Cooperative vaccinia infection demonstrated at the single-cell level using FluidFM.
The mechanisms used by viruses to enter and replicate within host cells are subjects of intense investigation. These studies are ultimately aimed at development of new drugs that interfere with these processes. Virus entry and infection are generally monitored by dispensing bulk virus suspensions on layers of cells without accounting for the fate of each virion. Here, we take advantage of the recently developed FluidFM to deposit single vaccinia virions onto individual cells in a controlled manner. While the majority of virions were blocked prior to early gene expression, infection of individual cells increased in a nondeterministic fashion with respect to the number of viruses placed. Microscopic analyses of several stages of the virus lifecycle indicated that this was the result of cooperativity between virions during early stages of infection. These findings highlight the importance of performing controlled virus infection experiments at the single cell level.
Institute of Microbiology, Institute of Biomedical Engineering, ETH Zurich, Switzerland.
Philipp
P
Stiefel
Florian I
FI
Schmidt
Pablo
P
Dörig
Pascal
P
Behr
Tomaso
T
Zambelli
Julia A
JA
Vorholt
Jason
J
Mercer
D002462:Cell Membrane / Q000737:chemistry* / Q000276:immunology / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D018625:Microscopy, Atomic Force; D036103:Nanotechnology; D014616:Vaccinia virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology