Comparison of exosomes purified via ultracentrifugation (UC) and Total Exosome Isolation (TEI) reagent from the serum of Marek’s disease virus (MDV)-vaccinated and tumor-bearing chickens

Neerukonda, Sabari Nath; Egan, Nicholas; Patria, Joseph; Assakhi, Imane; Tavlarides-Hontz, Phaedra; Muñoz, Eric R.; Hudson, Matthew B.; Parcells, Mark S.

doi:10.1016/j.jviromet.2018.10.004

Cited by 28 publications

(26 citation statements)

References 46 publications

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“…Our analyses indicated the presence of variable numbers of round and irregularly-shaped vesicles of approximately 50–70 nm in size that displayed exosome morphology (Figure 1D). These vesicles were occasionally observed as dense aggregates and were consistent with our previous observations [22].…”

Section: Resultssupporting

confidence: 92%

“…Exosome purification was carried out using total exosome isolation reagent (TEI, Invitrogen, Carlsbad, CA, USA) solution according to the manufacturer’s recommendations as described previously [22]. For small RNA sequencing experiments, serum exosomes were purified by ExoQuick Solution (System Biosciences, Palo Alto, CA, USA), following manufacturer’s recommendations.…”

Section: Methodsmentioning

confidence: 99%

“…The concentration and size distribution profiles of exosome particles were evaluated by tracking their Brownian motion using a Nanosight nanoparticle tracking instrument (NS300, Malvern, Worcestershire, UK) and analyzed with NTA 3.2 Dev Build 3.2.16 software, essentially as described [22]. The following post-acquisition analysis settings were selected: minimum detection threshold 4, automatic blur, and automatic minimum expected particle size.…”

Section: Methodsmentioning

confidence: 99%

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Comparison of the Transcriptomes and Proteomes of Serum Exosomes from Marek’s Disease Virus-Vaccinated and Protected and Lymphoma-Bearing Chickens

Neerukonda

Tavlarides-Hontz

McCarthy

et al. 2019

Genes

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Marek’s disease virus (MDV) is the causative agent of Marek’s disease (MD), a complex pathology of chickens characterized by paralysis, immunosuppression, and T-cell lymphomagenesis. MD is controlled in poultry production via vaccines administered in ovo or at hatch, and these confer protection against lymphoma formation, but not superinfection by MDV field strains. Despite vaccine-induced humoral and cell-mediated immune responses, mechanisms eliciting systemic protection remain unclear. Here we report the contents of serum exosomes to assess their possible roles as indicators of systemic immunity, and alternatively, tumor formation. We examined the RNA and protein content of serum exosomes from CVI988 (Rispens)-vaccinated and protected chickens (VEX), and unvaccinated tumor-bearing chickens (TEX), via deep-sequencing and mass spectrometry, respectively. Bioinformatic analyses of microRNAs (miRNAs) and predicted miRNA targets indicated a greater abundance of tumor suppressor miRNAs in VEX compared to TEX. Conversely, oncomiRs originating from cellular (miRs 106a-363) and MDV miRNA clusters were more abundant in TEX compared to VEX. Most notably, mRNAs mapping to the entire MDV genome were identified in VEX, while mRNAs mapping to the repeats flanking the unique long (IRL/TRL) were identified in TEX. These data suggest that long-term systemic vaccine-induced immune responses may be mediated at the level of VEX which transfer viral mRNAs to antigen presenting cells systemically. Proteomic analyses of these exosomes suggested potential biomarkers for VEX and TEX. These data provide important putative insight into MDV-mediated immune suppression and vaccine responses, as well as potential serum biomarkers for MD protection and susceptibility.

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Section: Resultssupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Comparison of the Transcriptomes and Proteomes of Serum Exosomes from Marek’s Disease Virus-Vaccinated and Protected and Lymphoma-Bearing Chickens

Neerukonda

Tavlarides-Hontz

McCarthy

et al. 2019

Genes

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“…The mean size and size distribution profile of exosomes that were isolated and purified from SVV-infected or control-treated IBRS-2 cell culture supernatants were analyzed as previously described [15,16]. In brief, the samples were diluted at a ratio of 1:1000 in PBS containing 0.05% Tween-20 in a total volume of 1.0 mL.…”

Section: Nanoparticle Tracking Analysis (Nta)mentioning

confidence: 99%

Intercellular transmission of Seneca Valley virus mediated by exosomes

Shi

et al. 2020

Vet Res

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Seneca Valley virus (SVV) is a non-encapsulated single-stranded positive-strand RNA virus whose transmission routes have not yet been fully elucidated. Exosomes have been implicated in the intercellular transport of a variety of materials, such as proteins, RNA, and liposomes. However, whether exosomes can mediate SVV intercellular transmission remains unknown. In this study, we extracted exosomes from SVV-infected IBRS-2 cells to investigate intercellular transmission. Our results suggest that the intercellular transmission of SVV is mediated by exosomes. The results of co-localization and RT-qPCR studies showed that exosomes harbor SVV and enable the virus to proliferate in both susceptible and non-susceptible cells. Furthermore, the replication of SVV was inhibited when IBRS-2 cells were treated with interfering RNA Rab27a and exosome inhibitor GW4869. Finally, neutralization experiments were performed to further verify whether the virus was encapsulated by the exosomes that mediated transmission between cells. It was found that exosome-mediated intercellular transmission was not blocked by SVV-specific neutralizing antibodies. This study reveals a new transmission route of SVV and provides clear evidence regarding the pathogenesis of SVV, information which can also be useful for identifying therapeutic interventions.

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“…When detecting the expression of various miRNAs within exosomes, researchers observed that exosomes from vaccinated chickens exhibited greater expression of tumor suppressor miRNA (gga-miR-146b) and less expression of oncomiR (gga-miR-21) compared to those obtained from unvaccinated controls. Thus, gga-miR-146b and -21 may serve as serum exosome biomarkers for vaccine-induced protection and for MD tumors, respectively (Nath Neerukonda et al, 2019).…”

Section: Mirna and Marek's Diseasementioning

confidence: 99%

Research Progress on MicroRNAs Involved in the Regulation of Chicken Diseases

Wang

2020

J. Poult. Sci.

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MicroRNAs (miRNAs) are small, non-coding RNA molecules that inhibit protein translation from target mRNAs. Accumulating evidence suggests that miRNAs can regulate a broad range of biological pathways, including cell differentiation, apoptosis, and carcinogenesis. With the development of miRNAs, the investigation of miRNA functions has emerged as a hot research field. Due to the intensive farming in recent decades, chickens are easily influenced by various pathogen transmissions, and this has resulted in large economic losses. Recent reports have shown that miRNAs can play critical roles in the regulation of chicken diseases. Therefore, the aim of this review is to briefly discuss the current knowledge regarding the effects of miRNAs on chickens suffering from common viral diseases, mycoplasmosis, necrotic enteritis, and ovarian tumors. Additionally, the detailed targets of miRNAs and their possible functions are also summarized. This review intends to highlight the key role of miRNAs in regard to chickens and presents the possibility of improving chicken disease resistance through the regulation of miRNAs.

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Comparison of exosomes purified via ultracentrifugation (UC) and Total Exosome Isolation (TEI) reagent from the serum of Marek’s disease virus (MDV)-vaccinated and tumor-bearing chickens

Cited by 28 publications

References 46 publications

Comparison of the Transcriptomes and Proteomes of Serum Exosomes from Marek’s Disease Virus-Vaccinated and Protected and Lymphoma-Bearing Chickens

Comparison of the Transcriptomes and Proteomes of Serum Exosomes from Marek’s Disease Virus-Vaccinated and Protected and Lymphoma-Bearing Chickens

Intercellular transmission of Seneca Valley virus mediated by exosomes

Research Progress on MicroRNAs Involved in the Regulation of Chicken Diseases

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