Analysis of the Langat Virus Genome in Persistent Infection of an Ixodes scapularis Cell Line.
Tick-borne flaviviruses (TBFVs) cause a broad spectrum of disease manifestations ranging from asymptomatic to mild febrile illness and life threatening encephalitis. These single-stranded positive-sense (ss(+)) RNA viruses are naturally maintained in a persistent infection of ixodid ticks and small-medium sized mammals. The development of cell lines from the ixodid ticks has provided a valuable surrogate system for studying the biology of TBFVs in vitro. When we infected ISE6 cells, an Ixodes scapularis embryonic cell line, with Langat virus (LGTV) we observed that the infection proceeded directly into persistence without any cytopathic effect. Analysis of the viral genome at selected time points showed that no defective genomes were generated during LGTV persistence by 10 weeks of cell passage. This was in contrast to LGTV persistence in 293T cells in which defective viral genomes are detectable by five weeks of serial cell passage. We identified two synonymous nucleotide changes i.e., 1893A→C (29% of 5978 reads at 12 h post infection (hpi)) and 2284T→A (34% of 4191 reads at 12 hpi) in the region encoding for the viral protein E. These results suggested that the mechanisms supporting LGTV persistence are different between tick and mammalian cells.
Biology of Vector-Borne Viruses Section, Laboratory of Virology, National Institutes of Health, Hamilton, MT 59840, USA. Luwanika.Mlera@nih.gov.
Luwanika
L
Mlera
Biology of Vector-Borne Viruses Section, Laboratory of Virology, National Institutes of Health, Hamilton, MT 59840, USA. Wessam.Melik@oru.se.
Wessam
W
Melik
Biology of Vector-Borne Viruses Section, Laboratory of Virology, National Institutes of Health, Hamilton, MT 59840, USA. offerdahld@niaid.nih.gov.
Danielle K
DK
Offerdahl
Genomics Unit, Research Technologies Branch, Hamilton, MT 59840, USA. eric.dahlstrom@nih.gov.
Eric
E
Dahlstrom
Genomics Unit, Research Technologies Branch, Hamilton, MT 59840, USA. SPORCELLA@niaid.nih.gov.
Stephen F
SF
Porcella
Biology of Vector-Borne Viruses Section, Laboratory of Virology, National Institutes of Health, Hamilton, MT 59840, USA. mbloom@niaid.nih.gov.
Marshall E
ME
Bloom
D000818:Animals; D002460:Cell Line; D003673:Defective Viruses / Q000235:genetics; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics*; D016679:Genome, Viral; D018884:Ixodes / Q000821:virology*; D008322:Mammals; D017354:Point Mutation; D012692:Serial Passage; D013987:Ticks
ISE6 cells; Ixodes scapularis; Langat virus genome; deep-sequencing; persistent infection; tick-borne flavivirus