Transcellular migration of neutrophil granulocytes through the blood-cerebrospinal fluid barrier after infection with Streptococcus suis.
Transcellular migration of neutrophil granulocytes through the blood-cerebrospinal fluid barrier after infection with Streptococcus suis.
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DOI：
10.1186/1742-2094-8-51
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发表时间：
2011-05-18
影响因子：
9.3
通讯作者：
Tenenbaum T
中科院分区：
文献类型：
--
作者：
Wewer C;Seibt A;Wolburg H;Greune L;Schmidt MA;Berger J;Galla HJ;Quitsch U;Schwerk C;Schroten H;Tenenbaum T
细菌性脑膜炎病程中的一个临界点是多形核中性粒细胞(PMN)从血液过度流入大脑。到目前为止，在中枢神经系统疾病中，白细胞通过血脑屏障的旁细胞和跨细胞迁移途径都已被描述。因此，我们研究了炎症条件下PMN通过血-脑脊液屏障迁移的机制。在血-脑脊液屏障的“倒置”Transwell培养模型中，猪链球菌(S.suis)被用来刺激生理相关侧的猪脉络丛上皮细胞(PCPECs)。通过测量TEER和tr-葡聚糖通量来分析屏障功能，并用免疫荧光法观察紧密连接的形态。通过免疫荧光、电子显微镜和流式细胞仪分析，确定PMN迁移的途径和机制。实时定量聚合酶链式反应检测ICAM-1和VCAM-1的表达水平。在这里，我们发现，与其未包裹的突变体相比，经肿瘤坏死因子α刺激或感染猪链球菌10株后，PMN通过PCPECs的迁移显著增加。中性粒细胞迁移对屏障功能的影响不明显，但与猪链球菌感染有关。猪链球菌或肿瘤坏死因子α刺激后，细胞骨架肌动蛋白重排和紧密连接也被观察到。最引人注目的是，PMN优先通过跨细胞途径在PCPEC之间迁移。用Apotome®成像和电子显微镜对PMN迁移过程进行广泛的序列分析表明，细胞旁的PMN迁移恰好在紧密连接之前停止。有趣的是，中性粒细胞随后似乎通过从顶膜发育的漏斗状结构进行跨细胞迁移。值得注意的是，一些PMN在迁移过程中含有细菌。用整合素特异性抗体进行的流式细胞仪和迁移抑制研究表明，PMN的穿透依赖于CD11b/CD18。细胞黏附分子分析显示，经肿瘤坏死因子α和猪链球菌刺激后，细胞间黏附分子-1和血管细胞间黏附分子-1的表达明显增加。我们的数据强调了血-脑脊液屏障作为白细胞进入中枢神经系统的大门的相关性，并提示在细菌性脑膜炎的发病机制中一种新的跨细胞迁移步骤。
A critical point during the course of bacterial meningitis is the excessive influx of polymorphnuclear neutrophils (PMNs) from the blood into the brain. Both paracellular and transcellular routes of leukocyte transmigration through the blood-brain barrier have been described in CNS diseases so far. Thus, we investigated the mechanism of PMN transmigration through the blood-CSF barrier under inflammatory conditions. In an "inverted" Transwell culture model of the blood-CSF barrier, the zoonotic agent Streptococcus suis (S. suis) was used to stimulate porcine choroid plexus epithelial cells (PCPECs) specifically from the physiologically relevant basolateral side. Barrier function was analyzed by measuring TEER and TR-dextran-flux, and tight junction morphology was investigated by immunofluorescence. Route and mechanism of PMN transmigration were determined by immunofluorescence, electron microscopy and FACS analysis. Quantitative real time-PCR was used to determine expression levels of ICAM-1 and VCAM-1. Here, we show that the transmigration of PMNs through PCPECs was significantly higher after stimulation with TNFα or infection with S. suis strain 10 compared to its non-encapsulated mutant. Barrier function was not significantly affected by PMN migration alone, but in combination with S. suis infection. Tight junction and cytoskeletal actin reorganisation were also observed after stimulation with S. suis or TNFα. Most strikingly, PMNs preferentially migrated across PCPECs via the transcellular route. Extensive sequential analyses of the PMN transmigration process with Apotome®-imaging and electron microscopy revealed that paracellular migrating PMNs stop just before tight junctions. Interestingly, PMNs subsequently appeared to proceed by transcellular migration via funnel-like structures developing from the apical membrane. It is noteworthy that some PMNs contained bacteria during the transmigration process. Flow cytometric and transmigration inhibition studies with integrin-specific antibodies showed that PMN traversal is dependent on CD11b/CD18. Analysis of cell adhesion molecules in PCPECs revealed a significant increase of ICAM-1 and VCAM-1 expression after TNFα and S. suis stimulation. Our data underline the relevance of the blood-CSF barrier as a gate for leukocyte entry into the CNS and suggest a novel transcellular migration step during the pathogenesis of bacterial meningitis.
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影响因子：
--
作者：
Joannidis, M;Truebsbach, S;Wiedermann, CJ
通讯作者：
Wiedermann, CJ
影响因子：
4.8
作者：
COLGAN, SP;PARKOS, CA;MADARA, JL
通讯作者：
MADARA, JL
DOI：
10.1093/jnen/59.7.561
发表时间：
2000-07-01
影响因子：
3.2
作者：
Strazielle, N;Ghersi-Egea, JF
通讯作者：
Ghersi-Egea, JF
影响因子：
6.4
作者：
Shahein, YEA;de Andrés, DF;de la Lastra, JMP
通讯作者：
de la Lastra, JMP
影响因子：
4.4
作者：
Edens, HA;Levi, BP;Parkos, CA
通讯作者：
Parkos, CA