Discrimination between exosomes and HIV-1: Purification of both vesicles from cell-free supernatants

Cantin, Réjean; Diou, Juliette; Bélanger, Danièle; Tremblay, Alexandre; Gilbert, Caroline

doi:10.1016/j.jim.2008.07.007

Cited by 280 publications

(316 citation statements)

References 97 publications

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“…At this time, it was discovered that these noninfectious viruses could be separated from their infectious counterparts (helper virus) on the basis of their slower migration in density gradients (42). Interestingly, a similar method has more recently been reported for separation of EVs from HIV virions that are produced in HEK 293 T cells or present in the plasma of HIV-1-positive individuals (44,45). Virus particles and EVs were separated on the basis of migration in velocity gradients and distinguished on the basis of the presence of p24 in virus particles and, for example, acetylcholinesterase and CD45 in EVs but not in HIV.…”

Section: Mission (mentioning

confidence: 98%

Extracellular vesicles and viruses: Are they close relatives?

Hoen

Cremer

Gallo

et al. 2016

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

425

390

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Extracellular vesicles (EVs) released by various cells are small phospholipid membrane-enclosed entities that can carry miRNA. They are now central to research in many fields of biology because they seem to constitute a new system of cell–cell communication. Physical and chemical characteristics of many EVs, as well as their biogenesis pathways, resemble those of retroviruses. Moreover, EVs generated by virus-infected cells can incorporate viral proteins and fragments of viral RNA, being thus indistinguishable from defective (noninfectious) retroviruses. EVs, depending on the proteins and genetic material incorporated in them, play a significant role in viral infection, both facilitating and suppressing it. Deciphering the mechanisms of EV-cell interactions may facilitate the design of EVs that inhibit viral infection and can be used as vehicles for targeted drug delivery.

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Section: Mission (mentioning

confidence: 98%

Extracellular vesicles and viruses: Are they close relatives?

Hoen

Cremer

Gallo

et al. 2016

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

425

390

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“…Recently, an iodixanol (Optiprep TM ) gradient was used to separate the AChE-containing exosome fraction (8.4–12%) from viruses (at 15.6%) [172]. …”

Section: Limitations and Factors That Should Be Overcome For The Thermentioning

confidence: 99%

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Yang

Hong

Cho

et al. 2018

J of Extracellular Vesicle

190

148

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Membrane proteins are of great research interest, particularly because they are rich in targets for therapeutic application. The suitability of various membrane proteins as targets for therapeutic formulations, such as drugs or antibodies, has been studied in preclinical and clinical studies. For therapeutic application, however, a protein must be expressed and purified in as close to its native conformation as possible. This has proven difficult for membrane proteins, as their native conformation requires the association with an appropriate cellular membrane. One solution to this problem is to use extracellular vesicles as a display platform. Exosomes and microvesicles are membranous extracellular vesicles that are released from most cells. Their membranes may provide a favourable microenvironment for membrane proteins to take on their proper conformation, activity, and membrane distribution; moreover, membrane proteins can cluster into microdomains on the surface of extracellular vesicles following their biogenesis. In this review, we survey the state-of-the-art of extracellular vesicle (exosome and small-sized microvesicle)-based therapeutics, evaluate the current biological understanding of these formulations, and forecast the technical advances that will be needed to continue driving the development of membrane protein therapeutics.

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“…49,50 Some studies have separated microvesicles from HIV-1 virions using (Optiprep) iodixonal gradients clearly showing the capacity to discriminate between microvesicles, virions, and cell debris. 51,52 CD45 (human leukocyte antigen) is highly expressed on the surface of vesicles derived from hematopoietic cells but is not incorporated into HIV-1 virions. 53,54 This differential incorporation of CD45 has been used to remove contaminating vesicles from virion preparations and we have used this method conversely to capture microvesicles.…”

Section: Introductionmentioning

confidence: 99%

HIV Type 1 Nef Is Released from Infected Cells in CD45⁺Microvesicles and Is Present in the Plasma of HIV-Infected Individuals

Raymond

Campbell-Sims²,

Khan³

et al. 2011

AIDS Research and Human Retroviruses

148

157

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HIV-1 Nef has been demonstrated to be integral for viral persistence, infectivity, and the acceleration of disease pathogenesis (AIDS) in humans. Nef has also been detected in the plasma of HIV-infected individuals and is released from infected cells. The form in which Nef is released from infected cells is unknown. However, Nef is a myristoylated protein and has been shown to interact with the intracellular vesicular trafficking network. Here we show that Nef is released in CD45-containing microvesicles. This microvesicular Nef (mvNef) is detected in the plasma of HIV-infected individuals at relatively high concentrations (10 ng/ml). It is also present in tissue culture supernatants of Jurkat cells infected with HIV MN . Interestingly, plasma mvNef levels in HIV þ patients did not significantly correlate with viral load or CD4 count. Microvesicular Nef levels persisted in the plasma of HIVinfected individuals despite the use of antiretroviral therapy, even in individuals with undetectable viral loads. Using cell lines, we found Nef microvesicles induce apoptosis in Jurkat T-lymphocytes but had no observed effect on the U937 monocytic cell line. Given the large amount of mvNef present in the plasma of HIV-infected individuals, the apoptotic effect of mvNef on T cells, and the observed functions of extracellular soluble Nef in vitro, it seems likely that in vivo mvNef may play a significant role in the pathogenesis of AIDS.

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Discrimination between exosomes and HIV-1: Purification of both vesicles from cell-free supernatants

Cited by 280 publications

References 97 publications

Extracellular vesicles and viruses: Are they close relatives?

Extracellular vesicles and viruses: Are they close relatives?

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

HIV Type 1 Nef Is Released from Infected Cells in CD45⁺Microvesicles and Is Present in the Plasma of HIV-Infected Individuals

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Discrimination between exosomes and HIV-1: Purification of both vesicles from cell-free supernatants

Cited by 280 publications

References 97 publications

Extracellular vesicles and viruses: Are they close relatives?

Extracellular vesicles and viruses: Are they close relatives?

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

HIV Type 1 Nef Is Released from Infected Cells in CD45+Microvesicles and Is Present in the Plasma of HIV-Infected Individuals

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HIV Type 1 Nef Is Released from Infected Cells in CD45⁺Microvesicles and Is Present in the Plasma of HIV-Infected Individuals