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首页> 外文期刊>Journal of neuroinflammation >Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro
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Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro

机译：细菌感染后多形核中性粒细胞和单核细胞通过人血脑脊液屏障的迁移

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Background Bacterial invasion through the blood-cerebrospinal fluid barrier (BCSFB) during bacterial meningitis causes secretion of proinflammatory cytokines/chemokines followed by the recruitment of leukocytes into the CNS. In this study, we analyzed the cellular and molecular mechanisms of polymorphonuclear neutrophil (PMN) and monocyte transepithelial transmigration (TM) across the BCSFB after bacterial infection. Methods Using an inverted transwell filter system of human choroid plexus papilloma cells (HIBCPP), we studied leukocyte TM rates, the migration route by immunofluorescence, transmission electron microscopy and focused ion beam/scanning electron microscopy, the secretion of cytokines/chemokines by cytokine bead array and posttranslational modification of the signal regulatory protein (SIRP) α via western blot. Results PMNs showed a significantly increased TM across HIBCPP after infection with wild-type Neisseria meningitidis (MC58). In contrast, a significantly decreased monocyte transmigration rate after bacterial infection of HIBCPP could be observed. Interestingly, in co-culture experiments with PMNs and monocytes, TM of monocytes was significantly enhanced. Analysis of paracellular permeability and transepithelial electrical resistance confirmed an intact barrier function during leukocyte TM. With the help of the different imaging techniques we could provide evidence for para- as well as for transcellular migrating leukocytes. Further analysis of secreted cytokines/chemokines showed a distinct pattern after stimulation and transmigration of PMNs and monocytes. Moreover, the transmembrane glycoprotein SIRPα was deglycosylated in monocytes, but not in PMNs, after bacterial infection. Conclusions Our findings demonstrate that PMNs and monoctyes differentially migrate in a human BCSFB model after bacterial infection. Cytokines and chemokines as well as transmembrane proteins such as SIRPα may be involved in this process.
机译：背景技术细菌性脑膜炎期间细菌通过血脑脊液屏障（BCSFB）的入侵导致促炎性细胞因子/趋化因子的分泌，随后将白细胞募集到CNS中。在这项研究中，我们分析了细菌感染后多形核中性粒细胞（PMN）和单核细胞跨上皮细胞迁移（TM）跨BCSFB的细胞和分子机制。方法利用人脉络丛乳头状瘤细胞（HIBCPP）的倒置超滤系统，研究白细胞TM率，免疫荧光的迁移途径，透射电镜和聚焦离子束/扫描电镜，细胞因子珠的细胞因子/趋化因子分泌。通过蛋白质印迹对信号调节蛋白（SIRP）α进行阵列和翻译后修饰。结果野生型脑膜炎奈瑟菌（MC58）感染后，PMNs在整个HIBCPP中显示TM显着增加。相比之下，HIBCPP细菌感染后，单核细胞迁移率显着降低。有趣的是，在与PMN和单核细胞的共培养实验中，单核细胞的TM显着提高。对细胞旁通透性和跨上皮电阻的分析证实了白细胞TM期间完整的屏障功能。借助不同的成像技术，我们可以为跨细胞以及跨细胞迁移的白细胞提供证据。对PMN和单核细胞的刺激和迁移后，对分泌的细胞因子/趋化因子的进一步分析显示出不同的模式。此外，细菌感染后，跨膜糖蛋白SIRPα在单核细胞中被去糖基化，而在PMN中则没有。结论我们的发现表明，在细菌感染后，PMN和单核细胞在人BCSFB模型中差异迁移。细胞因子和趋化因子以及跨膜蛋白（例如SIRPα）可能参与此过程。

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