Loading...
Infection specificity and tropism of the Drosophila innubila Nudivirus
Mulkey, Kent Michael
Mulkey, Kent Michael
Citations
Altmetric:
Abstract
The University of Kansas
Viruses affect the health and fitness of all life. Double stranded DNA (dsDNA) viruses such as Herpesviruses and Adenoviruses cause high morbidity and mortality in humans and other animals. However, there are few systems available to study dsDNA viruses in genetically tractable natural hosts. Therefore, we endeavored to develop a natural dsDNA virus model, Drosophila innubila Nudivirus (DiNV), in Drosophila so to study host/dsDNA virus interactions.
Our infection specificity study (Chapter 2) involved determining the replication ability of 2 strains of DiNV in 3 Drosophila cell lines. One of the virus strains tested was derived from wild caught flies and the other was isolated from a culture of primary Drosophila innubila cells that was inoculated with the wildtype virus. We inoculated the 3 cell lines with each of the virus strains and determined whether viral genomic DNA copy number increased as a proxy for virus replication. We found that the naïve virus will infect each of the 3 cell types but does not produce infectious virions that can be passaged. The virus grew best in D. innubila cells, moderately in Drosophila virilis cells and minimally in Drosophila melanogaster cells. Virus adapted to cell culture, however, grew well in both D. innubila and D. virilis cells and minimally in D. melanogaster cells and produced virions that were able to be passaged. Thus, DiNV – especially after adaptation to cells was better able to infect cell lines from species more closely related to its natural host.
We evaluated the virus tropism in Drosophila innubila cells and in male and female adult D. innubila flies (Chapter 3). We observed virus-specific puncta in the cell cytoplasm of D. innubila cells and found similar structures using electron microscopy. We also observed virus particles in the nucleus of infected cells by electron microscopy, but it was rare to see virus particles in the cytoplasm of the cells. We injected the adapted virus into male and female flies and fixed and cut sagittal cryosections of the flies, then stained with an anti-nucleocapsid antibody and then observation by confocal microscopy. We found that DiNV replicates in several tissue types but was most prevalent in gut and reproductive tissues.
None of this work was possible without our development of several assays and techniques to study DiNV. Therefore, Chapter 4 details several of these techniques and our progress with developing cell lines, antibodies, infection and inoculation protocols and viral titering assays. With this work, DiNV is now poised to be a powerful model in the study of host/dsDNA virus interaction.
Description
Date
2025-01-01
Journal Title
Journal ISSN
Volume Title
Publisher
University of Kansas
Collections
Archive Status
This item contains archived web content.
Files
Mulkey_ku_0099D_19928.pdf
Adobe PDF, 40.06 MB
- Embargoed until 2176-05-31
Research Projects
Organizational Units
Journal Issue
Keywords
Molecular biology, Virology, Drosophila innubila, Insect virus, Nudivirus, quantitative PCR, Specificity of infection, Virus tropism