Presence of the LDNF glycan on the host-protective H-gal-GP fraction from Haemonchus contortus.
Immunization of sheep with the gut membrane-associated protein complex H-gal-GP of adult Haemonchus contortus induces high levels of protection against a homologous challenge infection. Protection is correlated with a systemic IgG response against the antigen. Analysis of the antibody response showed that the majority of the antigen-specific IgG was of the IgG2 isotype. A substantial proportion (74%) of this response was directed against the glycan component of H-gal-GP. The high immunogenicity of the H-gal-GP glycans may be due to the presence of the fucosylated LacdiNAc (LDNF) antigen. 2D electrophoresis, Western blotting and mass spectrometry analysis of H-gal-GP showed that this glycan epitope was specifically located on a metalloendopeptidase, MEP3. MEP3 is the most abundant protein in H-gal-GP and has been identified as one of the most likely protective components of the complex. Here, we present evidence that the LDNF glycan does not contribute to the protective capacity of H-gal-GP. Animals vaccinated with reduced and denatured H-gal-GP are not protected against subsequent infection, although the antibody response against the LDNF glycan is very similar to that of animals vaccinated with the native H-gal-GP. In addition, an alternative version of H-gal-GP, H-sialgal-GP, which is equally protective, but isolated by affinity chromatography on jacalin lectin rather than peanut lectin, contains a MEP3 component which has no detectable LDNF glycan.
Geldhof
P
P
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, UK. geldp@mri.sari.ac.uk
Newlands
G F J
GF
Nyame
K
K
Cummings
R
R
Smith
W D
WD
Knox
D P
DP
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D010450:Endopeptidases / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D006188:Haemonchiasis / Q000276:immunology* / Q000517:prevention & control; D006189:Haemonchus / Q000276:immunology*; D015801:Helminth Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D007785:Lactose / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D008562:Membrane Glycoproteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012756:Sheep; D014611:Vaccination
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; C488538:N-acetylgalactosaminyl-1-4-(fucosyl-1,3)-N-acetylglucosamine; D010450:Endopeptidases; C406387:H-sialgal-GP protein, Haemonchus contortus; C119206:galactose-containing glycoprotein, Haemonchus contortus; D007785:Lactose