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.