Proteases from Schistosoma mansoni cercariae cleave IgE at solvent exposed interdomain regions

Mol Immunol. 2008 Jan;45(2):567-74. doi: 10.1016/j.molimm.2007.05.021. Epub 2007 Jul 12.

Abstract

Parasitic infections, including schistosomiasis, are associated with high titres of specific and non-specific IgE antibody, and many reports show an in vitro role for IgE in parasite killing. Despite an active immune response, schistosomes survive for long periods in the human bloodstream, implying that the parasite is able to overcome or evade the IgE response mounted against it. One such mechanism is through cleavage of IgE into non-functional fragments by potent parasite derived enzymes. Using domain swap antibodies, recombinant Fcepsilon, and C-terminally tagged Cepsilon4 domains, we have narrowed down the principal cleavage sites to the Cepsilon2/Cepsilon3 and Cepsilon3/Cepsilon4 interdomain region of the IgE-Fc. Two serine proteases, one chymotrypsin-like and the second trypsin-like, have been proposed to be involved. Inhibition assays using selective inhibitors confirmed that both proteases contribute to Fc cleavage, although the chymotrypsin-like enzyme makes the greater contribution. Protein sequencing of IgE fragments cleaved by highly pure preparations of the chymotrypsin-like enzyme revealed that cleavage also occurred post Lys residues within kappa light chain dimers (LELK/GA). Related sequences are found in myosin, thrombospondin, collagen and actin-related proteins; macromolecules present in the skin and through which cercariae must penetrate to initiate an infection. Chemical knockout experiments using specific inhibitors and chromogenic substrates allowed us to show that the trypsin-like enzyme was responsible for light chain cleavage. The finding that pathogenic proteases can cleave the Fc of IgE may provide a useful biochemical tool for the further analysis of IgE structure. Indeed, the finding may raise new possibilities for treatment of IgE-mediated allergic reactions mediated through Fcepsilon-receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies
  • Bence Jones Protein / metabolism
  • Computational Biology
  • Dimerization
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Immunoglobulin E / chemistry*
  • Immunoglobulin E / metabolism*
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Pancreatic Elastase / isolation & purification
  • Pancreatic Elastase / metabolism*
  • Protein Structure, Tertiary
  • Rats
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Schistosoma mansoni / drug effects
  • Schistosoma mansoni / enzymology*
  • Solvents / metabolism*
  • Substrate Specificity / drug effects

Substances

  • Antibodies
  • Enzyme Inhibitors
  • Recombinant Proteins
  • Solvents
  • Immunoglobulin E
  • Bence Jones Protein
  • Pancreatic Elastase