Department of Periodontics, Dental College of Georgia, Augusta University, Augusta, USA.
Department of Oral Biology and Diagnostic Sciences, Augusta University, Augusta, USA.
Sci Rep. 2020 May 4;10(1):7468. doi: 10.1038/s41598-020-64449-8.
Recent epidemiological studies link Periodontal disease(PD) to age-related macular degeneration (AMD). We documented earlier that Porphyromonas gingivalis(Pg), keystone oral-pathobiont, causative of PD, efficiently invades human gingival epithelial and blood-dendritic cells. Here, we investigated the ability of dysbiotic Pg-strains to invade human-retinal pigment epithelial cells(ARPE-19), their survival, intracellular localization, and the pathological effects, as dysfunction of RPEs leads to AMD. We show that live, but not heat-killed Pg-strains adhere to and invade ARPEs. This involves early adhesion to ARPE cell membrane, internalization and localization of Pg within single-membrane vacuoles or cytosol, with some nuclear localization apparent. No degradation of Pg or localization inside double-membrane autophagosomes was evident, with dividing Pg suggesting a metabolically active state during invasion. We found significant downregulation of autophagy-related genes particularly, autophagosome complex. Antibiotic protection-based recovery assay further confirmed distinct processes of adhesion, invasion and amplification of Pg within ARPE cells. This is the first study to demonstrate invasion of human-RPEs, begin to characterize intracellular localization and survival of Pg within these cells. Collectively, invasion of RPE by Pg and its prolonged survival by autophagy evasion within these cells suggest a strong rationale for studying the link between oral infection and AMD pathogenesis in individuals with periodontitis.
最近的流行病学研究将牙周病(PD)与年龄相关性黄斑变性(AMD)联系起来。我们之前已经证明,导致 PD 的关键口腔共生菌牙龈卟啉单胞菌(Pg)能够有效地侵袭人牙龈上皮细胞和血树突状细胞。在这里,我们研究了失调的 Pg 菌株侵袭人视网膜色素上皮细胞(ARPE-19)的能力、它们的存活、细胞内定位以及病理效应,因为 RPE 的功能障碍会导致 AMD。我们表明,活的但不是热杀死的 Pg 菌株能够黏附和侵袭 ARPE。这涉及到早期与 ARPE 细胞膜的黏附、Pg 在单层空泡或细胞质内的内化和定位,并且明显存在一些核定位。没有观察到 Pg 的降解或双层自噬体内部的定位,分裂的 Pg 表明在入侵过程中处于代谢活跃状态。我们发现自噬相关基因,特别是自噬体复合物,显著下调。基于抗生素保护的恢复测定进一步证实了 Pg 在 ARPE 细胞内的黏附、侵袭和扩增的不同过程。这是首次证明人 RPE 被 Pg 侵袭,并开始描述 Pg 在这些细胞内的细胞内定位和存活。总之,Pg 对 RPE 的侵袭以及通过自噬逃避这些细胞内的长期存活表明,在牙周炎患者中研究口腔感染与 AMD 发病机制之间的联系具有很强的合理性。