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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

Immunolocalization of p26/23 in histological sections of adult H. contortus. Fluorescence was detected with antinative p26/23 hyperimmune rabbit serum and an anti-rHcp26/23 hyperimmune rabbit serum. Antirabbit Cy3-conjugated was the secondary antibody. Sections of adult H. contortus showed p26/23 recognition in the hypodermic chords of the nematode with anti-p26/23 (a) and anti-rHcp26/23 (b). Section incubated with normal rabbit serum (c).
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fig3: Immunolocalization of p26/23 in histological sections of adult H. contortus. Fluorescence was detected with antinative p26/23 hyperimmune rabbit serum and an anti-rHcp26/23 hyperimmune rabbit serum. Antirabbit Cy3-conjugated was the secondary antibody. Sections of adult H. contortus showed p26/23 recognition in the hypodermic chords of the nematode with anti-p26/23 (a) and anti-rHcp26/23 (b). Section incubated with normal rabbit serum (c).

Mentions: WB analysis of rHcp26/23, and soluble extracts from both infective larvae (LSE) and adult stages (ASE) of H. contortus with sera from rabbit antinative p26/23, rabbit anti-rHcp26/23, and lamb anti-p26/23 fraction, showed comparable reactivity patterns irrespective of the antigen source (LSE, ASE). A clear band, ca. 24–26 kDa, was developed in both stages of the helminth, using the serum against the native p26/23 (Figures 2(a) and 2(b), lane 1) or the anti-rHcp26/23 (Figures 2(a) and 2(b), lane 2). It was also observed that the recombinant protein was recognized by the sera from vaccinated lambs (p26/23) (Figure 2(c), lane 3). Some reactivity was found at ca. 46 kDa, particularly when ASE (Figure 2(a)) and rHcp26/23 (Figure 2(c)) were probed with immune sera. Antigenic similarity between p26/23 and rHcp26/23 was confirmed by immunolocalization studies. Both hyperimmune sera (anti-p26/23 and anti-rHcp26/23), but not the serum from nonimmunized rabbit, specifically reacted against the hypodermic chords of adult H. contortus sections (Figure 3).


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)

Immunolocalization of p26/23 in histological sections of adult H. contortus. Fluorescence was detected with antinative p26/23 hyperimmune rabbit serum and an anti-rHcp26/23 hyperimmune rabbit serum. Antirabbit Cy3-conjugated was the secondary antibody. Sections of adult H. contortus showed p26/23 recognition in the hypodermic chords of the nematode with anti-p26/23 (a) and anti-rHcp26/23 (b). Section incubated with normal rabbit serum (c).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: Immunolocalization of p26/23 in histological sections of adult H. contortus. Fluorescence was detected with antinative p26/23 hyperimmune rabbit serum and an anti-rHcp26/23 hyperimmune rabbit serum. Antirabbit Cy3-conjugated was the secondary antibody. Sections of adult H. contortus showed p26/23 recognition in the hypodermic chords of the nematode with anti-p26/23 (a) and anti-rHcp26/23 (b). Section incubated with normal rabbit serum (c).
Mentions: WB analysis of rHcp26/23, and soluble extracts from both infective larvae (LSE) and adult stages (ASE) of H. contortus with sera from rabbit antinative p26/23, rabbit anti-rHcp26/23, and lamb anti-p26/23 fraction, showed comparable reactivity patterns irrespective of the antigen source (LSE, ASE). A clear band, ca. 24–26 kDa, was developed in both stages of the helminth, using the serum against the native p26/23 (Figures 2(a) and 2(b), lane 1) or the anti-rHcp26/23 (Figures 2(a) and 2(b), lane 2). It was also observed that the recombinant protein was recognized by the sera from vaccinated lambs (p26/23) (Figure 2(c), lane 3). Some reactivity was found at ca. 46 kDa, particularly when ASE (Figure 2(a)) and rHcp26/23 (Figure 2(c)) were probed with immune sera. Antigenic similarity between p26/23 and rHcp26/23 was confirmed by immunolocalization studies. Both hyperimmune sera (anti-p26/23 and anti-rHcp26/23), but not the serum from nonimmunized rabbit, specifically reacted against the hypodermic chords of adult H. contortus sections (Figure 3).

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