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Parasitology Research

, Volume 118, Issue 12, pp 3419–3427 | Cite as

Recombinant Opisthorchis viverrini tetraspanin expressed in Pichia pastoris as a potential vaccine candidate for opisthorchiasis

  • Luyen Thi Phung
  • Sujittra Chaiyadet
  • Nuttanan Hongsrichan
  • Javier Sotillo
  • Hang Dinh Thi Dieu
  • Canh Quang Tran
  • Paul J Brindley
  • Alex Loukas
  • Thewarach LahaEmail author
Helminthology - Original Paper
  • 95 Downloads

Abstract

Opisthorchiasis affects millions of people in Southeast Asia and has been strongly associated with bile duct cancer. Current strategic control approaches such as chemotherapy and health education are not sustainable, and a prophylactic vaccine would be a major advance in the prevention of the disease. Tetraspanins are transmembrane proteins previously described as potential vaccine candidates for other helminth infections and are also found in the membranes of the tegument and extracellular vesicles of O. viverrini. Here, we investigated the potential of a recombinant protein encoding for the large extracellular loop of O. viverrini tetraspanin-2 (rOv-LEL-TSP-2) in a hamster vaccination model. Hamsters were vaccinated with 50 and 100 μg of rOv-LEL-TSP-2 produced from Pichia pastoris yeast combined with alum CpG adjuvant via the intraperitoneal route. The number of worms recovered from hamsters vaccinated with rOv-LEL-TSP-2 was significantly reduced compared to adjuvant control groups. Fecal egg output was also significantly reduced in vaccinated animals, and the average length of worms recovered from vaccinated animals was significantly shorter than that of the control group. Vaccinated animals showed significantly increased levels of anti-rOv-TSP-2 IgG in the sera after three immunizations, as well as increased levels of several T helper type 1 cytokines in the spleen including IFN-γ and IL-6 but not the Th2/regulatory cytokines IL-4 or IL-10. These results suggest that rOv-TSP-2 could be a potential vaccine against opisthorchiasis and warrants further exploration.

Keywords

Opisthorchis viverrini Tetraspanin Vaccination Pichia pastoris 

Notes

Funding information

This research was supported by Faculty of Medicine, Khon Kaen University, a project grant from the National Health and Medical Research Council of Australia (NHMRC), grant identification number APP1085309, and the National Cancer Institute, National Institute of Health, grant number 2R01CA164719-06A1. AL is supported by a senior principal research fellowship APP1117504.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

436_2019_6488_Fig5_ESM.png (1006 kb)
Supplemental Fig. 1

Recombinant form of Ov-LEL-TSP-2 expressed in P. pastoris. SDS-PAGE and Western blot analysis of samples from expression and purification of rOv-LEL-TSP-2 protein in P. pastoris. Protein marker (lane M), pre-induction supernatant (lane 1), methanol-induction supernatant (lane 2), flow through (lane 3), wash (lane 4), eluted protein (lane 5), purified protein in 20 mM HEPES (lane 6), western blot of rOv-LEL-TSP-2 probed with rabbit anti-Ov-TSP-2 serum (lane 7), western blot of 20 mM HEPES buffer probed with rabbit anti-Ov-TSP-2 serum-negative control (lane 8). The molecular weight of rOv-LEL-TSP-2 protein is approximately 12 kDa. (PNG 1006 kb)

436_2019_6488_MOESM1_ESM.tif (3.7 mb)
High Resolution (TIF 3770 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Luyen Thi Phung
    • 1
    • 2
  • Sujittra Chaiyadet
    • 1
  • Nuttanan Hongsrichan
    • 1
  • Javier Sotillo
    • 3
    • 4
  • Hang Dinh Thi Dieu
    • 2
  • Canh Quang Tran
    • 2
  • Paul J Brindley
    • 5
  • Alex Loukas
    • 4
  • Thewarach Laha
    • 1
    Email author
  1. 1.Department of Parasitology, Faculty of MedicineKhon Kaen UniversityKhon KaenThailand
  2. 2.Hai Duong Medical Technical UniversityHai Duong CityVietnam
  3. 3.Centro Nacional de MicrobiologíaInstituto de Salud Carlos IIIMadridSpain
  4. 4.Centre for Molecular Therapeutics, Australian Institute of Tropical Health and MedicineJames Cook UniversityCairnsAustralia
  5. 5.Department of Microbiology, Immunology and Tropical MedicineGeorge Washington UniversityWashingtonUSA

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