏或肌球蛋白轻链激酶的抑制可剂量依赖性地减少假足的形成和吞噬率。总之，ⅰ）新型双阱测定可用于研究悬浮的中性粒细胞对各种配体的反应，ⅱ）在该技术的首次应用中，我们发现未激活的中性粒细胞中CR3 / 4的局部连接悬浮液在该位点诱导假足形成和吞噬作用，这些事件通过放线菌素依赖性途径发生。

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