Monoclonal antibody against recombinant Fasciola gigantica cathepsin L1H could detect juvenile and adult cathepsin Ls of Fasciola gigantica

Abstract

Cathepsin Ls (CatLs), the major cysteine protease secreted by Fasciola spp., are important for parasite digestion and tissue invasion. Fasciola gigantica cathepsin L1H (FgCatL1H) is the isotype expressed in the early stages for migration and invasion. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1H (rFgCatL1H) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with recombinant proFgCatL1H (rproFgCatL1H). This MoAb is an immunoglobulin (Ig)G1 with κ light chain isotype. The MoAb reacted specifically with rproFgCatL1H, the native FgCatL1H at a molecular weight (MW) 38 to 48 kDa in the extract of whole body (WB) of metacercariae and newly excysted juvenile (NEJ) and cross-reacted with rFgCatL1 and native FgCatLs at MW 25 to 28 kDa in WB of 2- and 4-week-old juveniles, adult, and adult excretory-secretory (ES) fractions by immunoblotting and indirect ELISA. It did not cross-react with antigens in WB fractions from other parasites, including Gigantocotyle explanatum, Paramphistomum cervi, Gastrothylax crumenifer, Eurytrema pancreaticum, Setaria labiato-papillosa, and Fischoederius cobboldi. By immunolocalization, MoAb against rFgCatL1H reacted with the native protein in the gut of metacercariae and NEJ and also cross-reacted with CatL1 in 2- and 4-week-old juveniles and adult F. gigantica. Therefore, FgCatL1H and its MoAb may be used for immunodiagnosis of both early and late fasciolosis in ruminants and humans.

Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease

“…gigantica ; however, the MoAb showed strong cross-reactions with other parasite antigens. In addition, Wongwairot et al [ 33 ] revealed that MoAb could detect isoforms of cathepsin Ls of both juvenile and adult F . gigantica .…”

Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection ofFasciola giganticacirculating fatty acid binding protein

“…It was found that cross-reactivity with S. mansoni antigen occurred by using these MoAbs. Another study of F. gigantica revealed that MoAb against isoforms of cathepsin Ls of juvenile and adult fluke was used for diagnosis of animal fasciolosis (Wongwairot et al 2015).…”

Juvenile-specific cathepsin proteases in Fasciola spp.: their characteristics and vaccine efficacies

“…The cross-reactivities among antibodies to cathepsin L isotypes were due to the similarity and identity of their amino acid sequences. Localization of FgCatL1H by both polyclonal and monoclonal antibody showed its expression in the epithelial cells of intestinal tract of metacercariae, NEJ, 2-and 4-week-old juveniles, and adult F. gigantica Wongwairot et al 2015). The positive result in the adult might be due to a high degree of cross-reaction between FgCatL1H, which is the juvenile isoform, with the adult-specific cathepsin L. By contrast, FhCL3 was also detected by using its corresponding antibody in oral and ventral suckers and tegument of F. hepatica NEJ (Zawistowska-Deniziak et al 2013).…”