Serum IgG response of Manchego lambs to infections with Haemonchus contortus and preliminary characterization of adult antigens

Cuquerella, Montserrat; Gómez-Muñoz, María Teresa; Alunda, José María

doi:10.1016/0304-4017(91)90123-d

Cited by 31 publications

(20 citation statements)

References 22 publications

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“…Both adult and third-stage larvae (L3) were obtained as previously described (Cuquerella et al 1991). Briefly, L3 were obtained from aerated cultures (26°C, over 80% relative humidity, 10 days) of fecal material of infected lambs and recovered by baermannization.…”

Section: Parasitesmentioning

confidence: 99%

“…Reduced glutathione (GSH) in Tris pH 9.6 was used as elution buffer (Domínguez-Toraño et al 2000). Unbound ASE was fractionated by electrophoresis in a 15% polyacrylamide gel under denaturing and reducing conditions [sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)] and electrotransferred to Immobilon P polyvinylidene fluoride membranes (Millipore; Cuquerella et al 1991). The p26/23 fraction was detected using immune ovine sera (Domínguez-Toraño et al 2000), excised from the membrane, eluted with 1% Triton X 100/2% SDS in 50 mM Tris-HCl pH 9.5 (Szewczyk and Summers 1992) and concentrated with cold acetone.…”

Section: Isolation and Characterization Of Antigensmentioning

confidence: 99%

“…Nylon membranes (Immobilon P, Millipore) were used for the immunodetections, following the protocol described by Cuquerella et al (1991). In short, electrotransfer was carried out in a buffer containing 25 mM Tris, 192 mM glycine, and 20% methanol at 4°C and kept overnight at 30 V, followed by 90 min at 100 V. Membranes were washed in Tris saline buffer (10 mM Tris-HCl, 0.15 M NaCl, pH 7.3) with 0.05% Tween 20 and blocked with skimmed milk (Molico Sveltesse, Nestlé).…”

Section: Western Blottingmentioning

confidence: 99%

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Isolation and immunolocalization of a putative protective antigen (p26/23) from adult Haemonchus contortus

et al. 2008

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The putative protective antigen, p26/23, from adult Haemonchus contortus was isolated. A soluble extract from adult helminths obtained from the abomasa of hyperinfected (12,000 infective larvae) female Manchego lambs and treated with a mixture of protease inhibitors was subjected to affinity chromatography (hexylgluthatione) to eliminate the enzyme gluthatione S-transferase. The eluate was analyzed by electrophoresis under denaturing and reducing conditions (sodium dodecyl sulfate polyacrylamide gel electrophoresis), electrotransferred to nylon membranes, and assayed by Western blot with sera from immunized lambs. The bands recognized by the lambs' sera corresponding to proteins with a molecular weight of 23-26 kDa (p26/23) were excised, eluted, and separated by reverse-phase chromatography. This allowed the isolation of a single protein, which was expressed in both infective larvae (L3) and the adult stage of the parasite. The first 20 amino acids of the N-terminal end of the purified protein were determined. The partial amino acid sequence revealed a 100% identity with the N terminus of one of the peptides present in the p26/23 immunoprotective fraction previously tested by us against sheep haemonchosis. No significant homology with any reported sequence was found except for the deduced sequence of a hypothetic H.contortus protein (HCC00515). Immunolocalization studies showed that the protein was expressed in the hypodermic chords of the nematode.

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Section: Parasitesmentioning

confidence: 99%

Section: Isolation and Characterization Of Antigensmentioning

confidence: 99%

Section: Western Blottingmentioning

confidence: 99%

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Isolation and immunolocalization of a putative protective antigen (p26/23) from adult Haemonchus contortus

et al. 2008

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“…contortus was investigated without any evidence of protection afforded by previous infection of the U.S. Copyright Clearance Center Code Statement: 0931 -1793/94/4106-0399$10.50/0 animals (CUQUERELLA et al, 1991). Since apparently a rapid loss of resistance exists in the absence of parasitic infection (WAGLAND and DINEEN, 1967) these lambs were reallotted and used t o study the response of Manchego adult sheep when infected and challenged 10 weeks later with the parasite; estimating the faecal egg output, serum antibody response by ELISA and Western blotting.…”

Section: Introductionmentioning

confidence: 99%

Partial Protection of Manchego Sheep against Haemonchus contortus after a 6‐Month Postpriming Period. Preliminary Note

Cuquerella

Gómez-Muñoz

Méndez

et al. 1994

Journal of Veterinary Medicine, Series B

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Twelve to 12-5-month-old Manchego sheep which had received a Haernonchus contortus infection with 2500 infective larvae (L-3) when they were 4-4.5 months old (Al,A2) and a challenge with 5000 L-3 2 months later (Al,A2,Bl,B2), finished in both cases with thiabendazole treatment, were subjected to a homologous infection with 7000 L-3 (A1,BI) or kept as control sheep (A2,B2). Fifty days after the infection the animals were treated with rafoxanide and challenged 18 days later with 14,000 L-3 of the parasite.Wide variations were seen, not observed at the 7000 L-3 infection, after challenge among the animals' faecal egg output. Retrospective analysis of the results obtained showed that the sheep response was apparently dependent on the extent of the contact with the parasite 6 months earlier and the elapsed period between priming and infection. A significant protective response was seen at challenge in the sheep which had received 2500 + 5000 L-3 (A2), suggesting the need of a period between 120 and 180 days without infection to show a strong response.No clear correlation was seen between parasitological results and the serum antibody response estimated by ELISA along the experiment and Western blotting analysis during the challenge. However, the less reactive sheep displayed lower ELISA values and simpler immunorecognition patterns in WB.

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“…Notable variations were found in the elapsed posttreatment time required to observe IFAT reductions and were obvious in some dogs (nos. 7,8,9) very early (2 months posttreatment), whereas other dogs (nos. 1, 2) maintained pretreatment IFAT values for longer periods.…”

Section: Resultsmentioning

confidence: 98%

Value of Western Blotting in the Clinical Follow-Up of Canine Leishmaniasis

et al. 1999

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Specific serum antibody levels in Leishmania infantum-infected dogs treated with a combination of glucantime and allopurinol were estimated by indirect immunofluorescence and Western blotting. The sensitivity of Western blot was greater than that obtained with immunofluorescence titration. In general, both diagnostic methods concurred with the post-treatment clinical status of the animals. Clinical improvement of successfully treated dogs was related to lower immunofluorescence titers and simpler and/or less reactive immunodetection patterns in Western blotting. The recognition, by infected dogs, of certain low molecular weight antigens, particularly one of approximately 26 kDa, was restricted to pretreatment samples and a single animal in relapse thus apparently constituting an active infection marker.

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Serum IgG response of Manchego lambs to infections with Haemonchus contortus and preliminary characterization of adult antigens

Cited by 31 publications

References 22 publications

Isolation and immunolocalization of a putative protective antigen (p26/23) from adult Haemonchus contortus

Isolation and immunolocalization of a putative protective antigen (p26/23) from adult Haemonchus contortus

Partial Protection of Manchego Sheep against Haemonchus contortus after a 6‐Month Postpriming Period. Preliminary Note

Value of Western Blotting in the Clinical Follow-Up of Canine Leishmaniasis

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