The innate immune system is the first line of defence against viral infections. Conventionally, innate immune activation begins with the detection of foreign nucleic acids by pattern recognition receptors (PRRs), which triggers a signalling cascade that culminates in the production of interferon (IFN) and other inflammatory cytokines and chemokines. Over the past few years, a number of studies have shown that IFN innate immune responses can also be triggered by stressors, such as membrane perturbations, cytoskeletal perturbations, oxidative stress, and endoplasmic reticulum (ER) stress 1–3. One way that some viruses provoke such stress responses is through membrane and cytoskeletal distortions during enveloped virus particle entry. In some cases, the glycoproteins responsible for virus particle entry can also trigger cell-cell fusion. The potential of cell-cell fusion to induce stress-based IFN responses analogous to those triggered by virus-cell fusion has not been addressed until very recently. To investigate if and how cell-cell fusion may induce antiviral mechanisms and IFN responses we used the reptilian reovirus p14 fusion associated small transmembrane (FAST) protein as a model of cell-cell fusion. We found that p14-mediated cell fusion led to the production of low level IFN and upregulation of interferon stimulated genes (ISGs) in a stimulator of interferon genes (STING) and interferon regulatory factor 3 (IRF3) dependent manner. We also observed that multinucleated cells experienced extensive DNA-damage that led to the accumulation of cytosolic DNA in the form of micronuclei. Micronuclei can be detected by cytosolic DNA PRRs like cyclic GMP-AMP synthase (cGAS) and signal IFN production through the cGAS-STING signalling axis. Additionally, early syncytia formation restricted replication of vesicular stomatitis virus (VSV), herpes simplex virus-1 (HSV), and vaccinia virus (VSV) in an IFN and IRF3 independent, and STING dependent manner, suggesting involvement of either a novel antiviral mechanism or suppression of virus replication and spread by biological changes in syncytial cells, such as cell cycle arrest. This study highlights a key role of DNA sensing pathways in the immune response to cell fusion associated stress and points out the importance of fusion kinetics in the selective advantage of syncytial viruses. Understanding the potential of syncytial cells to induce IFN responses and influence viral replication at a mechanistic level is beneficial to the design of improved oncolytic immunotherapy. / Thesis / Master of Science (MSc) / Viruses and their hosts continuously fight each other for survival. The host tries to protect itself from the virus by activating various features of its immune system, while the virus tries to block and evade detection by the immune system. One way that some viruses attempt to bypass the immune system and enhance spread involves expressing proteins that can merge together infected cells with neighboring uninfected cells. Cell-cell fusion disrupts the balanced environment within the cell, which is a form of stress that may activate immune responses. This work investigates if and how host cells may activate the immune system to respond and protect themselves from the cell merging activity of select viruses. We found that the stress associated with existing as a large, fused cell caused DNA damage and fragmentation. These DNA fragments could stimulate key immune sensors and initiate immune responses. We also observed an impaired ability of viruses to infect fused cells, but this restriction was not associated with typical immune responses, suggesting that some other biological change in fused cells created an environment that is not suitable for viral spread. Further investigation is required to fully understand this phenomenon; however, this study highlights some protective mechanisms of the host immune system in response to the stress of viral fusion protein induced cell-cell fusion.
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/26950 |
| Date | January 2021 |
| Creators | Murdza, Tetyana |
| Contributors | Mossman, Karen, Biochemistry and Biomedical Sciences |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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