Skip to main content
SpringerLink
Log in

Development of Experimental Vaccines Against Liver Flukes

  • Protocol
  • First Online:
Vaccine Design

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1404))

Abstract

A multitude of experimental vaccines have been developed against liver flukes in the past. However, there has yet to be the development of a commercial livestock vaccine. Reasons for this may be multiple, and include the lack of identification of the best antigen(s), or the immune response induced by those antigens not being appropriate in either magnitude or polarity (and therefore not protective). Cathepsin proteases are the major component of the excretory/secretory (ES) material of liver flukes in all stages of their life cycle in the definitive host and are the primary antigens of interest for the vaccine development in many studies. Hence, this chapter presents the methodologies of using cathepsin proteases as targeted antigens in recombinant protein and DNA vaccine development to engender protective immune responses against fasciolosis.

First, the experimental vaccines developed in the past and the criteria of an effective vaccine for fasciolosis are briefly reviewed. Then flowcharts for recombinant protein vaccine and DNA vaccine development are presented, followed by the detailed materials and methodologies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Spithill T, Smooker P, Copeman B (1999) Fasciola gigantica: epidemiology, control, immunology and molecular biology. In: Dalton JP (ed) Fasciolosis. CAB International, Wallingford, UK, pp 465–525

    Google Scholar 

  2. Rim HJ, Farag HF, Sornmani S, Cross JH (1994) Food-borne trematodes: ignored or emerging? Parasitol Today 10(6):207–209

    Google Scholar 

  3. Mas-Coma MS, Esteban JG, Bargues MD (1999) Epidemiology of human fascioliasis: a review and proposed new classification. Bull World Health Organ 77:340–346

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Fairweather I (2009) Triclabendazole progress report, 2005–2009: an advancement of learning? J Helminthol 83:139–150

    CAS  PubMed  Google Scholar 

  5. Gordon D, Zadoks R, Skuce P, Sargison N (2012) Confirmation of triclabendazole resistance in liver fluke in the UK. Vet Rec 171:159–160

    PubMed  Google Scholar 

  6. Brockwell YM, Elliott TP, Anderson GR, Stanton R, Spithill TW, Sangster NC (2014) Confirmation of Fasciola hepatica resistant to triclabendazole in naturally infected Australian beef and dairy cattle. Int J Parasitol Drugs Drug Resist 4:48–54

    PubMed  Google Scholar 

  7. Allen JE, Maizels RM (2011) Diversity and dialogue in immunity to helminths. Nat Rev Immunol 11:375–388

    CAS  PubMed  Google Scholar 

  8. Allen JE, Wynn TA (2011) Evolution of Th2 immunity: a rapid repair response to tissue destructive pathogens. PLoS Pathog 7(5):e1002003

    CAS  PubMed  PubMed Central  Google Scholar 

  9. James SL, Glaven J (1989) Macrophage cytotoxicity against schistosomula of Schistosoma mansoni involves arginine-dependent production of reactive nitrogen intermediates. J Immunol 143:4208–4212

    CAS  PubMed  Google Scholar 

  10. Pulendran B, Smith JL, Caspary G, Brasel K et al (1999) Distinct dendritic cell subsets differentially regulate the class of immune response in vivo. Proc Natl Acad Sci U S A 96:1036–1041

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Wiedosari E, Hayakawa H, Copeman B (2006) Host differences in response to trickle infection with Fasciola gigantica in buffalo, Ongole and Bali calves. Trop Anim Health Prod 38:43–53

    CAS  PubMed  Google Scholar 

  12. Raadsma HW, Kingsford NM, Suharyanta, Spithill TW, Piedrafita D (2007) Host responses during experimental infection with Fasciola gigantica or Fasciola hepatica in Merino sheep: I. Comparative immunological and plasma biochemical changes during early infection. Vet Parasitol 143:275–286

    CAS  PubMed  Google Scholar 

  13. Hansen DS, Clery DG, Estuningsih SE, Widjajanti S, Partoutomo S, Spithill TW (1999) Immune responses in Indonesian thin tail and Merino sheep during a primary infection with Fasciola gigantica: lack of a specific IgG2 antibody response is associated with increased resistance to infection in Indonesian sheep. Int J Parasitol 29:1027–1035

    CAS  PubMed  Google Scholar 

  14. Acosta D, Cancela M, Piacenza L, Roche L, Carmona C, Tort JF (2008) Fasciola hepatica leucine aminopeptidase, a promising candidate for vaccination against ruminant fasciolosis. Mol Biochem Parasitol 158:52–64

    CAS  PubMed  Google Scholar 

  15. Sexton JL, Wilce MC, Colin T, Wijffels GL et al (1994) Vaccination of sheep against Fasciola hepatica with glutathione S-transferase. Identification and mapping of antibody epitopes on a three-dimensional model of the antigen. J Immunol 152:1861–1872

    CAS  PubMed  Google Scholar 

  16. Kennedy NJ, Spithill TW, Tennent J, Wood PR, Piedrafita D (2006) DNA vaccines in sheep: CTLA-4 mediated targeting and CpG motifs enhance immunogenicity in a DNA prime/protein boost strategy. Vaccine 24:970–979

    CAS  PubMed  Google Scholar 

  17. Jayaraj R, Piedrafita D, Dynon K, Grams R, Spithill TW, Smooker PM (2010) Liver fluke vaccines: vaccination against fasciolosis by a multivalent vaccine of recombinant stage-specific antigens. Proc Vaccinol 2:82–85

    CAS  Google Scholar 

  18. Jayaraj R, Piedrafita D, Spithill T, Smooker P (2012) Evaluation of the immune responses induced by four targeted DNA vaccines encoding the juvenile liver fluke antigen, cathepsin B in a mouse model. Genet Vaccines Ther 10:7

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Piacenza L, Acosta D, Basmadjian I, Dalton JP, Carmona C (1999) Vaccination with cathepsin L proteinases and with leucine aminopeptidase induces high levels of protection against fascioliasis in sheep. Infect Immun 67:1954–1961

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Maggioli G, Acosta D, Silveira F, Rossi S et al (2011) The recombinant gut-associated M17 leucine aminopeptidase in combination with different adjuvants confers a high level of protection against Fasciola hepatica infection in sheep. Vaccine 29:9057–9063

    CAS  PubMed  Google Scholar 

  21. Morphew RM, Wright HA, LaCourse EJ, Woods DJ, Brophy PM (2007) Comparative proteomics of excretory-secretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host. Mol Cell Proteomics 6:963–972

    CAS  PubMed  Google Scholar 

  22. Robinson MW, Tort JF, Lowther J, Donnelly SM et al (2008) Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica: expansion of a repertoire of virulence-associated factors. Mol Cell Proteomics 7:1111–1123

    CAS  PubMed  Google Scholar 

  23. Dalton JP, McGonigle S, Rolph TP, Andrews SJ (1996) Induction of protective immunity in cattle against infection with Fasciola hepatica by vaccination with cathepsin L proteinases and with hemoglobin. Infect Immun 64:5066–5074

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Dalton JP, Neill SO, Stack C, Collins P et al (2003) Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines. Int J Parasitol 33:1173–1181

    CAS  PubMed  Google Scholar 

  25. Norbury LJ, Beckham S, Pike RN, Grams R et al (2011) Adult and juvenile Fasciola cathepsin L proteases: different enzymes for different roles. Biochimie 93:604–611

    CAS  PubMed  Google Scholar 

  26. Norbury LJ, Hung A, Beckham S, Pike RN, Spithill TW, Craik CS, Choe Y, Fecondo JV, Smooker PM (2012) Analysis of Fasciola cathepsin L5 by S2 subsite substitutions and determination of the P1–P4 specificity reveals an unusual preference. Biochimie 94:1119–1127

    CAS  PubMed  Google Scholar 

  27. Lecaille F, Choe Y, Brandt W, Li Z, Craik CS, Bromme D (2002) Selective inhibition of the collagenolytic activity of human cathepsin K by altering its S2 subsite specificity. Biochemistry 41:8447–8454

    CAS  PubMed  Google Scholar 

  28. Turk D, Guncar G, Podobnik M, Turk B (1998) Revised definition of substrate binding sites of papain-like cysteine proteases. Biol Chem 379:137–147

    CAS  PubMed  Google Scholar 

  29. Smooker PM, Steeper KR, Drew DR, Strugnell RA, Spithill TW (1999) Humoral responses in mice following vaccination with DNA encoding glutathione S-transferase of Fasciola hepatica: effects of mode of vaccination and the cellular compartment of antigen expression. Parasite Immunol 21:357–364

    CAS  PubMed  Google Scholar 

  30. Mattanovich D, Branduardi P, Dato L, Gasser B et al (2012) Recombinant protein production in yeasts. Methods Mol Biol 824:329–358

    CAS  PubMed  Google Scholar 

  31. Kutzler MA, Weiner DB (2008) DNA vaccines: ready for prime time? Nat Rev Genet 9:776–788

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Babiuk LA, Pontarollo R, Babiuk S et al (2003) Induction of immune responses by DNA vaccines in large animals. Vaccine 21:649–658

    CAS  PubMed  Google Scholar 

  33. Leung JY, Pijlman GP, Kondratieva N, Hyde J et al (2008) Role of nonstructural protein NS2A in flavivirus assembly. J Virol 82:4731–4741

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Mutwiri G, Pontarollo R, Babiuk S, Griebel P et al (2003) Biological activity of immunostimulatory CpG DNA motifs in domestic animals. Vet Immunol Immunopathol 91:89–103

    CAS  PubMed  Google Scholar 

  35. Klinman DM, Currie D, Gursel I, Verthelyi D (2004) Use of CpG oligodeoxynucleotides as immune adjuvants. Immunol Rev 199:201–216

    CAS  PubMed  Google Scholar 

  36. Estuningsih SE, Smooker PM, Wiedosari E, Widjajanti S et al (1997) Evaluation of antigens of Fasciola gigantica as vaccines against tropical fasciolosis in cattle. Int J Parasitol 27:1419–1428

    CAS  PubMed  Google Scholar 

  37. Jayaraj R, Piedrafita D, Dynon K, Grams R, Spithill TW, Smooker PM (2009) Vaccination against fasciolosis by a multivalent vaccine of stage-specific antigens. Vet Parasitol 160:230–236

    CAS  PubMed  Google Scholar 

  38. Kjeldsen T, Brandt J, Andersen AS, Egel-Mitani M et al (1996) A removable spacer peptide in an alpha-factor-leader/insulin precursor fusion protein improves processing and concomitant yield of the insulin precursor in Saccharomyces cerevisiae. Gene 170:107–112

    CAS  PubMed  Google Scholar 

  39. Lesage G, Prat A, Lacombe J, Thomas DY et al (2000) The Kex2p proregion is essential for the biosynthesis of an active enzyme and requires a C-terminal basic residue for its function. Mol Biol Cell 11:1947–1957

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Cui Z (2005) DNA vaccine. In: Leaf Huang M-CH, Ernst W (eds) Advances in genetics, vol 54. Academic, San Diego, CA, pp 257–289

    Google Scholar 

  41. Hemmi H, Takeuchi O, Kawai T, Kaisho T et al (2000) A Toll-like receptor recognizes bacterial DNA. Nature 408:740–745

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Drs Luke Norbury and Rama Jayaraj for undertaking much of the work described here.

Author information

Authors and Affiliations

  1. School of Applied Sciences, RMIT University, Plenty Road, Bundoora, VIC, 3083, Australia

    Huan Yong Yap & Peter M. Smooker

Authors
  1. Huan Yong Yap
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter M. Smooker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Peter M. Smooker .

Editor information

Editors and Affiliations

  1. Lankenau Institute for Medical Research, Wynnewood, PA, USA

    Sunil Thomas

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Yap, H.Y., Smooker, P.M. (2016). Development of Experimental Vaccines Against Liver Flukes. In: Thomas, S. (eds) Vaccine Design. Methods in Molecular Biology, vol 1404. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-3389-1_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3389-1_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-3388-4

  • Online ISBN: 978-1-4939-3389-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation