2016
DOI: 10.1038/srep29425
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Biochemical and proteomic characterization of retrovirus Gag based microparticles carrying melanoma antigens

Abstract: Extracellular vesicles are membraneous particles released by a variety of cells into the extracellular microenvironment. Retroviruses utilize the cellular vesiculation pathway for virus budding/assembly and the retrovirus Gag protein induces the spontaneous formation of microvesicles or virus-like particles (VLPs) when expressed in the mammalian cells. In this study, five different melanoma antigens, MAGEA4, MAGEA10, MART1, TRP1 and MCAM, were incorporated into the VLPs and their localization within the partic… Show more

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Cited by 14 publications
(37 citation statements)
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“…We recently showed that MAGE4 and MAGEA10 proteins are efficiently incorporated into retrovirus Gag protein induced virus-like particles (VLPs) [28]. In the current study, we show that MAGEA4 and MAGEA10 proteins are incorporated into naturally occurring EVs released by mouse fibroblast and human osteosarcoma U2OS cells and are expressed on the surface of EVs.…”
Section: Introductionmentioning
confidence: 54%
See 1 more Smart Citation
“…We recently showed that MAGE4 and MAGEA10 proteins are efficiently incorporated into retrovirus Gag protein induced virus-like particles (VLPs) [28]. In the current study, we show that MAGEA4 and MAGEA10 proteins are incorporated into naturally occurring EVs released by mouse fibroblast and human osteosarcoma U2OS cells and are expressed on the surface of EVs.…”
Section: Introductionmentioning
confidence: 54%
“…In order to determine whether MAGEA proteins are expressed on the surface of EVs, we performed the immunostaining analysis of EVs bound to aldehyde/sulfate latex beads [28]. As shown in Figure 2A, MAGEA4 and MAGEA10 specific antibodies recognized all three subtypes of EVs by flow cytometer analysis suggesting that at least part of these proteins are exposed on the surface of EVs.…”
Section: Resultsmentioning
confidence: 99%
“…Protein pellets were processed into peptides as described previously 83 . Samples were injected to an Agilent 1200 series nano-LC with an in-house packed (3 µm ReproSil-Pur C18AQ particles) 15 cm × 75 µm ID emitter-column (New Objective).…”
Section: Methodsmentioning
confidence: 99%
“…MAGEA4 and MAGEA10 coding sequences from pQMCF-MAGEA4 and pQMCF-MAGEA10 vectors ( 19 ) were cloned in frame into pET28a vector using NheI restriction enzyme to fuse the coding sequence with His-tag. Recombinant His-tagged MAGEA4 and MAGEA10 proteins were transformed into Escherichia coli ( E coli ) cells BL-CodonPlus ™ RP (Invitrogen; USA); transformed bacteria were grown at 37°C to the spectrophotometric density 0.6 (OD 600 nm; Ultraspec 7000; GE Healthcare Life Sciences, Little Chalfont, UK) and induced with 1 mM IPTG for 2 h at room temperature.…”
Section: Methodsmentioning
confidence: 99%
“…MAGEA subfamily proteins are highly conserved and it is very difficult to get antibodies that recognize only one member of the family specifically. For example, MAGEA4 and MAGEA10 proteins share more than 50% sequence identity on the amino acid level, but have different sizes and cellular localizations ( 19 ). Several antibodies used in immunohistochemical studies cross-react with many MAGEA proteins and have been seen in multiple cancer types to localize both in the cytoplasm and in the nucleus ( 20 22 ).…”
Section: Introductionmentioning
confidence: 99%