Abstract
The study of pathogen interactions with eukaryotic host cells requires the introduction of fluorescent probes to visualize processes such as endocytosis, intracellular transport or host cell manipulation by the pathogen. Here, three types of fluorescent nanoparticles (NPs), i.e. Rhodamine-labeled polymethacrylate (PMA) NPs, silica NPs and gold NPs, were employed to label the host cellular endolysosomal system and monitor manipulations by the pathogen Salmonella enterica. Using live cell imaging, we investigated the performance of NPs in cellular uptake, labeling of endocytic vesicles and lysosomes, as well as interaction with the pathogen. We show that fluorescent gold and silica, but not PMA NPs appropriately label host cell structures and efficiently track rearrangements of the host endosomal system by the activities of intracellular Salmonella. Silica NPs slightly aggregated and located in Salmonella-induced compartments as isolated dots, while gold NPs distributed uniformly inside such structures. Both silica and gold NPs exhibited no adverse impact on either host cells or pathogens, and are versatile tools for infection biology.
Highlights
The understanding of microbial virulence functions involves the analyses of interactions between pathogens and host cells
We describe the systematic analysis of a variety of uorescent NPs for applications as tracers in cellular microbiology and their performance in the S. typhimurium infection model
Uorescent polymethacrylate NPs (PMA NPs), silica NPs and gold NPs were applied for labeling of the endosomal system and monitoring the arrangement of endolysosomal vesicles by the intracellular pathogen S. typhimurium
Summary
The understanding of microbial virulence functions involves the analyses of interactions between pathogens and host cells. To label the incoming endosomal vesicles and observe their fusion with the SCV and SIF, HeLa cells were rst infected with S. typhimurium as described before, at 1 h p.i. NPs were added to incubate with the cells.
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