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首页> 外文期刊>Infection and immunity >Surface-Associated Hsp60 Chaperonin of Legionella pneumophila Mediates Invasion in a HeLa Cell Model
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Surface-Associated Hsp60 Chaperonin of Legionella pneumophila Mediates Invasion in a HeLa Cell Model

机译：与表面结合的嗜肺军团菌Hsp60伴侣蛋白介导HeLa细胞模型中的入侵。

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HeLa cells have been previously used to demonstrate that virulent strains of Legionella pneumophila (but not salt-tolerant avirulent strains) efficiently invade nonphagocytic cells. Hsp60, a member of the GroEL family of chaperonins, is displayed on the surface of virulent L. pneumophila (R. A. Gardu?o et al., J. Bacteriol. 180:505–513, 1988). Because Hsp60 is largely involved in protein-protein interactions, we investigated its role in adherence-invasion in the HeLa cell model. Hsp60-specific antibodies inhibited the adherence and invasiveness of two virulent L. pneumophila strains in a dose-dependent manner but had no effect on the association of their salt-tolerant avirulent derivatives with HeLa cells. A monospecific anti-OmpS (major outer membrane protein) serum inhibited the association of both virulent and avirulent strains of L. pneumophila to HeLa cells, suggesting that while both Hsp60 and OmpS may mediate bacterial association to HeLa cells, only virulent strains selectively displayed Hsp60 on their surfaces. Furthermore, the surface-associated Hsp60 of virulent bacterial cells was susceptible to the action of trypsin, which rendered the bacteria noninvasive. Additionally, pretreatment of HeLa cells with purified Hsp60 or precoating of the plastic surface where HeLa cells attached with Hsp60 reduced the adherence and invasiveness of the two virulent strains. Finally, recombinant Hsp60 covalently bound to latex beads promoted the early association of beads with HeLa cells by a factor of 20 over bovine serum albumin (BSA)-coated beads and competed with virulent strains for association with HeLa cells. Hsp60-coated beads were internalized in large numbers by HeLa cells and remained in tight endosomes that did not fuse with other vesicles, whereas internalized BSA-coated beads, for which endocytic trafficking is well established, resided in more loose or elongated endosomes. Mature intracellular forms of L. pneumophila, which were up to 100-fold more efficient than agar-grown bacteria at associating with HeLa cells, were enriched for Hsp60 on the bacterial surface, as determined by immunolocalization techniques. Collectively, these results establish a role for surface-exposed Hsp60 in invasion of HeLa cells by L. pneumophila.
机译：以前曾使用HeLa细胞来证明嗜肺军团菌的强毒株（而不是耐盐的无毒株）有效侵袭非吞噬细胞。 Hsp60是伴侣蛋白GroEL家族的成员，显示在有毒的 L表面。肺炎衣原体（R. A. Gardu？o等人，细菌学杂志（J. Bacteriol。）180：505–513，1988年）。由于Hsp60主要参与蛋白质-蛋白质相互作用，因此我们研究了它在HeLa细胞模型中对黏附侵袭的作用。 Hsp60特异性抗体抑制两种强毒 L的粘附和侵袭性。嗜肺杆菌菌株具有剂量依赖性，但对其耐盐无毒衍生物与HeLa细胞的缔合没有影响。单特异性抗OmpS（主要外膜蛋白）血清可抑制有毒力和无毒力的 L菌株。肺炎对HeLa细胞的影响，这表明虽然Hsp60和OmpS均可介导细菌与HeLa细胞的缔合，但只有强毒株在其表面选择性显示Hsp60。此外，毒性细菌细胞的表面相关的Hsp60对胰蛋白酶的作用敏感，这使细菌无创。另外，用纯化的Hsp60预处理HeLa细胞或在塑料表面进行预涂层（其中HeLa细胞与Hsp60相连）会降低这两种强毒株的粘附和侵袭性。最后，与乳胶珠共价结合的重组Hsp60促进了珠与HeLa细胞的早期缔合，其作用比包被牛血清白蛋白（BSA）的珠高20倍，并与有毒力的菌株竞争与HeLa细胞的缔合。 Hsp60包被的珠子被HeLa细胞大量内在化并保留在不与其他囊泡融合的紧密内体中，而内吞性运输已被很好地建立的内在的BSA包被的珠子则存在于更松散或细长的内体中。 L的成熟细胞内形式。通过免疫定位技术测定，与HeLa细胞结合时，比琼脂生长的细菌效率高100倍的嗜肺菌富集在细菌表面的Hsp60。总的来说，这些结果确立了表面暴露的Hsp60在 L侵袭HeLa细胞中的作用。肺炎。

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