Construction and mouse antibody response evaluation of juvenile stage-specific chimeric protein from Fasciola gigantica
Fasciolosis, caused by the liver fluke Fasciola gigantica, is a major parasitic disease that affects livestock and therefore causes significant economic losses in tropical countries. Although anthelminthic drugs can kill the parasite, drug-resistant liver fluke populations are increasing. In this st...
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Veröffentlicht in: | Veterinary parasitology 2024-10, Vol.331, p.110254, Article 110254 |
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Zusammenfassung: | Fasciolosis, caused by the liver fluke Fasciola gigantica, is a major parasitic disease that affects livestock and therefore causes significant economic losses in tropical countries. Although anthelminthic drugs can kill the parasite, drug-resistant liver fluke populations are increasing. In this study, a recombinant F. gigantica chimeric protein (rFgCHI) consisting of cathepsin L1H (FgCL1H), cathepsin B3 (FgCB3), and Saposin-like protein 1 (FgSAP1) was designed and expressed in Escherichia coli (BL21). The molecular weight of rFgCHI was 61 kDa. To study the antibody response, male BALB/c mice were immunized via the subcutaneous injection of rFgCHI combined with Quil A. Immunization with rFgCHI showed the induction of IgG1 and IgG2a with a higher IgG1 isotype level, indicating the potential of mixed Th1/Th2 immune responses, with Th2 predominating. However, the results showed high levels of IgG against the single proteins, except for rFgSAP1. Through Western blotting, mouse anti-rFgCHI polyclonal antibodies could be detected to the native proteins obtained from the parasite at all stages. Immunolocalization also revealed that the anti-rFgCHI antibodies could detect targeted antigens in the cecal epithelium of the parasite. These results demonstrated that rFgCHI is immunogenic to the mouse immune system and may potentially be a protein candidate for the development of a fasciolosis vaccine.
•The first chimeric protein (rFgCHI) with three different proteins was constructed.•Anti-rFgCHI antibody recognized the liver fluke at all stages.•Anti-rFgCHI antibody could detect the native tissue on cecal epithelium.•Cross-reactivity of anti-rFgCHI were not found in the other related parasites.•This study highlighted the key design criteria for chimeric protein production. |
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ISSN: | 0304-4017 1873-2550 1873-2550 |
DOI: | 10.1016/j.vetpar.2024.110254 |