1. Adventitious viruses persistently infect three commonly used mosquito cell lines
- Author
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Claudia Rückert, Michael J. Misencik, Philip M. Armstrong, Nathan D. Grubaugh, James Weger-Lucarelli, Mark D. Stenglein, Gregory D. Ebel, and Doug E. Brackney
- Subjects
0301 basic medicine ,Birnaviridae ,Aedes albopictus ,viruses ,030231 tropical medicine ,Aedes aegypti ,Virus Replication ,Polymerase Chain Reaction ,Virus ,Cell Line ,Flaviviridae ,03 medical and health sciences ,0302 clinical medicine ,Mosquito ,Aedes ,Virology ,Animals ,RNA Viruses ,030304 developmental biology ,RNA, Double-Stranded ,0303 health sciences ,Staining and Labeling ,biology ,Insect-specific viruses ,Sequence Analysis, RNA ,fungi ,High-Throughput Nucleotide Sequencing ,RNA ,RNA virus ,Rhabdoviridae ,biology.organism_classification ,Culex quinquefasciatus ,3. Good health ,Culex ,030104 developmental biology ,Viral replication ,Cell culture ,Bunyaviridae - Abstract
Mosquito cell lines have been used extensively in research to isolate and propagate arthropod-borne viruses and understand virus-vector interactions. Despite their utility as an in vitro tool, these cell lines are poorly defined and may harbor insect-specific viruses. Accordingly, we screened four commonly-used mosquito cell lines, C6/36 and U4.4 cells from Aedes albopictus, Aag2 cells from Aedes aegypti, and Hsu cells from Culex quinquefasciatus, for the presence of adventitious (i.e. exogenous) viruses. All four cell lines stained positive for double-stranded RNA, indicative of RNA virus replication. We subsequently identified viruses infecting Aag2, U4.4 and Hsu cell lines using untargeted next-generation sequencing, but not C6/36 cells. PCR confirmation revealed that these sequences stem from active viral replication and/or integration into the cellular genome. Our results show that these commonly-used mosquito cell lines are persistently-infected with several viruses. This finding may be critical to interpreting data generated in these systems. Centers for Disease Control and Prevention [U50/CCU116806-01, U01/CK000509-01]; US Department of Agriculture Hatch Funds and Multistate Research Project [CONH00773, NE1443]; National Institute of Health, National Institute of Allergy and Infectious Diseases [AI067380, AI099042]; NIH/NCATS [UL1 TR001082] This work was supported in part by grants from the Centers for Disease Control and Prevention (U50/CCU116806-01 and U01/CK000509-01), the US Department of Agriculture Hatch Funds and Multistate Research Project (CONH00773 and NE1443), and the National Institute of Health, National Institute of Allergy and Infectious Diseases (AI067380, AI099042), and NIH/NCATS (UL1 TR001082).
- Published
- 2018