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Vet Parasitol ; 233: 115-122, 2017 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-28043381

RESUMEN

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.


Asunto(s)
Fasciola/genética , Fascioliasis/inmunología , Fascioliasis/prevención & control , Proteínas del Helminto/inmunología , Proteínas Recombinantes/inmunología , Vacunas Sintéticas/inmunología , Alanina Transaminasa/sangre , Animales , Anticuerpos Antihelmínticos/sangre , Aspartato Aminotransferasas/sangre , Escherichia coli/genética , Fasciola/inmunología , Fascioliasis/parasitología , Proteínas del Helminto/genética , Hígado/patología , Ratones , Ratones Endogámicos ICR , Proteínas Recombinantes/genética
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