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Induction of immunity in sheep to Fasciola hepatica with mimotopes of cathepsin L selected from a phage display library

Published online by Cambridge University Press:  24 September 2008

A. VILLA-MANCERA ,
H. QUIROZ-ROMERO ,
D. CORREA ,
F. IBARRA ,
M. REYES-PÉREZ ,
H. REYES-VIVAS ,
G. LÓPEZ-VELÁZQUEZ ,
K. GAZARIAN ,
T. GAZARIAN  and
R. A. ALONSO
A. VILLA-MANCERA
Affiliation:
Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510
H. QUIROZ-ROMERO*
Affiliation:
Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510
D. CORREA
Affiliation:
Laboratorio de Inmunología Experimental, Instituto Nacional de Pediatría, México, D.F. 04530
F. IBARRA
Affiliation:
Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510
M. REYES-PÉREZ
Affiliation:
Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510
H. REYES-VIVAS
Affiliation:
Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, México, D.F. 04530
G. LÓPEZ-VELÁZQUEZ
Affiliation:
Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, México, D.F. 04530
K. GAZARIAN
Affiliation:
Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, UNAM, México, D.F. 04510
T. GAZARIAN
Affiliation:
Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, UNAM, México, D.F. 04510
R. A. ALONSO
Affiliation:
Departamento de Genética y Bioestadística, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510
*
*Corresponding author: Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, UNAM, México, D.F. 04510. Tel: +52 56225898. Fax: +52 56225898. E-mail: [email protected]
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Summary

An M13 phage random 12-mers peptide library was used to screen cathepsin L mimotopes of Fasciola hepatica and to evaluate their immunogenicity in sheep. Seven clones showed positive reactivity to a rabbit anti-cathepsin L1/L2 antiserum in ELISA, and their amino acid sequences deduced by DNA sequencing were tentatively mapped on the protein. Twenty sheep were randomly allocated into 4 groups of 5 animals each, for immunization with 1×1014 phage particles of clones 1, 20, a mixture of 7 clones and PBS, without adjuvant at the beginning, and 4 weeks later. All groups were challenged with 300 metacercariae at week 6 and slaughtered 16 weeks later. The mean worm burdens after challenge were reduced by 47·61% and 33·91% in sheep vaccinated with clones 1 and 20, respectively; no effect was observed in animals inoculated with the clone mixture. Also, a significant reduction in worm size and burden was observed for those sheep immunized with clone 1. Animals receiving clone 20, showed a significant reduction in egg output. Immunization induced a reduction of egg viability ranging from 58·92 to 82·11%. Furthermore, vaccinated animals produced clone-specific antibodies which were boosted after challenge with metacercariae of F. hepatica.

Keywords

Fasciola hepaticacathepsin Lphage displaymimotopesvaccine
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 12 , October 2008 , pp. 1437 - 1445
Copyright
Copyright © 2008 Cambridge University Press

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