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Vaccine potential of recombinant saposin-like protein 2 against Fasciolosis gigantica in mice
Authors:Pornanan Kueakhai  Narin Changklungmoa  Suda Riengrojpitak  Pannigan Chaichanasak  Krai Meemon  Kulathida Chaithirayanon  Pathanin Chantree  Veerawat Sansri  Tadashi Itagaki  Prasert Sobhon
Affiliation:1. Department of Pathobiology, Faculty of Science, Mahidol University, Rama 6 Road, Ratchathewi, Bangkok 10400, Thailand;2. Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand;3. Faculty of Veterinary Medicine, Mahanakorn University of Technology, Cheum-Sampan Road, Nong Chok, Bangkok 10530, Thailand;4. Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Ratchathewi, Bangkok 10400, Thailand;5. Faculty of Physical Therapy, Mahidol University, Putthamonthon Sai 4 Road, Salaya, Putthamonthon, Nakhon Pathom 73170, Thailand;6. Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka 020-8550, Japan
Abstract:Saposin-like protein 2 (SAP-2) is a protein that adult of Fasciola spp. use to lyse plasma membrane of red blood cells, so that their contents can be digested by proteases for the parasites’ nutrients. Thus SAP-2 is a plausible target for vaccination against these parasites. Recombinant Fasciola gigantica saposin-like protein 2 (rFgSAP-2) was expressed in Escherichia coli BL21 (DE3). A vaccination was performed in ICR mice (n = 10) by subcutaneous injection with 50 μg of rFgSAP-2 combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 30 F. gigantica metacercariae by oral route. The percentages of protection of rFgSAP-2 vaccine against F. gigantica were estimated to be 76.4–78.5% when compared with non vaccinated-infected and adjuvant-infected controls, respectively. The antibodies in immune sera of vaccinated mice were shown by immuno-blotting to react with native FgSAP-2 in the extract of 2- and 4-week-old juvenile parasites. By determining the levels of IgG1 and IgG2a in the immune sera, which are indicative of Th2 and Th1 immune responses, it was found that both Th1 and Th2 humoral immune response were significantly increased in rFgSAP-2 immunized group compared with the control groups, with higher levels of Th2 (IgG1) than Th1 (IgG2a). The levels of serum aspartate aminotransferase (AST) and alanine transaminase (ALT) in rFgSAP-2-immunized group showed no significant difference from those of the non-immunized and infected group, indicating that early juvenile parasites induced liver parenchyma damage, even though the numbers of worm recoveries were significantly different. This study indicates that rFgSAP-2 has a high potential as a vaccine candidate against F. gigantica in mice, and this potential will be tested in larger economic animals.
Keywords:Fasciola gigantica   Saposin-like protein 2   Vaccine   Mice   Protection
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