Interaction of Neisseria meningitidis with a polarized monolayer of epithelial cells
Interaction of Neisseria meningitidis with a polarized monolayer of epithelial cells
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DOI：
10.1128/iai.65.11.4836-4842.1997
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发表时间：
1997-11-01
影响因子：
3.1
通讯作者：
Nassif, X
中科院分区：
文献类型：
--
作者：
Pujol, C;Eugene, E;Nassif, X
脑膜炎奈瑟菌发病机制中的一个重要步骤是跨越两个细胞屏障，一个在鼻咽，一个在脑。为了探讨这种细菌可以实现这些目标的机制，我们研究了脑膜炎奈瑟菌与生长在过滤单元上的极化紧密连接形成T84细胞的单层之间的相互作用。一种囊化的、有毛的、Opa(-)和OPC(-)的脑膜炎奈瑟氏菌株被证明能够比同基因的无毛的衍生物更有效地附着和跨越这种单层。这种细菌相互作用不影响紧密连接的屏障功能，通过以下评估：(I)跨上皮阻力没有改变，(Ii)没有增加[H-3]菊粉穿透单层，以及(Iii)没有紧密连接蛋白ZO-1的离域。电子显微镜研究和共聚焦检查表明，脑膜炎奈瑟氏菌(I)在附着的细菌下通过肌动蛋白聚合诱导细胞骨架重排，(II)紧密附着在细胞顶膜上，(III)可内化在细胞内。抗毛抗体的免疫荧光染色显示，与这种相互作用的早期阶段相比，在细菌细胞相互作用的后期时间点，脑膜炎双球菌感染显著减少。此外，从受感染的单层中回收的粘附菌有纤毛化、被囊化、Opa(-)和OPC(-)，这是一种与亲本菌株相似的表型。综上所述，这些数据表明，在菌毛介导的黏附之后，脑膜炎奈瑟菌参与了一种亲密附着，这需要一种不同于OpA和Ope的细菌成分，并且脑膜炎球菌通过跨细胞途径而不是旁细胞途径穿过形成紧密连接的上皮细胞的单层。
An important step in the pathogenesis of Neisseria meningitidis is the crossing of two cellular barriers, one in the nasopharynx and one in the brain. To approach the mechanisms by which this bacterium can achieve these goals, we studied the interactions between N. meningitidis and a monolayer of polarized tight junction-forming T84 cells grown on filter units. A capsulated, piliated, Opa(-), and Opc(-) N. meningitidis strain is shown to be capable of adhering to and crossing this monolayer several orders of magnitude more efficiently than an isogenic nonpiliated derivative. This bacterial interaction does not affect the barrier function of tight junctions, as assessed by (i) the absence of modification of the transepithelial resistance, (ii) the lack of increase of [H-3]inulin penetration across the monolayer, and (iii) the absence of delocalization of ZO-1, a tight junction protein. Electron microscopy studies and confocal examinations demonstrated that N. meningitidis (i) induces cytoskeletal rearrangements with actin polymerization beneath adherent bacteria, (ii) is intimately attached to the apical membrane of the cells, and (iii) can be internalized inside cells. Immunofluorescent staining with antipilus antibodies showed evidence that meningococcal piliation was dramatically reduced at later time points of bacterial cell interaction compared to the early phase of this interaction. In addition, adhesive bacteria recovered from an infected monolayer are piliated, capsulated, Opa(-), and Opc(-), a phenotype similar to that of the parental strain. Taken together, these data demonstrate that following pilus-mediated adhesion, N. meningitidis is involved in an intimate attachment which requires a bacterial component different from Opa and Ope and that meningococci cross a monolayer of tight-junction-forming epithelial cells by using a transcellular pathway rather than a paracellular route.