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; Schilkey, Faye; Mudge, Joann; Wilusz, Jeffrey; Olson, Ken E.; Blair, Carol D.; Ebel, Gregory D.

doi:10.1371/journal.pntd.0000856

Cited by 286 publications

(294 citation statements)

References 49 publications

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“…21 Kenney and others 43 reported that Nahuirim virus, a newly characterized flavivirus from Brazil, reduces replication of WNV, Japanese encephalitis, and St. Louis encephalitis viruses in dually infected mosquito cell cultures. All of these in vitro experiments were conducted using C6/36 cells, which, unlike live mosquitoes, do not have a functional antiviral RNAi response, 44 thus the biological relevance of these in vitro results is uncertain.…”

Section: Discussionmentioning

confidence: 99%

Insect-Specific Viruses Detected in Laboratory Mosquito Colonies and Their Potential Implications for Experiments Evaluating Arbovirus Vector Competence

Bolling¹,

Vasilakis²,

Guzmán³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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Abstract. Recently, there has been a dramatic increase in the detection and characterization of insect-specific viruses in field-collected mosquitoes. Evidence suggests that these viruses are ubiquitous in nature and that many are maintained by vertical transmission in mosquito populations. Some studies suggest that the presence of insect-specific viruses may inhibit replication of a super-infecting arbovirus, thus altering vector competence of the mosquito host. Accordingly, we screened our laboratory mosquito colonies for insect-specific viruses. Pools of colony mosquitoes were homogenized and inoculated into cultures of Aedes albopictus (C6/36) cells. The infected cells were examined by electron microscopy and deep sequencing was performed on RNA extracts. Electron micrograph images indicated the presence of three different viruses in three of our laboratory mosquito colonies. Potential implications of these findings for vector competence studies are discussed.

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Section: Discussionmentioning

confidence: 99%

Insect-Specific Viruses Detected in Laboratory Mosquito Colonies and Their Potential Implications for Experiments Evaluating Arbovirus Vector Competence

Bolling¹,

Vasilakis²,

Guzmán³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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“…Since then, work on the subject has centered exclusively on mosquito-arbovirus experimental systems. In Aedes mosquitoes and cell lines, an expanded family of Piwi proteins is expressed in somatic tissues, and viral-derived piRNAs (vpiRNAs) are produced from the genomes of several arboviruses (23,(25)(26)(27)(28)(29). Functional links among the piRNA pathway, arbovirus replication, and vpiRNA production have been described as well.…”

Section: Significancementioning

confidence: 99%

piRNA pathway is not required for antiviral defense in Drosophila melanogaster

Petit

Mongelli

Frangeul

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Since its discovery, RNA interference has been identified as involved in many different cellular processes, and as a natural antiviral response in plants, nematodes, and insects. In insects, the small interfering RNA (siRNA) pathway is the major antiviral response. In recent years, the Piwi-interacting RNA (piRNA) pathway also has been implicated in antiviral defense in mosquitoes infected with arboviruses. Using Drosophila melanogaster and an array of viruses that infect the fruit fly acutely or persistently or are vertically transmitted through the germ line, we investigated in detail the extent to which the piRNA pathway contributes to antiviral defense in adult flies. Following virus infection, the survival and viral titers of Piwi, Aubergine, Argonaute-3, and Zucchini mutant flies were similar to those of wild type flies. Using next-generation sequencing of small RNAs from wild type and siRNA mutant flies, we showed that no viral-derived piRNAs were produced in fruit flies during different types of viral infection. Our study provides the first evidence, to our knowledge, that the piRNA pathway does not play a major role in antiviral defense in adult Drosophila and demonstrates that viral-derived piRNA production depends on the biology of the host–virus combination rather than being part of a general antiviral process in insects.

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“…The results of the C6/36 multiplication kinetics experiments should be interpreted with caution, as this cell line was established from Aedes albopictus larva homogenates (40), which may not accurately represent the cell types and expression profiles of the mosquito midgut epithelium and other cell types associated with infection, dissemination, and transmission of JEV. The results of a recent study indicate that C6/36 cells lack a functional RNA interference (RNAi) response, suggesting that these cells may not accurately model mosquito-arbovirus interactions at the molecular level (41). However, others have demonstrated that reduced Sindbis virus (an alphavirus) replication efficiency in C6/36 cells correlated with poor experimental infection of Aedes aegypti and sharply reduced midgut infectivity and dissemination (42).…”

Section: Gi-b Emerged Inmentioning

confidence: 99%

Dynamics of the Emergence and Establishment of a Newly Dominant Genotype of Japanese Encephalitis Virus throughout Asia

Schuh¹,

Ward

Brown

et al. 2014

J Virol

124

140

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In recent years, genotype I (GI) of Japanese encephalitis virus (JEV) has displaced genotype III (GIII) as the dominant virus genotype throughout Asia. In this study, the largest collection of GIII and GI envelope gene-derived viral sequences assembled to date was used to reconstruct the spatiotemporal chronology of genotype displacement throughout Asia and to determine the evolutionary and epidemiological dynamics underlying this significant event. GI consists of two clades, GI-a and GI-b, with the latter being associated with displacement of GIII as the dominant JEV genotype throughout Asia in the 1990s. Phylogeographic analysis indicated that GI-a diverged in Thailand or Cambodia and has remained confined to tropical Asia, whereas GI-b diverged in Vietnam and then dispersed northwards to China, where it was subsequently dispersed to Japan, Korea, and Taiwan. Molecular adaptation was detected by more than one method at one site (residue 15), and coevolution was detected at two pairs of sites (residues 89 to 360 and 129 to 141) within the GI E gene protein alignment. Viral multiplication and temperature sensitivity analyses in avian and mosquito cells revealed that the GI-b isolate JE-91 had significantly higher infectivity titers in mosquito cells from 24 to 48 h postinfection than did the GI-a and GIII isolates. If the JE-91 isolate is indeed representative of GI-b, an increased multiplicative ability of GI-b viruses compared to that of GIII viruses early in mosquito infection may have resulted in a shortened extrinsic incubation period that led to an increased number of GI enzootic transmission cycles and the subsequent displacement of GIII. IMPORTANCE Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, represents the most significant etiology of childhood viral neurological infection in Asia.Despite the existence of effective vaccines, JEV is responsible for an estimated 68,000 human cases and a reported 10,000 to 15,000 deaths annually. Phylogenetic studies divided JEV into five geographically and epidemiologically distinct genotypes (GI to GV). GIII has been the source of numerous JEV epidemics throughout history and was the most frequently isolated genotype throughout most of Asia from 1935 until the 1990s. In recent years, GI has displaced GIII as the most frequently isolated virus genotype. To date, the mechanism of this genotype replacement has remained unknown. In this study, we have identified genetic determinants underlying the genotype displacement as it unfolded across Asia. JEV provides a paradigm for other flaviviruses, including West Nile, yellow fever, and dengue viruses, and the critical role of the selective advantages in the mosquito vector.

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

Cited by 286 publications

References 49 publications

Insect-Specific Viruses Detected in Laboratory Mosquito Colonies and Their Potential Implications for Experiments Evaluating Arbovirus Vector Competence

Insect-Specific Viruses Detected in Laboratory Mosquito Colonies and Their Potential Implications for Experiments Evaluating Arbovirus Vector Competence

piRNA pathway is not required for antiviral defense in Drosophila melanogaster

Dynamics of the Emergence and Establishment of a Newly Dominant Genotype of Japanese Encephalitis Virus throughout Asia

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