Authors: Hecker, Yanina Paola; Cóceres, Verónica Mabel; Wilkowsky, Silvina Elizabeth; Jaramillo Ortiz, José Manuel; Morrell, Eleonora Lidia; Verna, Andrea Elizabeth; Ganuza, Agustina; Cano, Dora B.; Lischinsky, Lilian; Ángel, Sergio Oscar; Zamorano, Patricia Ines; Odeón, Anselmo Carlos; Leunda, Maria Rosa; Campero, Carlos Manuel; Morein, Bror; Moore, Dadin Prando
Publication Date: 2014.
Language: English.
Abstract:
The aim of the present study was to evaluate the immunogenicity and protective efficacy of rNcSAG1, rNcHSP20, rNcGRA7 recombinant proteins formulated with immune stimulating complexes (ISCOMs) in pregnant heifers against vertical transmission of N. caninum. Twelve pregnant heifers were divided into 3 groups of 4 heifers each, receiving different formulations before mating. Immunogens were administered twice subcutaneously: group A animals were inoculated with three recombinant proteins (rNcSAG1, rNcHSP20, rNcGRA7) formulated with ISCOMs; group B animals received ISCOM-MATRIX (without antigen) and group C received sterile phosphate-buffered saline (PBS) only. The recombinant proteins were expressed in Escherichia coli and purified by nickel resin. All groups were intravenously challenged with the NC-1 strain of Neospora caninum at day 70 of gestation and dams slaughtered at week 17 of the experiment. Heifers from group A developed specific antibodies against rNcSAG1, rNcHSP20 and rNcGRA7 prior to the challenge. Following immunization, an statistically significant increase of antibodies against rNcSAG1 and rNcHSP20 in all animals of group A was detected compared to animals in groups B and C at weeks 5, 13 and 16 (P <0.001). Levels of antibodies against rNcGRA7 were statistical higher in group A animals when compared with groups B and C at weeks 5 and 16 (P > 0.001). There were no differences in IFN-γ production among the experimental groups at any time point (P > 0.05). Transplacental transmission was determined in all fetuses of groups A, B and C by Western Blot, Immunohistochemistry and nested PCR. This work showed that rNcSAG1, rNcHSP20 and rNcGRA7 proteins while immunogenic in cattle failed to prevent the fetal infection in pregnant cattle challenged at Day 70 of gestation.
Author affiliation: Hecker, Yanina Paola. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Cóceres, Verónica Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; Argentina
Author affiliation: Wilkowsky, Silvina Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Jaramillo Ortiz, José Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Morrell, Eleonora Lidia. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Verna, Andrea Elizabeth. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Ganuza, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Author affiliation: Cano, Dora B.. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Lischinsky, Lilian. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Ángel, Sergio Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Author affiliation: Zamorano, Patricia Ines. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentina
Author affiliation: Morein, Bror. Uppsala University. Department of Medical Sciences. Infectious Diseases; Suecia
Author affiliation: Moore, Dadin Prando. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas