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In situ expression and localization of Neisseria gonorrhoeae opacity proteins in infected epithelial cells: apparent role of Opa proteins in cellular invasion

机译：淋病奈瑟菌不透明蛋白在感染上皮细胞中的原位表达和定位：Opa蛋白在细胞侵袭中的明显作用

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During natural infection, gonococcal opacity proteins (Opa) undergo rapid phase variation, but how this phenomenon contributes to the virulence of the bacteria is not well understood. In the present immunomorphological study we examined the actual Opa status of individual gonococci during various stages of gonococcal infection of Chang epithelial cells, by probing ultrathin sections of infected specimens with Opa-specific monoclonal antibodies. Our results demonstrate a heterogeneous Opa expression during the initial interaction of the bacteria, but an almost 100% expression of one of the probed Opas during their secondary attachment and entry into the host cells, suggesting a role for distinct Opas in cellular penetration. The association between Opa expression, tight attachment, and bacterial invasion into the host cells could be confirmed with isogenic variants that expressed different Opa proteins. Once inside the epithelial cells, both morphologically intact, Opa positive and morphologically disintegrated, Opa negative bacteria were observed. The loss of Opa immunoreactivity in intracellular gonococci could not be related to the presence of a particular Opa protein, but could be mimicked by incubating the organisms with extracts of sonicated uninfected epithelial cells, suggesting that it was caused by host cell proteolytic activity. Taken together, our data suggest that Opa phase transitions confer a functional adaptation of the bacteria enabling host cell penetration.
机译：在自然感染过程中，淋球菌不透明蛋白（Opa）经历了快速的相变，但是这种现象如何导致细菌的毒性尚不清楚。在当前的免疫形态学研究中，我们通过用Opa特异性单克隆抗体探查感染标本的超薄切片，检查了Chang上皮细胞淋球菌感染各个阶段中个体淋球菌的实际Opa状态。我们的结果表明，在细菌的初始相互作用过程中，Opas表达异质，但其中一种被探测的Opas在其次级附着和进入宿主细胞过程中几乎100％的表达，表明不同的Opas在细胞渗透中的作用。 Opa表达，紧密附着和细菌侵入宿主细胞之间的关联可以用表达不同Opa蛋白的等基因变体来证实。一旦进入上皮细胞内部，既观察到形态完整，Opa阳性又发生形态分解，观察到Opa阴性细菌。细胞内淋球菌中Opa免疫反应性的丧失与特定Opa蛋白的存在无关，但可以通过将生物体与超声处理的未感染上皮细胞提取物一起孵育来模拟，这表明它是由宿主细胞的蛋白水解活性引起的。两者合计，我们的数据表明，Opa相变赋予细菌功能适应性，使宿主细胞能够穿透。

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