Al-Younes Hesham M, Brinkmann Volker, Meyer Thomas F
Department of Molecular Biology, Max Planck Institute for Infection Biology, D-10117 Berlin, Germany.
Infect Immun. 2004 Aug;72(8):4751-62. doi: 10.1128/IAI.72.8.4751-4762.2004.
Chlamydiae are obligate intracellular pathogens that replicate within a membrane-bound compartment (the inclusion) and are associated with important human diseases, such as trachoma, pneumonia, and atherosclerosis. We have examined the interaction of the host autophagic pathway with Chlamydia trachomatis serovar L2 by using the specific autophagosomal stain monodansylcadaverine, antibodies to autophagosome-associated markers, and traditionally used autophagic inhibitors, particularly 3-methyladenine and amino acids. Chlamydial inclusions did not sequester monodansylcadaverine, suggesting absence of fusion with autophagosomes. Interestingly, exposure of cultures infected for 19 h to 3-methyladenine or single amino acids until the end of infection (44 h) caused various degrees of abnormalities in the inclusion maturation and in the progeny infectivity. Incubation of host cells with chemicals throughout the entire period of infection modulated the growth of Chlamydia even more dramatically. Remarkably, autophagosomal markers MAP-LC3 and calreticulin were redistributed to the inclusion of Chlamydia, a process that appears to be sensitive to 3-methyladenine and some amino acids. The present data indicate the lack of autophagosomal fusion with the inclusion because it was devoid of monodansylcadaverine and no distinct rim of autophagosomal protein-specific staining around the inclusion could be observed. However, high sensitivity of Chlamydia to conditions that could inhibit host autophagic pathway and the close association of MAP-LC3 and calreticulin with the inclusion membrane still suggest a potential role of host autophagy in the pathogenesis of Chlamydia.
衣原体是专性细胞内病原体,在膜结合区室(包涵体)内复制,并与重要的人类疾病相关,如沙眼、肺炎和动脉粥样硬化。我们通过使用特异性自噬体染色单丹磺酰尸胺、自噬体相关标志物抗体以及传统使用的自噬抑制剂,特别是3 - 甲基腺嘌呤和氨基酸,研究了宿主自噬途径与沙眼衣原体血清型L2的相互作用。衣原体包涵体不摄取单丹磺酰尸胺,这表明其与自噬体没有融合。有趣的是,将感染19小时的培养物暴露于3 - 甲基腺嘌呤或单一氨基酸直至感染结束(44小时),导致包涵体成熟和子代感染性出现不同程度的异常。在整个感染期间用化学物质处理宿主细胞对衣原体生长的调节作用更为显著。值得注意的是,自噬体标志物微管相关蛋白轻链3(MAP - LC3)和钙网蛋白重新分布到衣原体包涵体,这一过程似乎对3 - 甲基腺嘌呤和某些氨基酸敏感。目前的数据表明自噬体与包涵体缺乏融合,因为包涵体中没有单丹磺酰尸胺,并且在包涵体周围未观察到自噬体蛋白特异性染色的明显边缘。然而,衣原体对可抑制宿主自噬途径的条件高度敏感,以及MAP - LC3和钙网蛋白与包涵体膜的紧密关联,仍然提示宿主自噬在衣原体发病机制中可能发挥作用。