scholarly journals Mosquito Small RNA Responses to West Nile and Insect-Specific Virus Infections in Aedes and Culex Mosquito Cells

Viruses ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 271 ◽  
Author(s):  
Giel Göertz ◽  
Pascal Miesen ◽  
Gijs Overheul ◽  
Ronald van Rij ◽  
Monique van Oers ◽  
...  
Keyword(s):  
Small Rna ◽  
De Novo ◽  
Mosquito Species ◽  
Acute Infection ◽  
Mosquito Cell ◽  
Flock House Virus ◽  
Virus Infections ◽  
West Nile ◽  
Mosquito Cell Lines ◽  
Small Rna Deep Sequencing

Small RNA mediated responses are essential for antiviral defence in mosquitoes, however, they appear to differ per virus-vector combination. To further investigate the diversity of small RNA responses against viruses in mosquitoes, we applied a small RNA deep sequencing approach on five mosquito cell lines: Culex tarsalis CT cells, Aedes albopictus U4.4 and C6/36 cells, Ae. aegypti Aag2 cells (cleared from cell fusing agent virus and Culex Y virus (CYV) by repetitive dsRNA transfections) and Ae. pseudoscutellaris AP-61 cells. De novo assembly of small RNAs revealed the presence of Phasi Charoen-like virus (PCLV), Calbertado virus, Flock House virus and a novel narnavirus in CT cells, CYV in U4.4 cells, and PCLV in Aag2 cells, whereas no insect-specific viruses (ISVs) were detected in C6/36 and AP-61 cells. Next, we investigated the small RNA responses to the identified ISVs and to acute infection with the arthropod-borne West Nile virus (WNV). We demonstrate that AP-61 and C6/36 cells do not produce siRNAs to WNV infection, suggesting that AP-61, like C6/36, are Dicer-2 deficient. CT cells produced a strong siRNA response to the persistent ISVs and acute WNV infection. Interestingly, CT cells also produced viral PIWI-interacting (pi)RNAs to PCLV, but not to WNV or any of the other ISVs. In contrast, in U4.4 and Aag2 cells, WNV siRNAs, and pi-like RNAs without typical ping-pong piRNA signature were observed, while this signature was present in PCLV piRNAs in Aag2 cells. Together, our results demonstrate that mosquito small RNA responses are strongly dependent on both the mosquito cell type and/or the mosquito species and family of the infecting virus.

scholarly journals Host Adaptation of a Wolbachia Strain after Long-Term Serial Passage in Mosquito Cell Lines

2008 ◽  
Vol 74 (22) ◽  
pp. 6963-6969 ◽  
Author(s):  
Conor J. McMeniman ◽  
Amanda M. Lane ◽  
Amy W. C. Fong ◽  
Denis A. Voronin ◽  
Iñaki Iturbe-Ormaetxe ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Mosquito Species ◽  
Fruit Fly ◽  
Serial Passage ◽  
Mosquito Cell ◽  
Original Strain ◽  
New Host ◽  
Mosquito Cell Lines

ABSTRACT The horizontal transfer of the bacterium Wolbachia pipientis between invertebrate hosts hinges on the ability of Wolbachia to adapt to new intracellular environments. The experimental transfer of Wolbachia between distantly related host species often results in the loss of infection, presumably due to an inability of Wolbachia to adapt quickly to the new host. To examine the process of adaptation to a novel host, we transferred a life-shortening Wolbachia strain, wMelPop, from the fruit fly Drosophila melanogaster into a cell line derived from the mosquito Aedes albopictus. After long-term serial passage in this cell line, we transferred the mosquito-adapted wMelPop into cell lines derived from two other mosquito species, Aedes aegypti and Anopheles gambiae. After a prolonged period of serial passage in mosquito cell lines, wMelPop was reintroduced into its native host, D. melanogaster, by embryonic microinjection. The cell line-adapted wMelPop strains were characterized by a loss of infectivity when reintroduced into the original host, grew to decreased densities, and had reduced abilities to cause life-shortening infection and cytoplasmic incompatibility compared to the original strain. We interpret these shifts in phenotype as evidence for genetic adaptation to the mosquito intracellular environment. The use of cell lines to preadapt Wolbachia to novel hosts is suggested as a possible strategy to improve the success of transinfection in novel target insect species.

An integrated mosquito small RNA genomics resource reveals dynamic evolution and host responses to viruses and transposons

2020 ◽  
Author(s):  
Qicheng Ma ◽  
Satyam P. Srivastav ◽  
Stephanie Gamez ◽  
Fabiana Feitosa-Suntheimer ◽  
Edward I. Patterson ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Small Rna ◽  
Evolutionary Dynamics ◽  
Expression Profiles ◽  
Mosquito Species ◽  
Mosquito Cell ◽  
Host Responses ◽  
Dna Repeats ◽  
Small Interfering Rnas ◽  
Regulatory Pathways

ABSTRACTAlthough mosquitoes are major transmission vectors for pathogenic arboviruses, viral infection has little impact on mosquito health. This immunity is due in part to mosquito RNA interference (RNAi) pathways that generate antiviral small interfering RNAs (siRNAs) and Piwi-interacting RNAs (piRNAs). RNAi also maintains genome integrity by potently repressing mosquito transposon activity in the germline and soma. However, viral and transposon small RNA regulatory pathways have not been systematically examined together in mosquitoes. Therefore, we developed an integrated Mosquito Small RNA Genomics (MSRG) resource that analyzes the transposon and virus small RNA profiles in mosquito cell cultures and somatic and gonadal tissues across four medically important mosquito species. Our resource captures both somatic and gonadal small RNA expression profiles within mosquito cell cultures, and we report the evolutionary dynamics of a novel Mosquito-Conserved piRNA Cluster Locus (MCpiRCL) composed of satellite DNA repeats. In the larger culicine mosquito genomes we detected highly regular periodicity in piRNA biogenesis patterns coinciding with the expansion of Piwi pathway genes. Finally, our resource enables detection of crosstalk between piRNA and siRNA populations in mosquito cells during a response to virus infection. The MSRG resource will aid efforts to dissect and combat the capacity of mosquitoes to tolerate and spread arboviruses.

scholarly journals Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Raghuvir Viswanatha ◽  
Enzo Mameli ◽  
Jonathan Rodiger ◽  
Pierre Merckaert ◽  
Fabiana Feitosa-Suntheimer ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cells ◽  
Mosquito Species ◽  
Mosquito Cell ◽  
Exchange System ◽  
Public Health Burden ◽  
Cassette Exchange ◽  
Mosquito Cells ◽  
Mosquito Cell Lines ◽  
Genome Scale

AbstractMosquito-borne diseases present a worldwide public health burden. Current efforts to understand and counteract them have been aided by the use of cultured mosquito cells. Moreover, application in mammalian cells of forward genetic approaches such as CRISPR screens have identified essential genes and genes required for host-pathogen interactions, and in general, aided in functional annotation of genes. An equivalent approach for genetic screening of mosquito cell lines has been lacking. To develop such an approach, we design a new bioinformatic portal for sgRNA library design in several mosquito genomes, engineer mosquito cell lines to express Cas9 and accept sgRNA at scale, and identify optimal promoters for sgRNA expression in several mosquito species. We then optimize a recombination-mediated cassette exchange system to deliver CRISPR sgRNA and perform pooled CRISPR screens in an Anopheles cell line. Altogether, we provide a platform for high-throughput genome-scale screening in cell lines from disease vector species.

West Nile and Usutu virus infections in wild birds admitted to rehabilitation centres in Extremadura, western Spain, 2017–2019

2021 ◽  
Vol 255 ◽  
pp. 109020
Author(s):  
Daniel Bravo-Barriga ◽  
Pilar Aguilera-Sepúlveda ◽  
Fátima Guerrero-Carvajal ◽  
Francisco Llorente ◽  
David Reina ◽  
...  
Keyword(s):  
Wild Birds ◽  
Virus Infections ◽  
West Nile ◽  
Usutu Virus ◽  
Western Spain

scholarly journals Epidemiology of West Nile Virus Infections in Humans, Italy, 2012–2020: A Summary of Available Evidences

2021 ◽  
Vol 6 (2) ◽  
pp. 61
Author(s):  
Matteo Riccò ◽  
Simona Peruzzi ◽  
Federica Balzarini
Keyword(s):  
West Nile Virus ◽  
Age Groups ◽  
Invasive Disease ◽  
Incidence Rates ◽  
Annual Incidence ◽  
Virus Infections ◽  
West Nile ◽  
Po River ◽  
Air Temperatures ◽  
Sex Risk

In Italy, human cases of West Nile virus (WNV) infection have been recorded since 2008, and seasonal outbreaks have occurred almost annually. In this study, we summarize available evidences on the epidemiology of WNV and West Nile neuro-invasive disease (WNND) in humans reported between 2012 and 2020. In total, 1145 WNV infection cases were diagnosed; of them 487 (42.5%) had WNND. A significant circulation of the pathogen was suggested by studies on blood donors, with annual incidence rates ranging from 1.353 (95% confidence intervals (95% CI) 0.279–3.953) to 19.069 cases per 100,000 specimens (95% CI 13.494–26.174). The annual incidence rates of WNND increased during the study period from 0.047 cases per 100,000 (95% CI 0.031–0.068) in 2012, to 0.074 cases per 100,000 (95% CI 0.054–0.099) in 2020, peaking to 0.377 cases per 100,000 (95% CI 0.330–0.429) in 2018. There were 60 deaths. Cases of WNND were clustered in Northern Italy, particularly in the Po River Valley, during the months of August (56.7%) and September (27.5%). Higher risk for WNND was reported in subjects of male sex (risk ratio (RR) 1.545, 95% CI 1.392–1.673 compared to females), and in older age groups (RR 24.46, 95% CI 15.61–38.32 for 65–74 y.o.; RR 43.7, 95% CI 28.33–67.41 for subjects older than 75 years), while main effectors were identified in average air temperatures (incidence rate ratio (IRR) 1.3219, 95% CI 1.0053–1.7383), population density (IRR 1.0004, 95% CI 1.0001–1.0008), and occurrence of cases in the nearby provinces (IRR 1.0442, 95% CI 1.0340–1.0545). In summary, an enhanced surveillance is vital for the early detection of human cases and the prompt implementation of response measures.

scholarly journals Population dynamics of mosquito species in a West Nile virus endemic area in Madagascar

Parasite ◽  
2017 ◽  
Vol 24 ◽  
pp. 3 ◽  
Author(s):  
Luciano Michaël Tantely ◽  
Catherine Cêtre-Sossah ◽  
Tsiriniaina Rakotondranaivo ◽  
Eric Cardinale ◽  
Sébastien Boyer
Keyword(s):  
Population Dynamics ◽  
West Nile Virus ◽  
Endemic Area ◽  
Mosquito Species ◽  
West Nile

scholarly journals Environmental Predictors of Human West Nile Virus Infections, Colorado

2007 ◽  
Vol 13 (11) ◽  
pp. 1788-1790 ◽  
Author(s):  
Jennifer L. Patnaik ◽  
Lara Juliusson ◽  
Richard L. Vogt
Keyword(s):  
West Nile Virus ◽  
Virus Infections ◽  
Environmental Predictors ◽  
West Nile ◽  
Human West Nile Virus

Identification of viruses infecting sweet potato in southern China by small RNA deep sequencing and PCR detection

2018 ◽  
Vol 85 (2) ◽  
pp. 122-127 ◽  
Author(s):  
Siqi Ma ◽  
Qiufeng Zheng ◽  
Jiajie Ye ◽  
Wendi Feng ◽  
Guohui Zhou ◽  
...  
Keyword(s):  
Sweet Potato ◽  
Deep Sequencing ◽  
Small Rna ◽  
Southern China ◽  
Pcr Detection ◽  
Rna Deep Sequencing ◽  
Small Rna Deep Sequencing

scholarly journals Two Complex, Adenovirus-Based Vaccines That Together Induce Immune Responses to All Four Dengue Virus Serotypes

2006 ◽  
Vol 14 (2) ◽  
pp. 182-189 ◽  
Author(s):  
David H. Holman ◽  
Danher Wang ◽  
Kanakatte Raviprakash ◽  
Nicholas U. Raja ◽  
Min Luo ◽  
...  
Keyword(s):  
Immune Responses ◽  
Dengue Virus ◽  
Vaccine Development ◽  
De Novo ◽  
Natural Infection ◽  
Hemorrhagic Fever ◽  
Virus Infections ◽  
Dengue Vaccine ◽  
Membrane Antigens ◽  
Dengue Virus Serotypes

ABSTRACT Dengue virus infections can cause hemorrhagic fever, shock, encephalitis, and even death. Worldwide, approximately 2.5 billion people live in dengue-infested regions with about 100 million new cases each year, although many of these infections are believed to be silent. There are four antigenically distinct serotypes of dengue virus; thus, immunity from one serotype will not cross-protect from infection with the other three. The difficulties that hamper vaccine development include requirements of the natural conformation of the envelope glycoprotein to induce neutralizing immune responses and the necessity of presenting antigens of all four serotypes. Currently, the only way to meet these requirements is to use a mixture of four serotypes of live attenuated dengue viruses, but safety remains a major problem. In this study, we have developed the basis for a tetravalent dengue vaccine using a novel complex adenovirus platform that is capable of expressing multiple antigens de novo. This dengue vaccine is constructed as a pair of vectors that each expresses the premembrane and envelope genes of two different dengue virus serotypes. Upon vaccination, the vaccine expressed high levels of the dengue virus antigens in cells to mimic a natural infection and induced both humoral and cellular immune responses against multiple serotypes of dengue virus in an animal model. Further analyses show the humoral responses were indeed neutralizing against all four serotypes. Our studies demonstrate the concept of mimicking infections to induce immune responses by synthesizing dengue virus membrane antigens de novo and the feasibility of developing an effective tetravalent dengue vaccine by vector-mediated expression of glycoproteins of the four serotypes.

scholarly journals Increase in human West Nile and Usutu virus infections, Austria, 2018

2018 ◽  
Vol 23 (43) ◽  
Author(s):  
Stephan W. Aberle ◽  
Jolanta Kolodziejek ◽  
Christof Jungbauer ◽  
Karin Stiasny ◽  
Judith H. Aberle ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Blood Donors ◽  
Blood Donation ◽  
Virus Infections ◽  
West Nile Fever ◽  
West Nile ◽  
Usutu Virus ◽  
Neuroinvasive Disease ◽  
Human West Nile Virus ◽  
Imported Cases

Between 28 June and 17 September 2018, 27 cases of human West Nile virus infections were recorded in Austria; four cases of West Nile neuroinvasive disease, 11 cases of West Nile fever, six infections detected by blood donation screening and six imported cases. In addition, 18 cases of human Usutu virus infections (all blood donors) were recorded. This is the highest number of annual infections recorded in Austria since the introduction of both viruses.

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