A microfluidic-based genetic screen to identify microbial virulence factors that inhibit dendritic cell migration

Microbial pathogens are able to modulate host cells and evade the immune system by multiple mechanisms. For example, Salmonella injects effector proteins into host cells and evades the host immune system in part by inhibiting dendritic cell (DC) migration. The identification of microbial factors tha...

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Bibliographic Details
Published in:Integrative biology (Cambridge) 2014-04, Vol.6 (4), p.438-449
Main Authors: McLaughlin, Laura M, Xu, Hui, Carden, Sarah E, Fisher, Samantha, Reyes, Monique, Heilshorn, Sarah C, Monack, Denise M
Format: Article
Language:English
Subjects:
Animals
Cell Movement - immunology
Chemokine CCL19 - immunology
Dendritic Cells - immunology
Dendritic Cells - microbiology
Host-Pathogen Interactions
Mice
Mice, 129 Strain
Microfluidics
Microscopy, Fluorescence
Microscopy, Phase-Contrast
Salmonella Infections - immunology
Salmonella typhimurium - genetics
Salmonella typhimurium - immunology
Salmonidae
Sequence Deletion - immunology
Statistics, Nonparametric
Virulence Factors - genetics
Virulence Factors - immunology
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Summary:Microbial pathogens are able to modulate host cells and evade the immune system by multiple mechanisms. For example, Salmonella injects effector proteins into host cells and evades the host immune system in part by inhibiting dendritic cell (DC) migration. The identification of microbial factors that modulate normal host functions should lead to the development of new classes of therapeutics that target these pathways. Current screening methods to identify either host or pathogen genes involved in modulating migration towards a chemical signal are limited because they do not employ stable, precisely controlled chemical gradients. Here, we develop a positive selection microfluidic-based genetic screen that allows us to identify Salmonella virulence factors that manipulate DC migration within stable, linear chemokine gradients. Our screen identified 7 Salmonella effectors (SseF, SifA, SspH2, SlrP, PipB2, SpiC and SseI) that inhibit DC chemotaxis toward CCL19. This method is widely applicable for identifying novel microbial factors that influence normal host cell chemotaxis as well as revealing new mammalian genes involved in directed cell migration.
ISSN:1757-9694
1757-9708
DOI:10.1039/c3ib40177d