Cargando…

C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response

Mosquitoes rely on RNA interference (RNAi) as their primary defense against viral infections. To this end, the combination of RNAi and invertebrate cell culture systems has become an invaluable tool in studying virus-vector interactions. Nevertheless, a recent study failed to detect an active RNAi r...

Descripción completa

Detalles Bibliográficos
Autores principales: Brackney, Doug E., Scott, Jaclyn C., Sagawa, Fumihiko, Woodward, Jimmy E., Miller, Neil A., Schilkey, Faye D., Mudge, Joann, Wilusz, Jeffrey, Olson, Ken E., Blair, Carol D., Ebel, Gregory D.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2964293/
https://www.ncbi.nlm.nih.gov/pubmed/21049065
http://dx.doi.org/10.1371/journal.pntd.0000856
_version_ 1782189349272551424
author Brackney, Doug E.
Scott, Jaclyn C.
Sagawa, Fumihiko
Woodward, Jimmy E.
Miller, Neil A.
Schilkey, Faye D.
Mudge, Joann
Wilusz, Jeffrey
Olson, Ken E.
Blair, Carol D.
Ebel, Gregory D.
author_facet Brackney, Doug E.
Scott, Jaclyn C.
Sagawa, Fumihiko
Woodward, Jimmy E.
Miller, Neil A.
Schilkey, Faye D.
Mudge, Joann
Wilusz, Jeffrey
Olson, Ken E.
Blair, Carol D.
Ebel, Gregory D.
author_sort Brackney, Doug E.
collection PubMed
description Mosquitoes rely on RNA interference (RNAi) as their primary defense against viral infections. To this end, the combination of RNAi and invertebrate cell culture systems has become an invaluable tool in studying virus-vector interactions. Nevertheless, a recent study failed to detect an active RNAi response to West Nile virus (WNV) infection in C6/36 (Aedes albopictus) cells, a mosquito cell line frequently used to study arthropod-borne viruses (arboviruses). Therefore, we sought to determine if WNV actively evades the host's RNAi response or if C6/36 cells have a dysfunctional RNAi pathway. C6/36 and Drosophila melanogaster S2 cells were infected with WNV (Flaviviridae), Sindbis virus (SINV, Togaviridae) and La Crosse virus (LACV, Bunyaviridae) and total RNA recovered from cell lysates. Small RNA (sRNA) libraries were constructed and subjected to high-throughput sequencing. In S2 cells, virus-derived small interfering RNAs (viRNAs) from all three viruses were predominantly 21 nt in length, a hallmark of the RNAi pathway. However, in C6/36 cells, viRNAs were primarily 17 nt in length from WNV infected cells and 26–27 nt in length in SINV and LACV infected cells. Furthermore, the origin (positive or negative viral strand) and distribution (position along viral genome) of S2 cell generated viRNA populations was consistent with previously published studies, but the profile of sRNAs isolated from C6/36 cells was altered. In total, these results suggest that C6/36 cells lack a functional antiviral RNAi response. These findings are analogous to the type-I interferon deficiency described in Vero (African green monkey kidney) cells and suggest that C6/36 cells may fail to accurately model mosquito-arbovirus interactions at the molecular level.
format Text
id pubmed-2964293
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-29642932010-11-03 C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response Brackney, Doug E. Scott, Jaclyn C. Sagawa, Fumihiko Woodward, Jimmy E. Miller, Neil A. Schilkey, Faye D. Mudge, Joann Wilusz, Jeffrey Olson, Ken E. Blair, Carol D. Ebel, Gregory D. PLoS Negl Trop Dis Research Article Mosquitoes rely on RNA interference (RNAi) as their primary defense against viral infections. To this end, the combination of RNAi and invertebrate cell culture systems has become an invaluable tool in studying virus-vector interactions. Nevertheless, a recent study failed to detect an active RNAi response to West Nile virus (WNV) infection in C6/36 (Aedes albopictus) cells, a mosquito cell line frequently used to study arthropod-borne viruses (arboviruses). Therefore, we sought to determine if WNV actively evades the host's RNAi response or if C6/36 cells have a dysfunctional RNAi pathway. C6/36 and Drosophila melanogaster S2 cells were infected with WNV (Flaviviridae), Sindbis virus (SINV, Togaviridae) and La Crosse virus (LACV, Bunyaviridae) and total RNA recovered from cell lysates. Small RNA (sRNA) libraries were constructed and subjected to high-throughput sequencing. In S2 cells, virus-derived small interfering RNAs (viRNAs) from all three viruses were predominantly 21 nt in length, a hallmark of the RNAi pathway. However, in C6/36 cells, viRNAs were primarily 17 nt in length from WNV infected cells and 26–27 nt in length in SINV and LACV infected cells. Furthermore, the origin (positive or negative viral strand) and distribution (position along viral genome) of S2 cell generated viRNA populations was consistent with previously published studies, but the profile of sRNAs isolated from C6/36 cells was altered. In total, these results suggest that C6/36 cells lack a functional antiviral RNAi response. These findings are analogous to the type-I interferon deficiency described in Vero (African green monkey kidney) cells and suggest that C6/36 cells may fail to accurately model mosquito-arbovirus interactions at the molecular level. Public Library of Science 2010-10-26 /pmc/articles/PMC2964293/ /pubmed/21049065 http://dx.doi.org/10.1371/journal.pntd.0000856 Text en Brackney et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Brackney, Doug E.
Scott, Jaclyn C.
Sagawa, Fumihiko
Woodward, Jimmy E.
Miller, Neil A.
Schilkey, Faye D.
Mudge, Joann
Wilusz, Jeffrey
Olson, Ken E.
Blair, Carol D.
Ebel, Gregory D.
C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title_full C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title_fullStr C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title_full_unstemmed C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title_short C6/36 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response
title_sort c6/36 aedes albopictus cells have a dysfunctional antiviral rna interference response
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2964293/
https://www.ncbi.nlm.nih.gov/pubmed/21049065
http://dx.doi.org/10.1371/journal.pntd.0000856
work_keys_str_mv AT brackneydouge c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT scottjaclync c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT sagawafumihiko c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT woodwardjimmye c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT millerneila c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT schilkeyfayed c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT mudgejoann c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT wiluszjeffrey c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT olsonkene c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT blaircarold c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse
AT ebelgregoryd c636aedesalbopictuscellshaveadysfunctionalantiviralrnainterferenceresponse