title: Characterization and vaccine potential of Fasciola gigantica saposin-like protein 1 (SAP-1).
abstract: The recombinant Fasciola gigantica Saposin-like protien-1 (rFgSAP-1) was cloned by polymerase chain reaction (PCR) from NEJ cDNA, expressed in Escherichia coli BL21 (DE3) and used for production of a polyclonal antibody in rabbits (anti-rFgSAP-1). By immunoblotting and immunohistochemistry, rabbit IgG anti-rFgSAP-1 reacted with rFgSAP-1 at a molecular weight 12kDa, but not with rFgSAP-2. The rFgSAP-1 reacted with antisera from mouse infected with F. gigantica metacercariae collected at 2, 4, and 6 weeks after infection. The FgSAP-1 protein was expressed at a high level in the caecal epithelium of metacercariae and NEJs. The vaccination was performed in Imprinting Control Region (ICR) mice (n=10) by subcutaneous injection with 50μg of rFgSAP-1 combined with Alum adjuvant. Two weeks after the second boost, mice were infected with 15 metacercariae per mouse by the oral route. The percents protection of rFgSAP-1 vaccine were estimated to be 73.2% and 74.3% when compared with non vaccinated-infected and adjuvant-infected controls, respectively. The levels of IgG1 and IgG2a specific to rFgSAP-1 in the immune sera, which are indicative of Th2 and Th1 immune responses, were inversely and significantly correlated with the numbers of worm recoveries. The rFgSAP-1-vaccinated mice showed significantly reduced levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and liver damage. These indicated that rFgSAP-1 has strong potential as a vaccine candidate against F. gigantica, whose efficacy will be studied further in large economic animals including cattle, sheep, and goat.
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand.
authors.forename: Pornanan
authors.initials: P
authors.lastname: Kueakhai
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand.
authors.forename: Narin
authors.initials: N
authors.lastname: Changklungmoa
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand.
authors.forename: Pinkamon
authors.initials: P
authors.lastname: Waseewiwat
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand.
authors.forename: Thanaporn
authors.initials: T
authors.lastname: Thanasinpaiboon
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand.
authors.forename: Werachon
authors.initials: W
authors.lastname: Cheukamud
authors.affiliation: Faculty of Veterinary Medicine, Mahanakorn University of Technology, Cheum-Sampan Road, Nong Chok, Bangkok 10530, Thailand.
authors.forename: Pannigan
authors.initials: P
authors.lastname: Chaichanasak
authors.affiliation: Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Mueang District, Chonburi 20131, Thailand; Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand. Electronic address: prasert.sob@mahidol.ac.th.
authors.forename: Prasert
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D001219:Aspartate Aminotransferases / Q000097:blood; D004926:Escherichia coli / Q000235:genetics; D005209:Fasciola / Q000235:genetics* / Q000276:immunology; D005211:Fascioliasis / Q000276:immunology* / Q000469:parasitology / Q000517:prevention & control*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology*; D008099:Liver / Q000473:pathology; D051379:Mice; D008813:Mice, Inbred ICR; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; C406725:FHAP protein, Fasciola hepatica; D015801:Helminth Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase
keywords: Characterization; Fasciola gigantica; Localization; Saposin-like protein 1; Vaccine