Nolan Sabrina J, Romano Julia D, Coppens Isabelle
Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
PLoS Pathog. 2017 Jun 1;13(6):e1006362. doi: 10.1371/journal.ppat.1006362. eCollection 2017 Jun.
Toxoplasma is an obligate intracellular parasite that replicates in mammalian cells within a parasitophorous vacuole (PV) that does not fuse with any host organelles. One mechanism developed by the parasite for nutrient acquisition is the attraction of host organelles to the PV. Here, we examined the exploitation of host lipid droplets (LD), ubiquitous fat storage organelles, by Toxoplasma. We show that Toxoplasma replication is reduced in host cells that are depleted of LD, or impaired in TAG lipolysis or fatty acid catabolism. In infected cells, the number of host LD and the expression of host LD-associated genes (ADRP, DGAT2), progressively increase until the onset of parasite replication. Throughout infection, the PV are surrounded by host LD. Toxoplasma is capable of accessing lipids stored in host LD and incorporates these lipids into its own membranes and LD. Exogenous addition of oleic acid stimulates LD biogenesis in the host cell and results in the overaccumulation of neutral lipids in very large LD inside the parasite. To access LD-derived lipids, Toxoplasma intercepts and internalizes within the PV host LD, some of which remaining associated with Rab7, which become wrapped by an intravacuolar network of membranes (IVN). Mutant parasites impaired in IVN formation display diminished capacity of lipid uptake from host LD. Moreover, parasites lacking an IVN-localized phospholipase A2 are less proficient in salvaging lipids from host LD in the PV, suggesting a major contribution of the IVN for host LD processing in the PV and, thus lipid content release. Interestingly, gavage of parasites with lipids unveils, for the first time, the presence in Toxoplasma of endocytic-like structures containing lipidic material originating from the PV lumen. This study highlights the reliance of Toxoplasma on host LD for its intracellular development and the parasite's capability in scavenging neutral lipids from host LD.
弓形虫是一种专性细胞内寄生虫,在一个不与任何宿主细胞器融合的寄生泡(PV)内的哺乳动物细胞中进行复制。寄生虫为获取营养而发展出的一种机制是将宿主细胞器吸引到寄生泡。在此,我们研究了弓形虫对宿主脂滴(LD)的利用情况,脂滴是普遍存在的脂肪储存细胞器。我们发现,在缺乏脂滴、三酰甘油脂解或脂肪酸分解代谢受损的宿主细胞中,弓形虫的复制会减少。在受感染的细胞中,宿主脂滴的数量以及与宿主脂滴相关基因(ADRP、DGAT2)的表达会逐渐增加,直至寄生虫开始复制。在整个感染过程中,寄生泡都被宿主脂滴包围。弓形虫能够获取宿主脂滴中储存的脂质,并将这些脂质整合到自身的膜和脂滴中。外源添加油酸会刺激宿主细胞中的脂滴生物合成,并导致寄生虫内非常大的脂滴中中性脂质过度积累。为了获取源自脂滴的脂质,弓形虫在寄生泡内拦截并内化宿主脂滴,其中一些脂滴仍与Rab7相关联,随后被泡内膜网络(IVN)包裹。在IVN形成方面受损的突变寄生虫从宿主脂滴摄取脂质的能力减弱。此外,缺乏定位于IVN的磷脂酶A2的寄生虫在从寄生泡中的宿主脂滴中挽救脂质方面效率较低,这表明IVN对寄生泡中宿主脂滴的处理以及脂质含量释放起主要作用。有趣的是,用脂质灌喂寄生虫首次揭示了弓形虫中存在含有源自寄生泡腔脂质物质的内吞样结构。这项研究突出了弓形虫在细胞内发育过程中对宿主脂滴的依赖性以及寄生虫从宿主脂滴中清除中性脂质的能力。