%0 Journal Article
%T Lipopolysaccharide structure impacts the entry kinetics of bacterial outer membrane vesicles into host cells
%A Anne Marie Krachler
%A Douglas F. Browning
%A Eloise J. O¡¯Donoghue
%A Ewa Bielska
%A Francisco Fernandez-Trillo
%A Luke Alderwick
%A Mohammed Hadis
%A Natalie Sirisaengtaksin
%A Sara Jabbari
%J -
%D 2017
%R 10.1371/journal.ppat.1006760
%X Outer membrane vesicles are nano-sized microvesicles shed from the outer membrane of Gram-negative bacteria and play important roles in immune priming and disease pathogenesis. However, our current mechanistic understanding of vesicle-host cell interactions is limited by a lack of methods to study the rapid kinetics of vesicle entry and cargo delivery to host cells. Here, we describe a highly sensitive method to study the kinetics of vesicle entry into host cells in real-time using a genetically encoded, vesicle-targeted probe. We found that the route of vesicular uptake, and thus entry kinetics and efficiency, are shaped by bacterial cell wall composition. The presence of lipopolysaccharide O antigen enables vesicles to bypass clathrin-mediated endocytosis, which enhances both their entry rate and efficiency into host cells. Collectively, our findings highlight the composition of the bacterial cell wall as a major determinant of secretion-independent delivery of virulence factors during Gram-negative infections
%K Host cells
%K Vesicles
%K Fluorescence resonance energy transfer
%K Endocytosis
%K HeLa cells
%K Antigens
%K Cysteine proteases
%K Bacterial pathogens
%U https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1006760