Cloning and characterization of a beta-galactoside-binding protein (galectin) from the gut of the gastrointestinal nematode parasite Haemonchus contortus

Parasitology. 1999 Nov:119 ( Pt 5):483-90. doi: 10.1017/s003118209900503x.

Abstract

A cDNA encoding a beta-galactoside-binding lectin (galectin) was identified by immunoscreening a Haemonchus contortus cDNA library with antisera from lambs vaccinated with a membrane protein complex (H-gal-GP) derived from the parasites' gut. The cDNA sequence, exhibiting a tandem repeat structure and designated Hco-gal-2, showed significant levels of similarity with galectins from several species of nematode as well as mammalian galectin type 4. Native galectin was preferentially extracted from the H-gal-GP complex and also from an insoluble membrane fraction prepared from adult worms using lactose-agarose affinity chromatography. The affinity-purified material had apparent molecular mass of around 35 kDa with 3 distinct bands visible on SDS-PAGE. All 3 bands were identified as galectins by reaction with antiserum raised to recombinant Hco-GAL-2 on Western blot. To determine whether H. contortus galectins have any protective capacity against infection lambs were vaccinated with affinity-purified galectin and subsequently given a single challenge infection. Vaccination did not confer any protection against infection with H. contortus as judged by faecal egg output or worm counts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Helminth / biosynthesis
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / chemistry
  • DNA, Helminth / chemistry
  • DNA, Helminth / isolation & purification
  • Electrophoresis, Polyacrylamide Gel / veterinary
  • Female
  • Galactosides / chemistry
  • Galectins
  • Gene Expression Regulation, Developmental
  • Gene Library
  • Haemonchiasis / immunology
  • Haemonchiasis / prevention & control
  • Haemonchiasis / veterinary*
  • Haemonchus / chemistry
  • Haemonchus / genetics*
  • Haemonchus / immunology
  • Hemagglutinins / chemistry
  • Hemagglutinins / genetics*
  • Hemagglutinins / immunology
  • Male
  • Molecular Sequence Data
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Sheep
  • Sheep Diseases / immunology
  • Sheep Diseases / parasitology*
  • Sheep Diseases / prevention & control
  • Vaccination / veterinary

Substances

  • Antibodies, Helminth
  • DNA Primers
  • DNA, Helminth
  • Galactosides
  • Galectins
  • Hemagglutinins
  • Recombinant Proteins

Associated data

  • GENBANK/AF036098