The anti-immune dengue subgenomic flaviviral RNA is found in vesicles in mosquito saliva and associated with increased infectivity

2021 ◽  
Author(s):  
Shih-Chia Yeh ◽  
Wei-Lian Tan ◽  
Avisha Chowdhury ◽  
Vanessa Chuo ◽  
R. Manjunatha Kini ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Dengue Virus ◽  
Extracellular Vesicles ◽  
Virus Transmission ◽  
Human Cells ◽  
Infected Mosquito ◽  
Virus Infectivity ◽  
Dengue Viruses ◽  
Mosquito Saliva

AbstractMosquito transmission of dengue viruses to humans starts with infection of skin resident cells at the biting site. There is great interest in identifying transmission-enhancing factors in mosquito saliva in order to counteract them. Here we report the discovery of high levels of subgenomic flaviviral RNA (sfRNA) in dengue virus 2-infected mosquito saliva. We show that salivary sfRNA is protected in detergent-sensitive, protease-resistant compartments. Furthermore, we show that incubation with mosquito saliva containing higher sfRNA levels results in higher virus infectivity in human cells. Since sfRNA potently inhibits innate immunity in human cells, we posit that sfRNA in mosquito saliva is present in extracellular vesicles that deliver it to cells at the biting site to inhibit innate immunity and enhance dengue virus transmission.

scholarly journals Dengue Virus Infection of Aedes aegypti Alters Extracellular Vesicle Protein Cargo to Enhance Virus Transmission

2020 ◽  
Vol 21 (18) ◽  
pp. 6609
Author(s):  
Alexander S. Gold ◽  
Fabiana Feitosa-Suntheimer ◽  
Ricardo V. Araujo ◽  
Ryan M. Hekman ◽  
Sultan Asad ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Extracellular Vesicles ◽  
High Performance ◽  
Dengue Infection ◽  
Virus Transmission ◽  
Viral Disease ◽  
Extracellular Vesicle ◽  
Transmission Cycle ◽  
Packaging Strategy

Dengue is the most burdensome vector-borne viral disease in the world. Dengue virus (DENV), the etiological cause of dengue, is transmitted primarily by the Aedes aegypti mosquito. Like any arbovirus, the transmission cycle of dengue involves the complex interactions of a multitude of human and mosquito factors. One point during this transmission cycle that is rich in these interactions is the biting event by the mosquito, upon which its saliva is injected into the host. A number of components in mosquito saliva have been shown to play a pivotal role in the transmission of dengue, however one such component that is not as well characterized is extracellular vesicles. Here, using high-performance liquid chromatography in tandem with mass spectrometry, we show that dengue infection altered the protein cargo of Aedes aegypti extracellular vesicles, resulting in the packaging of proteins with infection-enhancing ability. Our results support the presence of an infection-dependent pro-viral protein packaging strategy that uses the differential packaging of pro-viral proteins in extracellular vesicles of Ae. aegypti saliva to promote transmission. These studies represent the first investigation into the function of Ae. aegypti extracellular vesicle cargo during dengue infection.

scholarly journals Arthropod EVs mediate dengue virus transmission through interaction with a tetraspanin domain containing glycoprotein Tsp29Fb

2018 ◽  
Vol 115 (28) ◽  
pp. E6604-E6613 ◽  
Author(s):  
Ashish Vora ◽  
Wenshuo Zhou ◽  
Berlin Londono-Renteria ◽  
Michael Woodson ◽  
Michael B. Sherman ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Mammalian Cells ◽  
Virus Transmission ◽  
Direct Interaction ◽  
Rnase A ◽  
Human Cells ◽  
Viral Rna ◽  
E Protein ◽  
In Vitro Cell Lines

Dengue virus (DENV) is a mosquito-borne flavivirus that causes dengue fever in humans, worldwide. Using in vitro cell lines derived from Aedes albopictus and Aedes aegypti, the primary vectors of DENV, we report that DENV2/DENV3-infected cells secrete extracellular vesicles (EVs), including exosomes, containing infectious viral RNA and proteins. A full-length DENV2 genome, detected in arthropod EVs, was infectious to naïve mosquito and mammalian cells, including human-skin keratinocytes and blood endothelial cells. Cryo-electron microscopy showed mosquito EVs with a size range from 30 to 250 nm. Treatments with RNase A, Triton X-100, and 4G2 antibody-bead binding assays showed that infectious DENV2-RNA and proteins are contained inside EVs. Viral plaque formation and dilution assays also showed securely contained infectious viral RNA and proteins in EVs are transmitted to human cells. Up-regulated HSP70 upon DENV2 infection showed no role in viral replication and transmission through EVs. In addition, qRT-PCR and immunoblotting results revealed that DENV2 up-regulates expression of a mosquito tetraspanin-domain–containing glycoprotein, designated as Tsp29Fb, in A. aegypti mosquitoes, cells, and EVs. RNAi-mediated silencing and antibody blocking of Tsp29Fb resulted in reduced DENV2 loads in both mosquito cells and EVs. Immunoprecipitation showed Tsp29Fb to directly interact with DENV2 E-protein. Furthermore, treatment with GW4869 (exosome-release inhibitor) affected viral burden, direct interaction of Tsp29Fb with E-protein and EV-mediated transmission of viral RNA and proteins to naïve human cells. In summary, we report a very important finding on EV-mediated transmission of DENV2 from arthropod to mammalian cells through interactions with an arthropod EVs-enriched marker Tsp29Fb.

The effects of mosquito saliva on dengue virus infectivity in humans

2016 ◽  
Vol 21 ◽  
pp. 139-145 ◽  
Author(s):  
Sineewanlaya Wichit ◽  
Pauline Ferraris ◽  
Valérie Choumet ◽  
Dorothée Missé
Keyword(s):  
Dengue Virus ◽  
Virus Infectivity ◽  
Mosquito Saliva

scholarly journals A phosphomimetic-based mechanism of dengue virus to antagonize innate immunity

2016 ◽  
Vol 17 (5) ◽  
pp. 523-530 ◽  
Author(s):  
Ying Kai Chan ◽  
Michaela U Gack
Keyword(s):  
Innate Immunity ◽  
Dengue Virus

scholarly journals Dengue Virus Transmission from Donor to Recipient during Haploidentical Stem Cell Transplantation

IDCases ◽  
2021 ◽  
pp. e01220
Author(s):  
Anjali Yadav ◽  
Neha Rastogi ◽  
K. Upasana ◽  
Sunisha Arora ◽  
Dhwanee Thakkar ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Dengue Virus ◽  
Cell Transplantation ◽  
Virus Transmission ◽  
Haploidentical Stem Cell Transplantation

P122 Dengue virus targets the adaptor protein MITA to subvert host innate immunity

Cytokine ◽  
2012 ◽  
Vol 59 (3) ◽  
pp. 558 ◽  
Author(s):  
C.-Y. Yu ◽  
T.-H. Chang ◽  
J.-J. Liang ◽  
R.-L. Chiang ◽  
Y.-L. Lee ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Dengue Virus ◽  
Adaptor Protein ◽  
Host Innate Immunity

scholarly journals Delivery of functional exogenous proteins by plant-derived vesicles to human cells in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Luiza Garaeva ◽  
Roman Kamyshinsky ◽  
Yury Kil ◽  
Elena Varfolomeeva ◽  
Nikolai Verlov ◽  
...  
Keyword(s):  
Cell Culture ◽  
Colon Cancer ◽  
Extracellular Vesicles ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Human Cells ◽  
Bioactive Molecules ◽  
Native Plant ◽  
Peripheral Blood Mononuclear

AbstractPlant-derived extracellular vesicles (EVs) gain more and more attention as promising carriers of exogenous bioactive molecules to the human cells. Derived from various edible sources, these EVs are remarkably biocompatible, biodegradable and highly abundant from plants. In this work, EVs from grapefruit juice were isolated by differential centrifugation followed by characterization of their size, quantity and morphology by nanoparticle tracking analysis, dynamic light scattering, atomic force microscopy and cryo-electron microscopy (Cryo-EM). In Cryo-EM experiments, we visualized grapefruit EVs with the average size of 41 ± 13 nm, confirmed their round-shaped morphology and estimated the thickness of their lipid bilayer as 5.3 ± 0.8 nm. Further, using cell culture models, we have successfully demonstrated that native grapefruit-derived extracellular vesicles (GF-EVs) are highly efficient carriers for the delivery of the exogenous Alexa Fluor 647 labeled bovine serum albumin (BSA) and heat shock protein 70 (HSP70) into both human peripheral blood mononuclear cells and colon cancer cells. Interestingly, loading to plant EVs significantly ameliorated the uptake of exogenous proteins by human cells compared to the same proteins without EVs. Most importantly, we have confirmed the functional activity of human recombinant HSP70 in the colon cancer cell culture upon delivery by GF-EVs. Analysis of the biodistribution of GF-EVs loaded with 125I-labeled BSA in mice demonstrated a significant uptake of the grapefruit-derived extracellular vesicles by the majority of organs. The results of our study indicate that native plant EVs might be safe and effective carriers of exogenous proteins into human cells.

scholarly journals Natural vertical transmission by Stegomyia albopicta as dengue vector in Brazil

2009 ◽  
Vol 69 (1) ◽  
pp. 123-127 ◽  
Author(s):  
AB. Cecílio ◽  
ES. Campanelli ◽  
KPR. Souza ◽  
LB. Figueiredo ◽  
MC. Resende
Keyword(s):  
Dengue Virus ◽  
Vertical Transmission ◽  
Dengue Vector ◽  
Infection Rates ◽  
Dengue Viruses ◽  
Stegomyia Albopicta ◽  
First Time ◽  
Minimum Infection Rate ◽  
Dengue Virus Type 2

The mosquito Stegomyia albopicta is among the most important arbovirus vectors in the world, particularly for Dengue viruses. Their natural history suggests that biologically these viruses are highly adapted to their mosquito hosts and they were most likely mosquito viruses prior to becoming adapted to lower primates and humans. As well as being maintained by transmission among susceptible humans, Dengue viruses may also be maintained by vertical transmission in mosquitoes during inter-epidemic periods. The larvae and mosquitoes of Stegomyia albopicta were used to identify the vertical transmission of the dengue virus in nature and to confirm the vectorial capacity concerning the Dengue virus type 2 infection. The minimum infection rate concerning S. albopicta infection with the Dengue virus was 1:36.45. In Brazil this was the first time that high minimum infection rates of vertical transmission of S. albopicta were detected in this species.

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