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Ichnovirus infection of an established gypsy moth cell line
1Department of Microbiology & Immunology, Dalhousie University, Halifax, Nova Scotia, Canada.
Abstract:
In the present study, a lepidopteran cell line (Ld-652Y, from the gypsy moth, Lymantria dispar) exposed to Hyposoter fugitivus polydnavirus (HfPV) was found to display a variety of cytopathic effects. These included a transient inhibition of cell proliferation, rounding up, aggregation and apoptosis. In addition, unusual paracrystalline structures appeared within the lumen of the rough endoplasmic reticulum; similar structures were observed in the spherulocytes of parasitized Malacosoma disstria. Following Coomassie Blue staining, two new cell-associated polypeptides were detected; one of these, an 8 kDa polypeptide, could also be observed following exposure of LD-652Y cells to media taken from infected cultures or to cell-free haemolymph from parasitized M. disstria. After a period of 2-4 weeks, the L. dispar cell cultures were observed to largely recover from the effects of exposure to virus, and resumed proliferation; "transformed' cell populations tended to form aggregates, and adhered less tightly to the substrate. Viral DNA was stably maintained in all recovered cell lines, possibly in chromosomally integrated form.
Insights
Gypsy moth cells infected with Hyposoter fugitivus polydnavirus (HfPV) showed cell damage and new protein production. Recovered cells maintained viral DNA, suggesting integration.
Area of Science:
- Entomology
- Molecular Biology
- Cell Biology
Background:
- Polydnaviruses are essential symbionts of parasitic wasps, playing a crucial role in host immune suppression and development.
- Lepidopteran cell lines provide valuable models for studying insect-specific viruses and their cellular interactions.
- Understanding virus-cell interactions is key to insect pest management and biological control strategies.
Purpose of the Study:
- To investigate the cytopathic effects of Hyposoter fugitivus polydnavirus (HfPV) on a Lymantria dispar (gypsy moth) cell line (Ld-652Y).
- To identify and characterize cellular and molecular changes induced by HfPV infection.
- To examine the recovery and long-term fate of HfPV-infected insect cells.
Main Methods:
- Exposure of Ld-652Y cells to HfPV and observation of cytopathic effects.
- Microscopic analysis to identify structural changes within cells.
- Coomassie Blue staining and polypeptide analysis to detect changes in protein expression.
- Monitoring of cell proliferation, aggregation, and substrate adhesion.
- Detection of viral DNA in recovered cell lines.
Main Results:
- HfPV infection caused transient inhibition of cell proliferation, cell rounding, aggregation, and apoptosis in Ld-652Y cells.
- Unusual paracrystalline structures were observed in the rough endoplasmic reticulum of infected cells and in parasitized M. disstria spherulocytes.
- Two new cell-associated polypeptides were detected, including an 8 kDa polypeptide found in cells exposed to infected media or hemolymph.
- Recovered Ld-652Y cell cultures resumed proliferation after 2-4 weeks, exhibiting altered aggregation and adhesion properties.
- Viral DNA was stably maintained in recovered cell lines, with evidence suggesting chromosomal integration.
Conclusions:
- HfPV induces significant cytopathic effects and alterations in protein expression in Ld-652Y cells.
- The observed paracrystalline structures and protein changes highlight novel virus-host interactions.
- Lepidopteran cells can recover from HfPV infection, with viral DNA persistence potentially mediated by integration.
- This study provides insights into polydnavirus pathogenesis and host cell response in insects.