(658d) Virus-Like Proteoliposomes and Proteinaceous Supported Bilayers for Drug Delivery and Screening Applications

Virus-like particles are useful materials for studying virus-host interactions in a safe manner. However, the standard production of pseudovirus based on the Vesicular Stomatitis Virus (VSV) backbone is an intricate procedure that requires trained laboratory personnel. In this presentation, a new strategy for creating virus-like proteoliposomes (VLPLs) and virus-like supported bilayers (VLSBs) is described. This strategy uses a cell blebbing technique to induce the formation of nanoscale vesicles from the plasma membrane of BHK cells expressing the hemagglutinin (HA) fusion protein of influenza X-31. These vesicles and supported bilayers contain HA and are used to carry out single particle membrane fusion events, monitored using total internal reflection fluorescence microscopy. The results of these studies show that the VLPLs and VLSBs contain HA proteins that are fully competent to carry out membrane fusion, including the formation of a fusion pore and the release of fluorophores loaded into vesicles, mimicking the actions of the native virus. This new strategy for creating either spherical or planar geometry virus-like membranes has many potential applications. VLPLs provide an avenue to study fusion proteins of virulent viruses in a safe manner, or they may be used as therapeutic delivery particles to transport beneficial proteins co-expressed in the cells to a target cell. VLSBs can facilitate high throughput screening of antiviral drugs or pathogen-host cell interactions. A few applications will be demonstrated.