Isolation and characterization of exosomes released from mosquito cells infected with dengue virus

Reyes-Ruiz, José Manuel; Osuna-Ramos, Juan Fidel; Jesús-González, Luis Adrián De; Hurtado-Monzón, Arianna Mahely; Farfán-Morales, Carlos Noe; Cervantes-Salazar, Margot; Bolaños, Jeni; Cigarroa-Mayorga, O.E.; Martín‐Martínez, Eduardo San; Medina, Fernando; Fragoso‐Soriano, Rogelio; Chávez‐Munguía, Bibiana; Salas-Benito, Juan Santiago; Ángel, Rosa M. del

doi:10.1016/j.virusres.2019.03.015

Cited by 50 publications

(68 citation statements)

References 102 publications

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“…The lEVs samples were also characterized by transmission electron microscopy (TEM): The analysis of images from the mock C6/36 lEVs and ZIKV-infected C6/36 lEVs ( Figure 2D) showed different EV populations, which were heterogeneous in shape, with sizes up to 200 nm (1000 nm scale) and a defined membrane in a proper structural resolution. These data agree with other reports, which demonstrated that cells secrete EVs as a heterogeneous population with different sizes and shapes [30,44]. We did not identify viral particles inside lEVs TEMs from ZIKV-infected cells, so we evaluated the ZIKV E protein presence on the membrane's surface of lEVs.…”

Section: Zikv-infected C6/36 Cells Release Large Ev Phosphatidylserinsupporting

confidence: 90%

“…Tetraspanin-like proteins in arthropod cells, including mosquito C6/36 cells, have been described previously [16,30,[36][37][38]. Briefly, the C6/36 cells (mock and ZIKV-infected cells) were collected from the cell culture plates in sterile 1.5 mL microcentrifuge tubes, centrifuged at 550× g for 10 min at 4 • C, and separated from the medium.…”

Section: Mosquito C6/36 Cell Tetraspanin Cd63-like Protein Detection mentioning

confidence: 99%

“…After fixation, the samples were incubated with 2% osmium tetroxide (Alfa Aesar, Thermo Fisher Scientific) for 90 min at RT. The fixed pellets were washed three times with PBS and dehydrated in an ascending graded series of ethanol (30,50,70,80, 90, and 96%), including three passes (5 min each) in absolute ethanol (J.T.Baker) at RT. Three passes (5 min each) in propylene oxide (Sigma-Aldrich) at RT were then performed.…”

Section: C6/36 Evs Morphological Characterization By Transmission Elementioning

confidence: 99%

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Participation of Extracellular Vesicles from Zika-Virus-Infected Mosquito Cells in the Modification of Naïve Cells’ Behavior by Mediating Cell-to-Cell Transmission of Viral Elements

Martínez-Rojas

Quiroz-García

Monroy-Martínez

et al. 2020

Cells

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To date, no safe vaccine or antivirals for Zika virus (ZIKV) infection have been found. The pathogenesis of severe Zika, where host and viral factors participate, remains unclear. For the control of Zika, it is important to understand how ZIKV interacts with different host cells. Knowledge of the targeted cellular pathways which allow ZIKV to productively replicate and/or establish prolonged viral persistence contributes to novel vaccines and therapies. Monocytes and endothelial vascular cells are the main ZIKV targets. During the infection process, cells are capable of releasing extracellular vesicles (EVs). EVs are mediators of intercellular communication. We found that mosquito EVs released from ZIKV-infected (C6/36) cells carry viral RNA and ZIKV-E protein and are able to infect and activate naïve mosquito and mammalian cells. ZIKV C6/36 EVs promote the differentiation of naïve monocytes and induce a pro-inflammatory state with tumor necrosis factor-alpha (TNF-α) mRNA expression. ZIKV C6/36 EVs participate in endothelial vascular cell damage by inducing coagulation (TF) and inflammation (PAR-1) receptors at the endothelial surface of the cell membranes and promote a pro-inflammatory state with increased endothelial permeability. These data suggest that ZIKV C6/36 EVs may contribute to the pathogenesis of ZIKV infection in human hosts.

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Section: Zikv-infected C6/36 Cells Release Large Ev Phosphatidylserinsupporting

confidence: 90%

Section: Mosquito C6/36 Cell Tetraspanin Cd63-like Protein Detection mentioning

confidence: 99%

Section: C6/36 Evs Morphological Characterization By Transmission Elementioning

confidence: 99%

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Participation of Extracellular Vesicles from Zika-Virus-Infected Mosquito Cells in the Modification of Naïve Cells’ Behavior by Mediating Cell-to-Cell Transmission of Viral Elements

Martínez-Rojas

Quiroz-García

Monroy-Martínez

et al. 2020

Cells

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“…Progeny virus particles are considered the sole mechanism for the dissemination of flaviviruses. Nevertheless, cell-derived extracellular vesicles carrying Flavivirus vRNA have recently been shown to be infectious [111][112][113][114][115][116], thus representing an alternate mode of virus dissemination. The pathway leading to the secretion of vRNA independently of virus particles is unknown, but parallels can be drawn with the export of cellular RNAs.…”

Section: Role Of Rbps In Determining the Extracellular Fate Of Rna Momentioning

confidence: 99%

“…The intersection between cytoplasmic ribonucleoprotein complexes and endosomes might be a step prior the export of RNA molecules via exosomes propagation [142] [143]. In particular, cells infected with HCV [144][145][146], DENV [111][112][113][114], ZIKV [115] and TBEV [116] release exosomes that are infectious to recipient cells. These infectious exosomes carry full-length vRNA but do not always contain the structural viral proteins associated with virions, thus suggesting that in Flavivirus-infected cells, vRNA molecules are sorted into exosomes independently of the assembly of fully mature particles.…”

Section: Role Of Rbps In Determining the Extracellular Fate Of Rna Momentioning

confidence: 99%

Role of RNA-binding proteins during the late stages of Flavivirus replication cycle

Diosa-Toro

Prasanth

Bradrick

et al. 2020

Virol J

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The genus Flavivirus encompasses several worldwide-distributed arthropod-borne viruses including, dengue virus, Japanese encephalitis virus, West Nile virus, yellow fever virus, Zika virus, and tick-borne encephalitis virus. Infection with these viruses manifest with symptoms ranging from febrile illness to life-threatening hypotensive shock and encephalitis. Therefore, flaviviruses pose a great risk to public health. Currently, preventive measures are falling short to control epidemics and there are no antivirals against any Flavivirus. Flaviviruses carry a single stranded positive-sense RNA genome that plays multiple roles in infected cells: it is translated into viral proteins, used as template for genome replication, it is the precursor of the subgenomic flaviviral RNA and it is assembled into new virions. Furthermore, viral RNA genomes are also packaged into extracellular vesicles, e.g. exosomes, which represent an alternate mode of virus dissemination. Because RNA molecules are at the center of Flavivirus replication cycle, viral and host RNA-binding proteins (RBPs) are critical determinants of infection. Numerous studies have revealed the function of RBPs during Flavivirus infection, particularly at the level of RNA translation and replication. These proteins, however, are also critical participants at the late stages of the replication cycle. Here we revise the function of host RBPs and the viral proteins capsid, NS2A and NS3, during the packaging of viral RNA and the assembly of new virus particles. Furthermore, we go through the evidence pointing towards the importance of host RBPs in mediating cellular RNA export with the idea that the biogenesis of exosomes harboring Flavivirus RNA would follow an analogous pathway.

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Characterisation of extracellular vesicles in baculovirus infection of Spodoptera frugiperda cells

Van Es,

Possee,

King

2024

J of Extracellular Bio

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Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is an enveloped DNA virus of the Baculoviridae family. This baculovirus is widely exploited for the biological control of insect pest species and as an expression platform to produce recombinant proteins in insect cells. Extracellular vesicles (EVs) are secreted by all cells and are involved in key roles in many biological processes through their cargo consisting of proteins, RNA or DNA. In viral infections, EVs have been found to transfer both viral and cellular cargo that can elicit either a pro‐ or antiviral response in recipient cells. Here, small EVs (sEVs) released by Spodoptera frugiperda (Sf) insect cells were characterised for the first time. Using S. frugiperda (SfC1B5) cells stably expressing the baculovirus gp64, the viral envelope protein GP64 was shown to be incorporated into sEVs. Sf9 cells were also transfected with a bacmid AcMNPV genome lacking p6.9 (AcΔP6.9) to prevent budded virus production. The protein content of sEVs from both mock‐ and AcΔP6.9‐transfected cells were analysed by mass spectrometry. In addition to GP64, viral proteins Ac‐F, ME‐53 and viral ubiquitin were identified, as well as many host proteins including TSG101—which may be useful as a protein marker for sEVs.

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Isolation and characterization of exosomes released from mosquito cells infected with dengue virus

Cited by 50 publications

References 102 publications

Participation of Extracellular Vesicles from Zika-Virus-Infected Mosquito Cells in the Modification of Naïve Cells’ Behavior by Mediating Cell-to-Cell Transmission of Viral Elements

Participation of Extracellular Vesicles from Zika-Virus-Infected Mosquito Cells in the Modification of Naïve Cells’ Behavior by Mediating Cell-to-Cell Transmission of Viral Elements

Role of RNA-binding proteins during the late stages of Flavivirus replication cycle

Characterisation of extracellular vesicles in baculovirus infection of Spodoptera frugiperda cells

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