RT Journal Article
SR Electronic
T1 Lipopolysaccharide composition determines the preferred route and entry kinetics of bacterial outer membrane vesicles into host cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 084848
DO 10.1101/084848
A1 Eloise J O’Donoghue
A1 Douglas F. Browning
A1 Ewa Bielska
A1 Luke Alderwick
A1 Sara Jabbari
A1 Anne Marie Krachler
YR 2016
UL http://biorxiv.org/content/early/2016/11/01/084848.abstract
AB Outer membrane vesicles are microvesicles shed by 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 kinetics of vesicle entry and cargo delivery to host cells in real-time. Here, we describe a highly sensitive method to study the kinetics of vesicle entry into host cells in real-time using a genetically encoded probe targeted to vesicles. We found that route of vesicular uptake, and thus entry kinetics and efficiency of cargo release, are determined by the chemical composition of the bacterial lipopolysaccharide. The presence of O-antigen facilitates receptor-independent entry, which enhances both rate and efficiency of cargo uptake by host cells. Collectively, our findings highlight the chemical composition of the bacterial cell wall as a major determinant of secretion-independent delivery of virulence factors during Gram-negative infections.