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Characterization and expression of cathepsin B2 in Fasciola gigantica.

Abstract

Fasciola gigantica cathepsin B belongs to a family of cysteine proteases which is involved in invasion of host tissues. In this study, the recombinant cathepsin B2 (rFgCatB2), synthesized in Pichia pastoris, showed enzymatic activity on a fluorometric substrate Z-Phe-Arg-AMC and gelatin. Furthermore, this recombinant enzyme could degrade IgG and type I collagen. Mouse antiserum against rFgCatB2 reacted with the native FgCatB2 in whole body (WB) extracts of metacercariae (MET), newly excysted juveniles (NEJ) and 2week-old juveniles, but not in 3, 4 week-old juveniles and adult flukes. Immunolocalization showed the presence of cathepsin B2 only in the caecal epithelium of MET, NEJ and 2 week-old juveniles. Co-localization of FgCatB2 and a prominent antigen of NEJ, FgCatB3, revealed that these proteins were expressed at the same regions in the caecal epithelium. Anti-rFgCatB2 showed no cross reaction with the other parasites' antigens by Western blotting. These findings suggest that CatB2 is expressed only in early stages of the parasite and may be involved in digestion of host connective tissues and evasion of the host immune system during their penetration and migration. Thus, CatB2 could be considered as an immunodiagnostic and vaccine candidate for fasciolosis.

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  • Publisher Full Text
  • Authors

    Chantree P, Wanichanon C, Phatsara M, Meemon K, Sobhon P

    Institution

    Department of Anatomy, Mahidol University, Bangkok 10400, Thailand.

    Source

    Experimental parasitology 132:2 2012 Oct pg 249-56

    MeSH

    Animals
    Antigens, Helminth
    Blotting, Western
    Cathepsin B
    Cattle
    Cecum
    Cricetinae
    Cross Reactions
    Electrophoresis, Polyacrylamide Gel
    Fasciola
    Fluorescent Antibody Technique
    Gene Expression Regulation, Enzymologic
    Lymnaea
    Mesocricetus
    Metacercariae
    Mice
    Mice, Inbred ICR
    Microscopy, Confocal
    Pichia
    Recombinant Proteins

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    22885402