Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections

Montaner-Tarbes, Sergio; Novell, Elena; Tarancón, Vicens; Borràs, Francesc E.; Montoya, María; Fraile, Lorenzo; Portillo, Hernando A. del

doi:10.1038/s41598-018-36141-5

Cited by 31 publications

(29 citation statements)

References 44 publications

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“…One trial reported on extracellular vesicles from virus-infected pigs that were isolated from serum and used as an intramuscular vaccine. 11 The studies on applications of extracellular vesicles discussed in this Account are shown in Table 1.…”

Section: Surface Of Extracellular Vesiclesmentioning

confidence: 99%

Drug Delivery with Extracellular Vesicles: From Imagination to Innovation

et al. 2019

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ConspectusExtracellular vesicles are nanoparticles produced by cells. They are composed of cellular membrane with associated membrane proteins that surrounds an aqueous core containing soluble molecules such as proteins and nucleic acids, like miRNA and mRNA. They are important in many physiological and pathological processes as they can transfer biological molecules from producer cells to acceptor cells. Preparation of the niche for cancer metastasis, stimulation of tissue regeneration and orchestration of the immune response are examples of the diverse processes in which extracellular vesicles have been implicated. As a result, these vesicles have formed a source of inspiration for many scientific fields. They could be used, for example, as liquid biopsies in diagnostics, as therapeutics in regenerative medicine, or as drug delivery vehicles for transport of medicines. In this Account, we focus on drug delivery applications.As we learn more and more about these vesicles, the complexity increases. What originally appeared to be a relatively uniform population of cellular vesicles is increasingly subdivided into different subsets. Cells make various distinct vesicle types whose physicochemical aspects and composition is influenced by parental cell type, cellular activation state, local microenvironment, biogenesis pathway, and intracellular cargo sorting routes. It has proven difficult to assess the effects of changes in production protocol on the characteristics of the cell-derived vesicle population. On top of that, each isolation method for vesicles necessarily enriches certain vesicle classes and subpopulations while depleting others. Also, each method is associated with a varying degree of vesicle purity and concomitant coisolation of nonvesicular material. What emerges is a staggering heterogeneity. This constitutes one of the main challenges of the field as small changes in production and isolation protocols may have large impact on the vesicle characteristics and on subsequent vesicle activity.We try to meet this challenge by careful experimental design and development of tools that enable robust readouts. By engineering the surface and cargo of extracellular vesicles through chemical and biological techniques, favorable characteristics can be enforced while unfavorable qualities can be overruled or masked. This is coupled to the precise evaluation of the interaction of extracellular vesicles with cells to determine the extracellular vesicle uptake routes and intracellular routing. Sensitive reporter assays enable reproducible analysis of functional delivery.This systematic evaluation and optimization of extracellular vesicles improves our insight into the critical determinants of extracellular vesicle activity and should improve translation into clinical application of engineered extracellular vesicles as a new class of drug delivery systems.

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Section: Surface Of Extracellular Vesiclesmentioning

confidence: 99%

Drug Delivery with Extracellular Vesicles: From Imagination to Innovation

et al. 2019

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“…Recent research works have made contributions here by exploring the proteomics of the viral particles and intracellular proteome of this virus [3,63]. Our work was focused on extracellular vesicle-enriched fractions, as these have been shown to be important for several viral infections, and recently they have been used as a vaccination strategy for another important swine disease—PRRS virus [22].…”

Section: Resultsmentioning

confidence: 99%

“…On the other hand, extracellular vesicles (EVs) have proven to be a promising new vaccination platform for veterinary diseases in situations in which conventional approaches have not been completely successful, such as PRRSV [21,22,23,24]. EVs are small, round vesicles of 50–400 nm in diameter which are secreted by different cell types, and they are classified as microvesicles if formed by the direct budding of the plasma membrane or exosomes if derived from late endosome trafficking and multivesicular body formation before final release to the extracellular space [25,26].…”

Section: Introductionmentioning

confidence: 99%

Serum-Derived Extracellular Vesicles from African Swine Fever Virus-Infected Pigs Selectively Recruit Viral and Porcine Proteins

Montaner-Tarbes¹,

Pujol

Jabbar

et al. 2019

Viruses

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African swine fever is a devastating hemorrhagic infectious disease, which affects domestic and wild swines (Sus scrofa) of all breeds and ages, with a high lethality of up to 90–100% in naïve animals. The causative agent, African swine fever virus (ASFV), is a large and complex double-stranded DNA arbovirus which is currently spreading worldwide, with serious socioeconomic consequences. There is no treatment or effective vaccine commercially available, and most of the current research is focused on attenuated viral models, with limited success so far. Thus, new strategies are under investigation. Extracellular vesicles (EVs) have proven to be a promising new vaccination platform for veterinary diseases in situations in which conventional approaches have not been completely successful. Here, serum extracellular vesicles from infected pigs using two different ASFV viruses (OURT 88/3 and Benin ΔMGF), corresponding to a naturally attenuated virus and a deletion mutant, respectively, were characterized in order to determine possible differences in the content of swine and viral proteins in EV-enriched fractions. Firstly, EVs were characterized by their CD5, CD63, CD81 and CD163 surface expression. Secondly, ASFV proteins were detected on the surface of EVs from ASFV-infected pig serum. Finally, proteomic analysis revealed few specific proteins from ASFV in the EVs, but 942 swine proteins were detected in all EV preparations (negative controls, and OURT 88/3 and Benin ΔMGF-infected preparations). However, in samples from OURT 88/3-infected animals, only a small number of proteins were differentially identified compared to control uninfected animals. Fifty-six swine proteins (Group Benin) and seven proteins (Group OURT 88/3) were differentially detected on EVs when compared to the EV control group. Most of these were related to coagulation cascades. The results presented here could contribute to a better understanding of ASFV pathogenesis and immune/protective responses in the host.

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“…Frequencies of IFN‐γ secreting cells were measured as described before [19]. The 96‐well PVDF‐based membrane plates were coated with 15 µg/mL monoclonal antibody overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Green preparation of hydrogel particles‐in‐emulsions for simultaneous enhancement of humoral and cell‐mediated immunity

Zou

Ngai

et al. 2020

Engineering in Life Sciences

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Emulsions are one of the most often used vaccine adjuvant formulations. Although they promote high humoral immunity, the induced cellular immunity is often poor, which restrict their application. To enhance the cellular immunity, some researchers have prepared mixed formulations by adding particles into the aqueous phase of emulsions. However, the particle preparation process usually involves the addition and removal of organic reagents, which is environmentally unfriendly and cumbersome. Moreover, the obtained vaccine adjuvant only induces limited cell‐mediated immunity and humoral immunity compared with emulsion‐adjuvanted vaccines. Herein, we developed a green and simple method for fabricating a novel nanoparticles‐in‐emulsions (NPE) formulation. Firstly, a temperature‐sensitive hydrogel was used to prepare particles by self‐solidification without additional crosslinking reagents. Secondly, the white oil was used as organic phase to avoid the particle washing procedures and organic solvent residues. Moreover, the effect of NPE as vaccine adjuvant was evaluated by using two veterinary vaccines as model antigens. NPE showed advantages than the conventional vaccine formulations in inducing both humoral and cellular immunity. This work provides a facile and broadly applicable approach for preparing nanoparticles‐in‐emulsions formulation, and presents an effective adjuvant for enhancing immunity against infectious diseases.

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Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections

Cited by 31 publications

References 44 publications

Drug Delivery with Extracellular Vesicles: From Imagination to Innovation

Drug Delivery with Extracellular Vesicles: From Imagination to Innovation

Serum-Derived Extracellular Vesicles from African Swine Fever Virus-Infected Pigs Selectively Recruit Viral and Porcine Proteins

Green preparation of hydrogel particles‐in‐emulsions for simultaneous enhancement of humoral and cell‐mediated immunity

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