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

Overview

Journal PLoS Negl Trop Dis

Specialties Infectious Diseases
Tropical Medicine

Date 2019 May 29

PMID 31136572

Citations 28

Authors

Sujittra Chaiyadet

Javier Sotillo

Watchara Krueajampa

Sophita Thongsen

Paul J Brindley

Banchob Sripa

Alex Loukas

Thewarach Laha

Affiliations

Soon will be listed here.

Abstract

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 (rOv-TSP-2) and Ov-TSP-3 (rOv-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, rOv-TSP-2, rOv-TSP-3 and rOv-TSP-2+rOv-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 rOv-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, rOv-TSP-2 and rOv-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-rOv-TSP antibodies in the serum and bile were significantly higher than control hamsters both pre- and post-challenge. Finally, antibodies against rOv-TSP-2 and rOv-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.

Citing Articles

Secreted and surface proteome and transcriptome of .

Wong Y, Pearson M, Fedorova O, Ivanov V, Khmelevskaya E, Tedla B Front Parasitol. 2025; 2():1195457.

PMID: 39816815 PMC: 11732047. DOI: 10.3389/fpara.2023.1195457.

Helminth-derived molecules: pathogenic and pharmacopeial roles.

Zhang Y, Shen C, Zhu X, Leow C, Ji M, Xu Z J Biomed Res. 2024; :1-22.

PMID: 39314046 PMC: 11629161. DOI: 10.7555/JBR.38.20240177.

Development of a subunit vaccine against the cholangiocarcinoma causing : a computational approach.

Shah M, Sitara F, Sarfraz A, Shehroz M, Wara T, Perveen A Front Immunol. 2024; 15:1281544.

PMID: 39050853 PMC: 11266093. DOI: 10.3389/fimmu.2024.1281544.

Guidelines for the purification and characterization of extracellular vesicles of parasites.

Fernandez-Becerra C, Xander P, Alfandari D, Dong G, Aparici-Herraiz I, Rosenhek-Goldian I J Extracell Biol. 2024; 2(10):e117.

PMID: 38939734 PMC: 11080789. DOI: 10.1002/jex2.117.

Opisthorchis viverrini excretory-secretory products suppress GLUT8 of cholangiocytes.

Kha S, Chaiyadet S, Saichua P, Tangkawatana S, Sripa B, Suttiprapa S Parasitol Res. 2024; 123(3):161.

PMID: 38491300 DOI: 10.1007/s00436-024-08184-3.

References

Buck A, Coakley G, Simbari F, McSorley H, Quintana J, Le Bihan T . Exosomes secreted by nematode parasites transfer small RNAs to mammalian cells and modulate innate immunity. Nat Commun. 2014; 5:5488. PMC: 4263141. DOI: 10.1038/ncomms6488. View

Chaiyadet S, Smout M, Johnson M, Whitchurch C, Turnbull L, Kaewkes S . Excretory/secretory products of the carcinogenic liver fluke are endocytosed by human cholangiocytes and drive cell proliferation and IL6 production. Int J Parasitol. 2015; 45(12):773-81. PMC: 4912216. DOI: 10.1016/j.ijpara.2015.06.001. View

Merrifield M, Hotez P, Beaumier C, Gillespie P, Strych U, Hayward T . Advancing a vaccine to prevent human schistosomiasis. Vaccine. 2016; 34(26):2988-2991. DOI: 10.1016/j.vaccine.2016.03.079. View

Sripa B, Brindley P, Mulvenna J, Laha T, Smout M, Mairiang E . The tumorigenic liver fluke Opisthorchis viverrini--multiple pathways to cancer. Trends Parasitol. 2012; 28(10):395-407. PMC: 3682777. DOI: 10.1016/j.pt.2012.07.006. View

Nowacki F, Swain M, Klychnikov O, Niazi U, Ivens A, Quintana J . Protein and small non-coding RNA-enriched extracellular vesicles are released by the pathogenic blood fluke Schistosoma mansoni. J Extracell Vesicles. 2015; 4:28665. PMC: 4595467. DOI: 10.3402/jev.v4.28665. View

Chaiyadet S, Sotillo J, Smout M, Cantacessi C, Jones M, Johnson M . Carcinogenic Liver Fluke Secretes Extracellular Vesicles That Promote Cholangiocytes to Adopt a Tumorigenic Phenotype. J Infect Dis. 2015; 212(10):1636-45. PMC: 4621255. DOI: 10.1093/infdis/jiv291. View

Sithithaworn P, Andrews R, Nguyen V, Wongsaroj T, Sinuon M, Odermatt P . The current status of opisthorchiasis and clonorchiasis in the Mekong Basin. Parasitol Int. 2011; 61(1):10-6. PMC: 3836690. DOI: 10.1016/j.parint.2011.08.014. View

Raposo G, Stoorvogel W . Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol. 2013; 200(4):373-83. PMC: 3575529. DOI: 10.1083/jcb.201211138. View

Jittimanee J, Sermswan R, Kaewraemruaen C, Barta J, MacInnes J, Maleewong W . Protective immunization of hamsters against Opisthorchis viverrini infection is associated with the reduction of TGF-β expression. Acta Trop. 2012; 122(2):189-95. DOI: 10.1016/j.actatropica.2012.01.010. View

10.

Tran M, Pearson M, Bethony J, Smyth D, Jones M, Duke M . Tetraspanins on the surface of Schistosoma mansoni are protective antigens against schistosomiasis. Nat Med. 2006; 12(7):835-40. DOI: 10.1038/nm1430. View

11.

Zhu L, Liu J, Dao J, Lu K, Li H, Gu H . Molecular characterization of S. japonicum exosome-like vesicles reveals their regulatory roles in parasite-host interactions. Sci Rep. 2016; 6:25885. PMC: 4865838. DOI: 10.1038/srep25885. View

12.

Laha T, Pinlaor P, Mulvenna J, Sripa B, Sripa M, Smout M . Gene discovery for the carcinogenic human liver fluke, Opisthorchis viverrini. BMC Genomics. 2007; 8:189. PMC: 1913519. DOI: 10.1186/1471-2164-8-189. View

13.

Haswell-Elkins M, Sithithaworn P, Mairiang E, Elkins D, Wongratanacheewin S, Kaewkes S . Immune responsiveness and parasite-specific antibody levels in human hepatobiliary disease associated with Opisthorchis viverrini infection. Clin Exp Immunol. 1991; 84(2):213-8. PMC: 1535388. DOI: 10.1111/j.1365-2249.1991.tb08151.x. View

14.

Sriamporn S, Pisani P, Pipitgool V, Suwanrungruang K, Kamsa-Ard S, Parkin D . Prevalence of Opisthorchis viverrini infection and incidence of cholangiocarcinoma in Khon Kaen, Northeast Thailand. Trop Med Int Health. 2004; 9(5):588-94. DOI: 10.1111/j.1365-3156.2004.01234.x. View

15.

Chaiyadet S, Krueajampa W, Hipkaeo W, Plosan Y, Piratae S, Sotillo J . Suppression of mRNAs encoding CD63 family tetraspanins from the carcinogenic liver fluke Opisthorchis viverrini results in distinct tegument phenotypes. Sci Rep. 2017; 7(1):14342. PMC: 5662742. DOI: 10.1038/s41598-017-13527-5. View

16.

Kalra H, Adda C, Liem M, Ang C, Mechler A, Simpson R . Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma. Proteomics. 2013; 13(22):3354-64. DOI: 10.1002/pmic.201300282. View

17.

Shears R, Bancroft A, Hughes G, Grencis R, Thornton D . Extracellular vesicles induce protective immunity against Trichuris muris. Parasite Immunol. 2018; 40(7):e12536. PMC: 6055854. DOI: 10.1111/pim.12536. View

18.

Coakley G, McCaskill J, Borger J, Simbari F, Robertson E, Millar M . Extracellular Vesicles from a Helminth Parasite Suppress Macrophage Activation and Constitute an Effective Vaccine for Protective Immunity. Cell Rep. 2017; 19(8):1545-1557. PMC: 5457486. DOI: 10.1016/j.celrep.2017.05.001. View

19.

Suttiprapa S, Sotillo J, Smout M, Suyapoh W, Chaiyadet S, Tripathi T . Opisthorchis viverrini Proteome and Host-Parasite Interactions. Adv Parasitol. 2018; 102:45-72. DOI: 10.1016/bs.apar.2018.06.002. View

20.

Sotillo J, Pearson M, Potriquet J, Becker L, Pickering D, Mulvenna J . Extracellular vesicles secreted by Schistosoma mansoni contain protein vaccine candidates. Int J Parasitol. 2015; 46(1):1-5. DOI: 10.1016/j.ijpara.2015.09.002. View