Folia Parasitologica 70:017 (2023) | DOI: 10.14411/fp.2023.017

Tetraspanins from the liver fluke Opisthorchis viverrini stimulate cholangiocyte migration and inflammatory cytokine production

Apisit Ruangsuwast¹, Michael J. Smout ORCID...², Paul J. Brindley ORCID...³, Alex Loukas ORCID...², Thewarach Laha ORCID...¹, Sujittra Chaiyadet ORCID...^{4, *}

¹ Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand;

² Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;

³ Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine & Health Sciences, George Washington University, Washington, D.C., USA;

⁴ Department of Tropical Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand *Address for correspondence: Sujittra Chaiyadet, Department of Tropical Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002 Thailand. E-mail: sujitch@kku.ac.th.

The liver fluke Opisthorchis viverrini (Poirier, 1886) (Digenea) secretes extracellular vesicles (EVs) bearing CD63-like tetraspanins on their surface. Fluke EVs are actively internalised by host cholangiocytes in the bile ducts, where they drive pathology and promote neoplasia through induction of cellular proliferation and secretion of inflammatory cytokines. We investigated the effects of tetraspanins of the CD63 superfamily by co-culturing recombinant forms of the large extracellular loop (LEL) of O. viverrini tetraspanin-2 (rLEL-Ov-TSP-2) and tetraspanin-3 (rLEL-Ov-TSP-3) with non-cancerous human bile duct (H69) and cholangiocarcinoma (CCA, M213) cell lines. The results showed that cell lines co-cultured with excretory/secretory products from adult O. viverrini (Ov-ES) underwent significantly increased cell proliferation at 48 hours but not 24 hours compared to untreated control cells (P < 0.05), whereas rLEL-Ov-TSP-3 co-culture resulted in significantly increased cell proliferation at both 24 hours (P < 0.05) and 48 hours (P < 0.01) time points. In like fashion, H69 cholangiocytes co-cultured with both Ov-ES and rLEL-Ov-TSP-3 underwent significantly elevated Il-6 and Il-8 gene expression for at least one of the time points assessed. Finally, both rLEL-Ov-TSP-2 and rLEL-Ov-TSP-3 significantly enhanced migration of both M213 and H69 cell lines. These findings indicated that O. viverrini CD63 family tetraspanins can promote a cancerous microenvironment by enhancing innate immune responses and migration of biliary epithelial cells.

Keywords: cholangiocarcinoma, host-parasite interaction, CD63-like tetraspanins, extracellular vesicle (EVs)

Received: May 16, 2023; Revised: July 7, 2023; Accepted: July 26, 2023; Published online: September 16, 2023  Show citation