Fasciola hepatica Cathepsin L Zymogens: Immuno-Proteomic Evidence for Highly Immunogenic Zymogen-Specific Conformational Epitopes to Support Diagnostics Development

J Proteome Res. 2022 Aug 5;21(8):1997-2010. doi: 10.1021/acs.jproteome.2c00299. Epub 2022 Jul 18.

Abstract

Fasciola hepatica, the common liver fluke and causative agent of zoonotic fasciolosis, impacts on food security with global economic losses of over $3.2 BN per annum through deterioration of animal health, productivity losses, and livestock death and is also re-emerging as a foodborne human disease. Cathepsin proteases present a major vaccine and diagnostic target of the F. hepatica excretory/secretory (ES) proteome, but utilization in diagnostics of the highly antigenic zymogen stage of these proteins is surprisingly yet to be fully exploited. Following an immuno-proteomic investigation of recombinant and native procathepsins ((r)FhpCL1), including mass spectrometric analyses (DOI: 10.6019/PXD030293), and using counterpart polyclonal antibodies to a recombinant mutant procathepsin L (anti-rFhΔpCL1), we have confirmed recombinant and native cathepsin L zymogens contain conserved, highly antigenic epitopes that are conformationally dependent. Furthermore, using diagnostic platforms, including pilot serum and fecal antigen capture enzyme-linked immunosorbent assay (ELISA) tests, the diagnostic capacities of cathepsin L zymogens were assessed and validated, offering promising efficacy as markers of infection and for monitoring treatment efficacy.

Keywords: cathepsin; diagnostics; fasciolosis; recombinant; triclabendazole.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cathepsin L / genetics
  • Cathepsin L / metabolism
  • Enzyme Precursors
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes
  • Fasciola hepatica* / chemistry
  • Fasciola hepatica* / genetics
  • Fascioliasis* / diagnosis
  • Humans

Substances

  • Enzyme Precursors
  • Epitopes
  • Cathepsin L