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...

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Autores principales:	Doug E Brackney, Jaclyn C Scott, Fumihiko Sagawa, Jimmy E Woodward, Neil A Miller, Faye D Schilkey, Joann Mudge, Jeffrey Wilusz, Ken E Olson, Carol D Blair, Gregory D Ebel
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
Acceso en línea:	https://doaj.org/article/9148a471460d481b8aa476f504b8eb70
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spelling	oai:doaj.org-article:9148a471460d481b8aa476f504b8eb702021-11-18T09:13:44ZC6/36 Aedes albopictus cells have a dysfunctional antiviral RNA interference response.1935-27271935-273510.1371/journal.pntd.0000856https://doaj.org/article/9148a471460d481b8aa476f504b8eb702010-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21049065/pdf/?tool=EBIhttps://doaj.org/toc/1935-2727https://doaj.org/toc/1935-2735Mosquitoes 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.Doug E BrackneyJaclyn C ScottFumihiko SagawaJimmy E WoodwardNeil A MillerFaye D SchilkeyJoann MudgeJeffrey WiluszKen E OlsonCarol D BlairGregory D EbelPublic Library of Science (PLoS)articleArctic medicine. Tropical medicineRC955-962Public aspects of medicineRA1-1270ENPLoS Neglected Tropical Diseases, Vol 4, Iss 10, p e856 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
spellingShingle	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Doug E Brackney Jaclyn C Scott Fumihiko Sagawa Jimmy E Woodward Neil A Miller Faye D Schilkey Joann Mudge Jeffrey Wilusz Ken E Olson Carol D Blair Gregory D Ebel C6/36 Aedes albopictus cells have a dysfunctional antiviral RNA interference response.
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	article
author	Doug E Brackney Jaclyn C Scott Fumihiko Sagawa Jimmy E Woodward Neil A Miller Faye D Schilkey Joann Mudge Jeffrey Wilusz Ken E Olson Carol D Blair Gregory D Ebel
author_facet	Doug E Brackney Jaclyn C Scott Fumihiko Sagawa Jimmy E Woodward Neil A Miller Faye D Schilkey Joann Mudge Jeffrey Wilusz Ken E Olson Carol D Blair Gregory D Ebel
author_sort	Doug E Brackney
title	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_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_sort	c6/36 aedes albopictus cells have a dysfunctional antiviral rna interference response.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/9148a471460d481b8aa476f504b8eb70
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C6/36 Aedes albopictus cells have a dysfunctional antiviral RNA interference response.

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