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Mulkey, K. M.

Publications and source records attributed to Mulkey, K. M..

2 recordsLinked to original sources

Specificity of Drosophila innubila Nudivirus Infection in Drosophila Cell Culture

Viral host range is an important aspect of both viral biology but also the pragmatic issue of producing viral stocks for experimentation. Host range is important both in terms of the species a virus can infect (taxonomy), and the types of host cells the virus can infect (tropism). Nudiviruses are large DNA viruses that infect several arthropods and are generally poorly studied. Several nudiviruses infect Drosophila species including the Drosophila innubila Nudivirus (DiNV). We aimed to identify cell lines that support the replication of DiNV both for the sake of understanding host range, and to develop a system for large scale production of the virus for infection. We utilized cell lines from the focal host, D. innubila, as well as available cell lines from D. virilis and D. melanogaster and inoculated with both a wild-collected pool of "naive" virus and a D. innubila cell culture-adapted isolate. We found that virus from wild caught flies infected cells from the 3 cell lines and replicate its genome, but the passage 1 fluids from these infections were unable to reinfect upon introduction to new cells. In contrast, a cell culture-adapted strain of DiNV infected the same Drosophila cells (though relatively poorly in D. melanogaster cells) and produced infectious progeny that infected new cells. Thus, our cell culture-adapted virus developed the ability to infect broadly and produce infectious virions.

microbiology↗

The study of the Drosophila innubila Nudivirus in cells and flies

2.Species of the fruit fly, Drosophila, are essential models for investigating host/virus interactions. Research aimed at understanding how hosts mount immune defenses against viruses and how viruses evade those immune defenses relies on host/virus pairs that have evolved together overtime, thus necessitating a need for a DNA virus model system for Drosophila with a virus that naturally infects the host. The Drosophila innubila Nudivirus (DiNV) is an emerging model system poised to take on the role as the DNA virus model for Drosophila. In this paper we describe the development of the DiNV model system with animal and cellular resources. We describe the development of new Drosophila cell lines, virus titration assays, anti-nucleocapsid antibodies, and injection-based DiNV infections in Drosophila innubila. We find that D. innubila cells can be used to grow DiNV for use in further experiments, and that DiNV is highly virulent when injected into adult D. innubila. The resources we have developed enable substantial future research on the DiNV-D. innubila system. 3. Impact statementDNA viruses are the cause of several important and challenging human diseases including hepatitis B, mononucleosis, respiratory disease, gastroenteritis, herpes and chickenpox. Recently there has even been a resurgence of many viral infections due to decreasing rates of vaccination, so to develop better therapeutics and intervention strategies, we must better understand the dynamics of how hosts and DNA viruses interact. The development of a natural DNA virus model in Drosophila creates the opportunity to better understand host/DNA virus interaction. 4. Data summaryAll data and code are available at in our Github repository: available upon publication

microbiology↗