When: September 30, 2015, 2:00 PM
Location: 3rd Floor Orchard View Room , Discovery Building
Contact: 608-316-4401, email@example.com
Development of tools to study how virus-host cell interactions influence infection spread
Viruses cause a variety of human diseases including AIDS, the common cold, hepatitis, flu and cancer. During a virus infection, the various processes and interactions of the virus, host, and environment form a complex and dynamic system with a heterogeneous range of outcomes. While there has been a strong focus on investigation of molecular mechanisms of virus-host cell interactions within a single-infection cycle and clinical studies that entail multiple infection cycles, relatively less has been done to link how the interactions within a single-infection cycle are reflected over multiple rounds of infection. In this context, the work presented here focuses on the development of experimental and computational quantitative tools to study the factors that influence infection spread from cell to cell in vitro.
In order to investigate this dynamic interaction between virus and host, in the first project we used vesicular stomatitis virus (VSV). We first designed microfluidic channels to study virus spread under stagnant fluid environments. In parallel, to quantify the effects of antiviral responses we developed an antiviral activity assay that measures the overall functional ability of secreted antiviral molecules to inhibit infections. Moreover, using a novel dual color fluorescent reporter system to detect viral replication and cellular antiviral activity; we performed real-time fluorescent microscopy imaging studies of infection spread in conventional macro-culture plates and microchannels. Quantitative analysis of spatial and temporal features of spreading infections in these two different cultures, together with the use of activity assay revealed that the dose and duration dependency of antiviral activation prior to infection ultimately determined the spread or arrest of the infection. For the second project, using the image processing and analysis tools, we focused on rhinovirus (RV), which contributes to a variety of respiratory illnesses. Aiming to link the genetic diversity observed in different RV strains with differences in the severity of infections observed clinically, we developed single-cell image analysis workflows. The workflows enabled tracking of individual cells and extraction of fluorescent intensities from different cellular compartments in a dual color fluorescent transfection assay. Comparison of different strains revealed the differential disruption of nucleo-cytoplasmic transport, a key mechanism for intracellular antiviral signaling. Extending these studies to probe spatial and temporal dynamics of spreading RV infections, we quantified the spread patterns of a recombinant fluorescent RV strain and extracted parameters that can be further used for characterizing the various features of the spread phenotypes of different RV strains. The multi-cycle infection analysis of various RV strains may help gain insights into varying degrees of illness severities observed for infections of different RV strains. Overall, this work demonstrates the potential of these new experimental and computational tools for enabling a more quantitative approach to elucidating the dynamic and complex mechanisms of virus-host interactions.
All SysBioM (Systems Biology in Madison) sponsored talks take place on Wednesdays at 2 p.m in the 3rd floor Orchard View room of the Discovery Building. Talks are open to the public. Access to the room is via the elevator behind Aldo’s Cafe in the Northeast corner of the building.