title: 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.
authors.affiliation: Department of Pathobiology, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok, 10400, Thailand.
authors.forename: Sirima
authors.initials: S
authors.lastname: Wongwairot
authors.forename: Pornanan
authors.initials: P
authors.lastname: Kueakhai
authors.forename: Narin
authors.initials: N
authors.lastname: Changklungmoa
authors.forename: Wipaphorn
authors.initials: W
authors.lastname: Jaikua
authors.forename: Veerawat
authors.initials: V
authors.lastname: Sansri
authors.forename: Krai
authors.initials: K
authors.lastname: Meemon
authors.forename: Sineenart
authors.initials: S
authors.lastname: Songkoomkrong
authors.forename: Suda
authors.initials: S
authors.lastname: Riengrojpitak
authors.forename: Prasert
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D000293:Adolescent; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D056668:Cathepsin L / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005209:Fasciola / Q000276:immunology / Q000502:physiology*; D005211:Fascioliasis / Q000469:parasitology; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology*; D007159:Immunologic Tests; D058488:Metacercariae; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D056668:Cathepsin L