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Background  The liver fluke  Opisthorchis viverrini  infects several million people in Southeast Asia. Adult flukes live in the bile ducts of humans, where they cause hepatobiliary pathology, including cholangiocarcinoma. Here, we investigated the potential of extracellular vesicles (EVs) secreted by the fluke and defined recombinant proteins derived from EVs to generate protective immunity in a hamster vaccination-challenge model.    Methodology/Principal findings  EVs isolated from the excretory-secretory products of  O .  viverrini  and two recombinant EV surface proteins encoding the large extracellular loops (LEL) of  Ov -TSP-2 (r Ov -TSP-2) and  Ov -TSP-3 (r Ov -TSP-3) were adjuvanted and used to vaccinate hamsters intraperitoneally followed by challenge infection with  O .  viverrini  metacercariae. The number of adult flukes recovered from hamsters immunized with EVs, r Ov -TSP-2, r Ov -TSP-3 and r Ov -TSP-2+r Ov -TSP-3 were significantly reduced compared to control animals vaccinated with adjuvant alone. The number of eggs per gram feces was also significantly reduced in hamsters vaccinated with r Ov -TSP-2 compared to controls, but no significant differences were found in the other groups. The average length of worms recovered from hamsters vaccinated with EVs, r Ov -TSP-2 and r Ov -TSP-3 was significantly shorter than that of worms recovered from the control group. Anti-EV IgG levels in serum and bile were significantly higher in hamsters vaccinated with EVs compared to control hamsters both pre- and post-challenge. In addition, levels of anti-r Ov -TSP antibodies in the serum and bile were significantly higher than control hamsters both pre- and post-challenge. Finally, antibodies against r Ov -TSP-2 and r Ov -TSP-3 blocked uptake of EVs by human primary cholangiocyte  in vitro , providing a plausible mechanism by which these vaccines exert partial efficacy and reduce the intensity of  O .  viverrini  infection.    Conclusion/Significance  Liver fluke EVs and recombinant tetraspanins derived from the EV surface when administered to hamsters induce antibody responses that block EV uptake by target bile duct cells and exert partial efficacy and against  O .  viverrini  challenge.

plos neglected tropical diseases

Vaccination of hamsters with Opisthorchis viverrini extracellular vesicles and vesicle-derived recombinant tetraspanins induces antibodies that block vesicle uptake by cholangiocytes and reduce parasite burden after challenge infection