Enzymatic characteristics and preventive effect of leucine aminopeptidase against Echinococcus multilocularis
•The bioinformatics and enzymatic feature analysis indicated that rEm-LAP belongs to the M17 LAP family and that the soluble protein possesses good activity.•The optimal activity of rEm-LAP is pH 9.•BALB/c mice vaccinated with rEm-LAP had significantly reduced both the number and size of the cysts c...
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Veröffentlicht in: | Acta tropica 2021-10, Vol.222, p.106066-106066, Article 106066 |
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Sprache: | eng |
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Zusammenfassung: | •The bioinformatics and enzymatic feature analysis indicated that rEm-LAP belongs to the M17 LAP family and that the soluble protein possesses good activity.•The optimal activity of rEm-LAP is pH 9.•BALB/c mice vaccinated with rEm-LAP had significantly reduced both the number and size of the cysts compared to control animals that received PBS.•Studies revealed that the immune protection of vaccination with rEm-LAP is likely mediated by a Th1-Th2 mixed reaction.
Alveolar echinococcosis, a parasitic zoonotic disease caused by the larval stage of Echinococcus multilocularis infection, is a global epidemic in Eurasia and North America. Leucine aminopeptidase (LAP) of the M17 peptidase family could act on an ideal target antigen in diagnosis and prevention of parasitic diseases (schistosomiasis, malaria, fascioliasis) because of its good immunogenicity. In this study, the bioinformatic and enzymatic characterizations of recombinant Echinococcus multilocularis LAP (rEm-LAP) were evaluated. A prokaryotic expression system for rEm-LAP protein was established and its immunogenicity and preventive efficacy were demonstrated in a BALB/c mice model. This is the first report about the LAP of Echinococcus multilocularis and with a 57.4 KD purified rEm-LAP protein successfully expressed by pCzn1-LAP in Escherichia coli BL-21 cells. Enzymatic analysis results showed optimal rEm-LAP activity at pH 9. Serum indirect ELISA demonstrated that rEm-LAP could induce a Th1 and Th2 mixed-type immunological response and produce high levels of IgG, IgG1, IgG2a, IgM, and IgA. Furthermore, serum IFN-γ and IL-4 secretion were increased compared with the control groups. Finally, vaccination with rEm-LAP significantly decreased both the number and size of the cysts in Echinococcus multilocularis metacestode infected mice model. The current study provides evidence that rEm-LAP could be a potential vaccine antigen of Echinococcus multilocularis. |
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ISSN: | 0001-706X 1873-6254 |
DOI: | 10.1016/j.actatropica.2021.106066 |