scholarly journals Adventitious viruses persistently infect three commonly used mosquito cell lines

2018 ◽  
Author(s):  
James Weger-Lucarelli ◽  
Claudia Rückert ◽  
Nathan D. Grubaugh ◽  
Michael J. Misencik ◽  
Philip M. Armstrong ◽  
...  
Keyword(s):  
Cell Lines ◽  
Rna Virus ◽  
Mosquito Cell ◽  
Double Stranded Rna ◽  
Active Viral Replication ◽  
Mosquito Cell Lines ◽  
Persistently Infect ◽  
Generation Sequencing ◽  
Cellular Genome

AbstractMosquito cell lines were first established in the 1960’s and have been used extensively in research to isolate and propagate arthropod-borne (arbo-) viruses, study the invertebrate immune system, and understand virus-vector interactions. Despite their utility as anin vitrotool, these cell lines are poorly defined and may harbor insect-specific viruses that could impact experimental results. Accordingly, we screened four commonly-used mosquito cell lines, C6/36 and U4.4 cells fromAedes albopictus, Aag2 cells fromAedes aegypti, and Hsu cells fromCulex quinquefasciatus, for the presence of adventitious viruses. All four cell lines stained positive for double-stranded RNA by immunofluorescence, 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. Sequences from viruses in the familiesBirnaviridae,Bunyaviridae, Flaviviridae,andRhabdoviridaewere abundant in the mosquito cell lines. 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.

scholarly journals Adventitious viruses persistently infect three commonly used mosquito cell lines

Virology ◽  
2018 ◽  
Vol 521 ◽  
pp. 175-180 ◽  
Author(s):  
James Weger-Lucarelli ◽  
Claudia Rückert ◽  
Nathan D. Grubaugh ◽  
Michael J. Misencik ◽  
Philip M. Armstrong ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mosquito Cell ◽  
Mosquito Cell Lines ◽  
Persistently Infect

scholarly journals Aedes aegypti (Aag2)-derived clonal mosquito cell lines reveal the effects of pre-existing persistent infection with the insect-specific bunyavirus Phasi Charoen-like virus on arbovirus replication

2019 ◽  
Vol 13 (11) ◽  
pp. e0007346 ◽  
Author(s):  
Anthony C. Fredericks ◽  
Tiffany A. Russell ◽  
Louisa E. Wallace ◽  
Andrew D. Davidson ◽  
Ana Fernandez-Sesma ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Cell Lines ◽  
Persistent Infection ◽  
Mosquito Cell ◽  
Mosquito Cell Lines

A phase I study of reolysin given intravenously to patients with advanced malignancies

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 3064-3064 ◽  
Author(s):  
L. Vidal ◽  
H. Pandha ◽  
J. Spicer ◽  
K. J. Harrington ◽  
S. Allen ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Viral Replication ◽  
Tumor Necrosis ◽  
Rna Virus ◽  
Maximum Tolerated Dose ◽  
Systemic Delivery ◽  
Double Stranded Rna ◽  
Infectious Dose ◽  
Viral Shedding

3064 Background: Reovirus is a double-stranded RNA virus with minimal pathogenicity in humans that selectively replicates in cells with activated Ras. Activated Ras inhibits the anti-viral effects of double stranded RNA-activated protein kinase (PKR). Reovirus serotype 3 Dearing has selective antitumor activity, in vitro and in tumour xenograft models. Methods: Reolysin was administered as a 1-hr IV infusion every 4-weeks initially for one day; then 3 days then 5-days every 4 weeks. The starting dose was 1×108 tissue culture infectious dose (TCID50) increasing in successive cohorts until observation of drug-related toxicity ≥ grade 2. Endpoints were safety, viral replication, viral shedding, evaluation of immune response and antitumor activity. Results: 24 patients (pts) (median age 60; ECOG 1, 16 males) have been entered into the first 7 cohorts at the following dose levels: 1×108 for 1-day, 1×108 for 3-days and 1×108, 3×108, 1×109, 3×109 and 1×1010 TCID50 for 5-days. A maximum tolerated dose (MTD) has not been reached and no dose-limiting toxicities have been observed. Toxicities have been mild (grade 1 or 2) and have included chills, fever, headache, runny nose, fatigue and myelosuppression. Reverse transcription polymerase chain reaction (RT-PCR) studies of blood, urine, stool and sputum post reovirus administration were negative for viral shedding for all treated pts. All but one pt had neutralising anti-reovirus antibodies detectable pre-treatment. Titres increased after 1-week of treatment and remained high during subsequent courses of treatment. Two pts with metastatic colorectal cancer treated at 3×108 and 3×109 TCID50 had CEA tumour marker reduction by 60% and 27% receiving 6 and 3 courses of treatment respectively. One pt with metastatic pancreatic cancer received 4 courses of treatment with stable disease. One pt with metastatic prostate cancer had a 50% decrease in PSA after treatment at 3×109 TCID50, with evidence of tumor necrosis on CT scanning. Intratumoral reovirus replication has been detected by electron microscopy in tumur biopsies. Conclusions: Reolysin is well tolerated with minimal toxicity. No viral shedding has been detected. Virus-induced tumor necrosis associated with intratumor viral replication after systemic delivery has been observed. [Table: see text]

Stable transformation of mosquito cell lines using a hsp70::neo fusion gene

Gene ◽  
1993 ◽  
Vol 136 (1-2) ◽  
pp. 129-136 ◽  
Author(s):  
Gareth J. Lycett ◽  
Julian M. Crampton
Keyword(s):  
Cell Lines ◽  
Fusion Gene ◽  
Stable Transformation ◽  
Mosquito Cell ◽  
Mosquito Cell Lines

scholarly journals His-154 is involved in the linkage of the Saccharomyces cerevisiae L-A double-stranded RNA virus Gag protein to the cap structure of mRNAs and is essential for M1 satellite virus expression.

1994 ◽  
Vol 14 (4) ◽  
pp. 2664-2674 ◽  
Author(s):  
A Blanc ◽  
J C Ribas ◽  
R B Wickner ◽  
N Sonenberg
Keyword(s):  
Rna Virus ◽  
Specific Binding ◽  
Covalent Bond ◽  
Gag Protein ◽  
Double Stranded Rna ◽  
Satellite Virus ◽  
Eukaryotic Mrnas ◽  
Virus Expression

The coat protein (Gag) of the double-stranded RNA virus L-A was previously shown to form a covalent bond with the cap structure of eukaryotic mRNAs. Here, we identify the linkage as a phosphoroimidazole bond between the alpha phosphate of the cap structure and a nitrogen in the Gag protein His-154 imidazole side chain. Mutations of His-154 abrogate the ability of Gag to bind to the cap structure, without affecting cap recognition, in vivo virus particle formation from an L-A cDNA clone, or in vitro specific binding and replication of plus-stranded single-stranded RNA. However, genetic analyses demonstrate that His-154 is essential for M1 satellite virus expression.

scholarly journals In vitro selection of packaging sites in a double-stranded RNA virus.

1997 ◽  
Vol 71 (3) ◽  
pp. 2157-2162 ◽  
Author(s):  
W Yao ◽  
K Adelman ◽  
J A Bruenn
Keyword(s):  
In Vitro Selection ◽  
Rna Virus ◽  
Double Stranded Rna ◽  
Selection Of
Sign in / Sign up

Export Citation Format

Share Document