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Immunization against Lamb Haemonchosis with a Recombinant Somatic Antigen of Haemonchus contortus (rHcp26/23).

García-Coiradas L, Angulo-Cubillán F, Valladares B, Martínez E, de la Fuente C, Alunda JM, Cuquerella M - Vet Med Int (2010)

Bottom Line: Our previous results showed that a soluble fraction from adult stages of the nematode (p26/23) induced partial protection against challenge.Recombinant DNA technology was applied to obtain a synthetic protein (rHcp26/23).Immunization did not induce any significant protection after challenge with 16000 infective larvae of H. contortus, and comparable values for parasite faecal egg output, packed cell volume, and abomasal parasite burdens were found in vaccinated and control animals.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.

ABSTRACT
Haemonchosis, caused by the abomasal nematode Haemonchus contortus, is a common parasitic disease of sheep. Our previous results showed that a soluble fraction from adult stages of the nematode (p26/23) induced partial protection against challenge. Recombinant DNA technology was applied to obtain a synthetic protein (rHcp26/23). Immunological assays (ELISA, Western blotting, and immunolocalization), using sera from lambs immunized with p26/23, confirmed the identity of the recombinant protein and demonstrated that the synthetic protein is equivalent to the purified protein employed in the previous immunoprophylaxis studies. Vaccination of lambs with 300 mug of rHcp26/23 and Freund's adjuvant elicited a notable specific antibody response. Immunization did not induce any significant protection after challenge with 16000 infective larvae of H. contortus, and comparable values for parasite faecal egg output, packed cell volume, and abomasal parasite burdens were found in vaccinated and control animals.

No MeSH data available.


Related in: MedlinePlus

Serum-specific IgG response, determined by ELISA, of lambs along immunization with rHcp26/23 and challenge with 16000 third-stage larvae of H. contortus, against rHcp26/23 (a) and adult soluble extract of the parasite (ASE). G1 (■): lambs (n = 5) immunized with 3 doses (100 μg each) of recombinant p26/23 (rHcp26/23) purified under nondenaturing conditions and challenged; G2 (♦): lambs vaccinated with 3 doses of rHcp26/23 purified under denaturing conditions and challenged; G3 (▲): lambs receiving adjuvant injections and challenged; G4 (): unvaccinated and challenged lambs. Arrow: day of experimental challenge. Values are means ± standard deviation of three determinations.
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fig4: Serum-specific IgG response, determined by ELISA, of lambs along immunization with rHcp26/23 and challenge with 16000 third-stage larvae of H. contortus, against rHcp26/23 (a) and adult soluble extract of the parasite (ASE). G1 (■): lambs (n = 5) immunized with 3 doses (100 μg each) of recombinant p26/23 (rHcp26/23) purified under nondenaturing conditions and challenged; G2 (♦): lambs vaccinated with 3 doses of rHcp26/23 purified under denaturing conditions and challenged; G3 (▲): lambs receiving adjuvant injections and challenged; G4 (): unvaccinated and challenged lambs. Arrow: day of experimental challenge. Values are means ± standard deviation of three determinations.

Mentions: Immunization with the recombinant protein elicited a strong specific IgG systemic response against the recombinant protein after the first immunizing injection (Figure 4(a)). Interestingly, immunized animals with the recombinant protein also displayed a strong response in ELISA against ASE of H. contortus (Figure 4(b)). Comparable results were observed in the Western blots with individual lambs' sera from the patent period of the infection (not shown). Similarly, immunization induced significantly higher lymphoproliferative response of lambs after challenge against the recombinant protein (Figure 5(a)) and soluble extracts of H. contortus (Figure 5(b)) and larval extracts of the helminth (not shown). Immunized lambs (G1 and G2) showed a slight shortening of the duration of prepatent period (18 days) when compared to nonvaccinated and challenged animals (G3 and G4). However, average data of parasite eggs faecal shedding from immunized animals were higher than those found in the control groups (Figure 6). These differences were not significant given the high variation found within each experimental group. All challenged animals displayed a transient fall of both PCV and haemoglobin concentration (not shown), during the prepatent infection and without differences among groups. Adult helminths were recovered from all challenged animals at necropsy (Table 1). The recovery rate (%) of the infective dose administered was low and no evidence of protection afforded by immunization with rHcp26/23 purified under nondenaturing (Group 1) or denaturing (Group 2) conditions was obtained (P > .5).


Immunization against Lamb Haemonchosis with a Recombinant Somatic Antigen of Haemonchus contortus (rHcp26/23).

García-Coiradas L, Angulo-Cubillán F, Valladares B, Martínez E, de la Fuente C, Alunda JM, Cuquerella M - Vet Med Int (2010)

Serum-specific IgG response, determined by ELISA, of lambs along immunization with rHcp26/23 and challenge with 16000 third-stage larvae of H. contortus, against rHcp26/23 (a) and adult soluble extract of the parasite (ASE). G1 (■): lambs (n = 5) immunized with 3 doses (100 μg each) of recombinant p26/23 (rHcp26/23) purified under nondenaturing conditions and challenged; G2 (♦): lambs vaccinated with 3 doses of rHcp26/23 purified under denaturing conditions and challenged; G3 (▲): lambs receiving adjuvant injections and challenged; G4 (): unvaccinated and challenged lambs. Arrow: day of experimental challenge. Values are means ± standard deviation of three determinations.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2902039&req=5

fig4: Serum-specific IgG response, determined by ELISA, of lambs along immunization with rHcp26/23 and challenge with 16000 third-stage larvae of H. contortus, against rHcp26/23 (a) and adult soluble extract of the parasite (ASE). G1 (■): lambs (n = 5) immunized with 3 doses (100 μg each) of recombinant p26/23 (rHcp26/23) purified under nondenaturing conditions and challenged; G2 (♦): lambs vaccinated with 3 doses of rHcp26/23 purified under denaturing conditions and challenged; G3 (▲): lambs receiving adjuvant injections and challenged; G4 (): unvaccinated and challenged lambs. Arrow: day of experimental challenge. Values are means ± standard deviation of three determinations.
Mentions: Immunization with the recombinant protein elicited a strong specific IgG systemic response against the recombinant protein after the first immunizing injection (Figure 4(a)). Interestingly, immunized animals with the recombinant protein also displayed a strong response in ELISA against ASE of H. contortus (Figure 4(b)). Comparable results were observed in the Western blots with individual lambs' sera from the patent period of the infection (not shown). Similarly, immunization induced significantly higher lymphoproliferative response of lambs after challenge against the recombinant protein (Figure 5(a)) and soluble extracts of H. contortus (Figure 5(b)) and larval extracts of the helminth (not shown). Immunized lambs (G1 and G2) showed a slight shortening of the duration of prepatent period (18 days) when compared to nonvaccinated and challenged animals (G3 and G4). However, average data of parasite eggs faecal shedding from immunized animals were higher than those found in the control groups (Figure 6). These differences were not significant given the high variation found within each experimental group. All challenged animals displayed a transient fall of both PCV and haemoglobin concentration (not shown), during the prepatent infection and without differences among groups. Adult helminths were recovered from all challenged animals at necropsy (Table 1). The recovery rate (%) of the infective dose administered was low and no evidence of protection afforded by immunization with rHcp26/23 purified under nondenaturing (Group 1) or denaturing (Group 2) conditions was obtained (P > .5).

Bottom Line: Our previous results showed that a soluble fraction from adult stages of the nematode (p26/23) induced partial protection against challenge.Recombinant DNA technology was applied to obtain a synthetic protein (rHcp26/23).Immunization did not induce any significant protection after challenge with 16000 infective larvae of H. contortus, and comparable values for parasite faecal egg output, packed cell volume, and abomasal parasite burdens were found in vaccinated and control animals.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.

ABSTRACT
Haemonchosis, caused by the abomasal nematode Haemonchus contortus, is a common parasitic disease of sheep. Our previous results showed that a soluble fraction from adult stages of the nematode (p26/23) induced partial protection against challenge. Recombinant DNA technology was applied to obtain a synthetic protein (rHcp26/23). Immunological assays (ELISA, Western blotting, and immunolocalization), using sera from lambs immunized with p26/23, confirmed the identity of the recombinant protein and demonstrated that the synthetic protein is equivalent to the purified protein employed in the previous immunoprophylaxis studies. Vaccination of lambs with 300 mug of rHcp26/23 and Freund's adjuvant elicited a notable specific antibody response. Immunization did not induce any significant protection after challenge with 16000 infective larvae of H. contortus, and comparable values for parasite faecal egg output, packed cell volume, and abomasal parasite burdens were found in vaccinated and control animals.

No MeSH data available.


Related in: MedlinePlus