Contortin

Contortin was the first intestinal antigen of the sheep parasite Haemonchus contortus which induced significant levels of protection when used to vaccinate lambs. This antigen is present in the intestine of L4 and adult worms as a helical polymeric structure attached to the luminal surface of the intestinal cells. However, the nature of the protein itself and its function have never been reported. In the present study, contortin was isolated and analysed by peptide mass fingerprint and LC/MS-MS. These analyses indicated that contortin comprises two major proteins, Hc-PCP1 and Hc-PCP2, with homology to prolyl-carboxypeptidases. The two proteins show 64% amino acid sequence identity to each other and both are comprised of two prolyl-carboxypeptidase S28 type domains organised in a tandem repeat. The transcripts of both genes are present from the L4 stage onwards, coinciding with the onset of blood-feeding. Addition of contortin to a fibrinogen solution significantly inhibited blood coagulation in a dose-dependent manner. Mass-spectrometry indicated that the contortin-enriched fraction degraded the C-terminal end of the fibrinogen alpha-chain, which was shown previously to be essential for clot formation. The process happens within seconds after addition and can be inhibited by the dipeptidyl-peptidase IV inhibitors Diprotin A and Bt-PEG-Glu-Pro(P)(OPh)2. These data suggest that the prolyl-carboxypeptidases are intestinal anticoagulants used by H. contortus to interfere with blood coagulation.

An extract of adult Haemonchus contortus enriched in the parasite's intestinal microvillar membrane protein H11 and other integral membrane proteins but free of the protein contortin was evaluated as a potential vaccine in two breeds of sheep. The worm burdens of Clun Forest sheep injected with the extract and challenged with 25,000 infective larvae were reduced 89% by weight compared to the average for the controls. The worm burdens of Dorset sheep (challenged with 10,000 infective larvae) were reduced 72%. In both breeds the reduction in the number of female worms, 92 and 71.8%, respectively, was greater than the reduction in the males (86.5 and 46%). Parasite egg output, determined only for the Dorsets, was reduced 92% protection correlated with serum antibody titre. Most of the antibodies were directed against H11.

To date, proteins isolated from the surface of the gut of gastrointestinal nematodes, particularly Haemonchus contortus, have generally proved to be useful protective antigens and several are being progressed towards recombinant protein-based vaccines. This paper describes the properties of some of the most promising antigens and summarises their performance in laboratory and field based trials. The antigens described include contortin, H11, H-gal-GP, GPI and cysteine proteinases. In addition, the discussion addresses the utility of selected antigens to protect against co-infecting nematode species such as Teladorsagia circumcincta and against related nematode infections such as Ostertagia ostertagi in cattle.