Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.5.1.18 (glutathione S-transferase)
 22,582 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Vaccination with a membrane-bound thiol Sepharose-binding fraction (TSBP) of adult Haemonchus contortus has been shown to confer significant levels of protection against homologous challenge in sheep. This fraction is greatly enriched for cysteine proteinase activity. Following fractionation of TSBP by anion-exchange chromatography on MonoQ, protection was found to partition with those fractions further enriched for cysteine proteinase activity. In this study, the cysteine proteinases of adult H. contortus TSBP were specifically purified by affinity chromatography using recombinant H. contortus cystatin, a potent cysteine proteinase inhibitor. Although only 1-1.5% of total TSBP bound to cystatin-Sepharose, this fraction contained 100% of the cysteine proteinase activity, as determined by gelatin substrate gel analysis. When used to immunise sheep, less than 3microg per dose of this cysteine proteinase fraction was found to confer a substantial and repeatable level of protection against homologous challenge infection, reducing faecal egg counts by 48 and 28% and worm burdens by 44 and 46% over two trials. Host serum immunoglobulin levels and abomasal mast cell and eosinophil numbers were evaluated, although no correlation with protection was observed. Three cathepsin B-like cysteine proteinases present in TSBP (hmcp1, 4 and 6) have been identified previously by cDNA library immunoscreening. The predicted mature forms of these three cysteine proteinases were expressed in bacteria as insoluble, GST-fusion proteins. Following solubilisation in urea/DTT, the protective capacity of a cocktail of recombinant proteins was evaluated in sheep. Although no reduction in faecal egg counts was observed, sheep vaccinated with recombinant cysteine proteinases showed a highly significant 38% reduction (P <0.01) in worm burdens.
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PMID:Protection studies in sheep using affinity-purified and recombinant cysteine proteinases of adult Haemonchus contortus. 1547 16

Significant levels of protection against Haemonchus contortus have been achieved in sheep by vaccination with a cysteine proteinase-enriched fraction (TSBP) isolated from the gut of adult parasites. Protection is associated with three cathepsin B-like cysteine proteinases (hmcp 1, 4 & 6). Lambs vaccinated with these proteinases, expressed in bacteria as glutathione S-transferase fusion proteins, had significantly reduced (38%) worm burdens compared to challenge controls although, intriguingly, egg output was unaffected. Here, a repeat trial with similar results is reported and protection obtained compared to that induced by vaccination with the predicted mature forms of hmcp1, 4 and 6 expressed in bacteria as non-fusion proteins. Sheep immunized with a cocktail of these non-fusion proteins had reduced faecal egg counts of 27% (P = 0.17) and worm burdens of 29% (P = 0.01) compared to controls. High levels of host serum IgG were detected in GST-hmcp and non-fusion hmcp-immunized animals, although no correlation with protection could be determined. Sera from these groups bound to the microvillar surface of the gut of H. contortus.
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PMID:Further protection studies using recombinant forms of Haemonchus contortus cysteine proteinases. 1662 7