| A new method was developed to quantitatively characterize the dynamics of virus-host interaction in a parallel, efficient manner. This experimental system enabled the detection of viral proteins as well as host cell components as a function of time by indirect immunofluorescence. The viral propagation velocity was used to quantify viral spread, and this quantity provided an integrated measure of the various reactions and transport processes required for successful viral infections in vitro.;Microcontact printing was used to generate arrays with various geometries of patterned self-assembled monolayers of alkanethiols on gold surfaces. These patterned surfaces served as substrates for the culture of baby hamster kidney (BHK-21) and murine astrocytoma (DBT) cells, which were used as example systems for studying the propagation of vesicular stomatitis virus (VSV), a model RNA virus.;Different temporal stages of viral infection were monitored and resolved in a parallel manner by using micropattemed host cells. These stages included initiation of infection, based on G protein expression; cell-cell fusion, based on virus-induced clustering of cell nuclei; cytoskeletal disorganization, based on localized release of cells from the surface and disintegration of actin filaments; and cellular antiviral responses, monitored by nitric oxide synthase induction and expression. Multiple environmental factors influenced VSV propagation and antiviral responses, including cell type, the presence of adsorbed proteins, and surface chemistry. In general, the presence of adsorbed protein reduced viral spread in vitro, and DBT cells, but not BHK cells, were able to contain viral spread if sufficient antiviral activity was initiated early in the infection process.;Additionally, the long-term dynamics of cells on micropatterned surfaces were quantified by characterizing the pattern fidelity, or the adherence to and maintenance of the underlying chemical pattern by cells. Again, surface chemistry, cell type, presence of preadsorbed protein, and pattern geometry all affect the long-term behavior of populations of micropatterned cells, with protein-coated surfaces generally resulting in the least stable cellular patterns over long periods of time.;This work sets a foundation for parallel, high-throughput characterization of viral and cellular processes and the successful incorporation of micropatterned cell cultures into an increasing variety of cell-based assays. |
