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

Figures

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  Figure 1. Rectal temperatures in pigs. Rectal temperatures were assessed at different days after immunization with two different African swine fever virus (ASFV) strains, (a) OURT-88/3 and (b) deletion mutant BeninDMFG, by an intramuscular route. Before inoculation, rectal temperatures of pigs ranged between 39 and 39.9 °C. At day 24 post immunization, the rectal temperatures of pigs immunized with OURT88/3 ranged between 39.4 and 39.9 °C, while in the group of pigs immunized with Benin ΔMGF, temperatures ranged between 38.7 and 39.6 °C.

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  Figure 2. Characterization of extracellular vesicles derived from uninfected and ASFV infected swine sera. (a,d) FACS analysis of extracellular vesicle (EV) fractions derived from infected and control animals; (b,e) size distribution as measured by electron microscopy; (c,f) transmission electron microscopy of negatively-stained uninfected and infected samples, respectively. EVs are shown with red arrows (scale bar in c: 200 nm; scale bar in f: 500 nm).

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  Figure 3. FACS analysis of EV-enriched fractions derived from infected and control animals. (a) EVs from representative sera of an infected animal with OURT 88/3 at 24 days post infection, (b) EV profiles from a representative of uninfected swine sera, (c) EV FACS profile from a representative animal infected with Benin ΔMGF virus. In all samples, molecular markers CD5 and CD163 were used as control markers for EVs and three different viral proteins were evaluated (p30, p54 and p72).

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  Figure 4. Swine proteins on EV-enriched fractions evaluated at 24 days post infection (red proteins were significant at p < 0.05) and above the volcano plot threshold (p < 0.01). (a) Proteins differentially expressed in EV-enriched fractions from Group A (infected with OURT 88/3). (b) Proteins differentially expressed in EV-enriched fractions from Group B (infected with Benin ΔMGF).

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  Figure 5. Venn diagram for the comparison of significantly expressed swine proteins in a virus-dependent manner detected in serum-derived EVs.

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  Figure 6. Gene ontology analyses of Benin ΔMGF EV-enriched fractions, where differentially expressed swine proteins were classified according to the biological process, cellular component or molecular function, as well as a string pathway analysis for all proteins differentially expressed in Benin ΔMGF-infected animals.