Vaccination of Sheep with Purified Cysteine Proteinases of Fasciola hepatica Decreases Worm Fecundity

Wijffels, Gene; Salvatore, Liliana; Dosen, M; Waddington, Josh; Wilson, Lachlan; Thompson, Cindy; Campbell, Noel J.; Sexton, Jennifer; Wicker, Jason A.; Bowen, FL; Friedel, Thorsten; Spithill, Terry W.

doi:10.1006/expr.1994.1014

Cited by 119 publications

(53 citation statements)

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“…For the deglycosylation experiment, 150 ng HieACE was treated with 20 mU N-glycosidase F (Boehringer) as described [21]. Immunoblots were performed as described [33] with the following changes : the nitrocellulose was incubated for 1 h (37°C) with ovine anti-HieACE serum (1 : 2000) then with the secondary antibody conjugated to horseradish peroxidase ( I : 1000) for 1 h (37°C). Blots were developed with 4-chloro-naphthol or (where indicated) by means of a more-sensitive mixed-chromogen-substrate system [34].…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterisation of Angiotensin‐Converting Enzyme from the Dipteran Species, Haematobia irritans exigua, and Its Expression in the Maturing Male Reproductive System

Wijffels

Fitzgerald

Gough

et al. 1996

European Journal of Biochemistry

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The angiotensin-converting enzymes (ACE) are involved in the regulation of the specific maturation or degradation of a number of mammalian bioactive peptides. A carboxydipeptidase similar to mammalian ACE has now been identified in the adult stage of the haematophagous fly, Haematobia irritans exigua (buffalo fly), a close relative of the horn fly of North America. The enzyme was purified by lectin-affinity chromatography and ion-exchange chromatography and migrated as a doublet of 70 kDa upon reducing SDSPAGE. Unlike mammalian ACE, the fly carboxydipeptidase (HieACE) is not membrane bound. The amino acid sequence of an internal peptide from HieACE and a conserved amino acid region present in all mammalian ACE were used to design degenerate oligonucleotide primers suitable for PCR. A DNA fragment amplified from adult buffalo fly cDNA was used to identify a cDNA clone that encoded the enzyme. The cDNA sequence encodes a carboxydipeptidase with 41-42% amino acid identity to the mammalian testicular ACE. The active-site regions of mammalian ACE are conserved in the deduced amino acid sequence of HieACE. Enzymatically, HieACE is very similar to its mammalian counterparts, with comparable K. , and V,,, values for the synthetic substrate, benzoylglycylglycylglycine, and similar patterns of inhibition by EDTA, ACE inhibitor peptide and captopril. HieACE also specifically activates angiotensin I to angiotensin I1 and degrades other mammalian ACE substrates such as bradykinin, substance P and cholecystokinin-8. In the adult fly, HieACE is expressed in the compound ganglion and in the posterior region of the midgut. Similar to the mammalian system, expression of this enzyme is induced in the maturing male reproductive system, which suggests conservation of ACE function in these species.

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Section: Methodsmentioning

confidence: 99%

Cloning and Characterisation of Angiotensin‐Converting Enzyme from the Dipteran Species, Haematobia irritans exigua, and Its Expression in the Maturing Male Reproductive System

Wijffels

Fitzgerald

Gough

et al. 1996

European Journal of Biochemistry

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“…The function of these enzymes is unknown, but they might enable the parasite to destroy host tissue for feeding and penetration, and to evade the host immune system (Tort et al, 1999). The protective capacity of cysteine proteases has already been demonstrated against Fasciola hepatica (Wijffels et al, 1994;Dalton et al, 1996) and H. contortus (Boisvenue et al, 1992;Knox et al, 1999Knox et al, , 2005Redmond and Knox, 2004).…”

Section: Introductionmentioning

confidence: 99%

Vaccination against Ostertagia ostertagi with subfractions of the protective ES-thiol fraction

Meyvis¹,

Geldhof²,

Gevaert³

et al. 2007

Veterinary Parasitology

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Previous vaccination trials against Ostertagia ostertagi in cattle have demonstrated the protective capacity of a protein fraction termed ES-thiol, which is enriched for activation-associated secreted proteins (ASPs) and cysteine proteases. In this study, ES-thiol was subfractionated through Q-Sepharose anion exchange chromatography to determine whether the ASPs and/or the cysteine proteases are responsible for the induced protection. Calves (seven/group) were immunized three times intramuscularly with 100 mg of ES-thiol or equivalent amounts of an ASP-enriched fraction, a cysteine protease-enriched fraction or a rest fraction, with QuilA adjuvant. A negative control group only received QuilA. After the final immunization the animals were challenged with a trickle infection of 25,000 infectious L3 larvae (1000 L3/day; 5 days/week). During a 2-month period the geometric mean cumulative faecal egg count (FEC) of the ES-thiol group was reduced by 62% compared to the QuilA control group (P < 0.05). Groups injected with the ASP-enriched, the cysteine protease-enriched and the rest fraction demonstrated a reduction in cumulative FEC of 74, 80 and 70%, respectively (P < 0.01). Although no significant reductions in worm burdens were observed, adult male and female worms were significantly smaller in all vaccinated groups (P < 0.05), except for male worms from the ES-thiol group. These results suggest the protective capacity of ASPs and the presence of other protective antigens in the ES-thiol fraction. #

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“…Therefore, me dical prophylaxis could be used to avoid these disadvantages. Several vaccination studies have b een carried out by infecting animais with F a s c io la h e p a t ic a irradiated m etacercariae (Cam pbell et al, 1978) or by immunizing them with parasitic substances, like excretorysecretory products (Rajasekariah et a l., 1979), T1 antigen (Hanna et al, 1988), glutathione S transferase (Sexton et al, 1990), cathepsin-L protease (Wijffels et al, 1994), haem oglobin (Dalton et al, 1996). These have not given com plete protection against F. h e p a tica.…”

Section: Introductionmentioning

confidence: 99%

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

1998

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Vaccination of Sheep with Purified Cysteine Proteinases of Fasciola hepatica Decreases Worm Fecundity

Cited by 119 publications

References 0 publications

Cloning and Characterisation of Angiotensin‐Converting Enzyme from the Dipteran Species, Haematobia irritans exigua, and Its Expression in the Maturing Male Reproductive System

Cloning and Characterisation of Angiotensin‐Converting Enzyme from the Dipteran Species, Haematobia irritans exigua, and Its Expression in the Maturing Male Reproductive System

Vaccination against Ostertagia ostertagi with subfractions of the protective ES-thiol fraction

IFNγ and IL-10 production by hepatic lymph node and peripheral blood lymphocytes inFasciola hepaticainfected sheep

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