Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle

Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle

In order to evaluate the interferon gamma (IFN-g) induction ability of different proteic preparations and that of Brucella abortus smooth and rough lipopolysaccharides in whole blood cultures from RB51-immunized cattle, both vaccinated and revaccinated cattle were selected from a herd with high brucellosis prevalence. The assay was validated using a group of serologically-negative cattle from a control herd. Whole blood cultures were stimulated with the different antigens and the mitogen concanavalin A, for 24 h at 37 ºC. Plasma supernatants were collected to evaluate the IFN-g released, using a commercially-available sandwich ELISA kit. Results expressed as stimulation indexes showed a higher cytokine-inducing ability for the sonicated extract followed by cytosol proteins and inner membrane protein, for the different groups. Nevertheless, even though boosted, serologically-positive animal showed a significant response lower than that observed in the serologically-negative groups, probably due to a shift in the prevalence of B. abortus antigen-responsive cell subpopulations, indirectly revealed by the presence of high antibody levels. Therefore, the usefulness of estimating IFN-g release after the in vitro stimulation of whole blood cultures with a sonicated B. abortus extract for the evaluation and follow up of cellular immunity in vaccinated cows was demonstrated.

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Bibliographic Details
Main Authors: Leal Hernández, Marisela, Jaramillo Meza, Laura, Hernández Andrade, Laura, Pérez González, Rafael, Díaz Aparicio, Efrén, Suárez Güemes, Francisco
Format: Digital revista
Language:spa
Published: Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias 2012
Online Access:https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1779
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id rev-remexcp-article1779
record_format ojs
institution INIFAP
collection OJS
country México
countrycode MX
component Revista
access En linea
databasecode rev-remexcp
tag revista
region America del Norte
libraryname Biblioteca INIFAP
language spa
format Digital
author Leal Hernández, Marisela
Jaramillo Meza, Laura
Hernández Andrade, Laura
Pérez González, Rafael
Díaz Aparicio, Efrén
Suárez Güemes, Francisco
spellingShingle Leal Hernández, Marisela
Jaramillo Meza, Laura
Hernández Andrade, Laura
Pérez González, Rafael
Díaz Aparicio, Efrén
Suárez Güemes, Francisco
Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
author_facet Leal Hernández, Marisela
Jaramillo Meza, Laura
Hernández Andrade, Laura
Pérez González, Rafael
Díaz Aparicio, Efrén
Suárez Güemes, Francisco
author_sort Leal Hernández, Marisela
title Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
title_short Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
title_full Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
title_fullStr Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
title_full_unstemmed Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle
title_sort production of interferon gamma in whole blood cultures in response to brucella abortus antigens in rb51-vaccinated cattle
description In order to evaluate the interferon gamma (IFN-g) induction ability of different proteic preparations and that of Brucella abortus smooth and rough lipopolysaccharides in whole blood cultures from RB51-immunized cattle, both vaccinated and revaccinated cattle were selected from a herd with high brucellosis prevalence. The assay was validated using a group of serologically-negative cattle from a control herd. Whole blood cultures were stimulated with the different antigens and the mitogen concanavalin A, for 24 h at 37 ºC. Plasma supernatants were collected to evaluate the IFN-g released, using a commercially-available sandwich ELISA kit. Results expressed as stimulation indexes showed a higher cytokine-inducing ability for the sonicated extract followed by cytosol proteins and inner membrane protein, for the different groups. Nevertheless, even though boosted, serologically-positive animal showed a significant response lower than that observed in the serologically-negative groups, probably due to a shift in the prevalence of B. abortus antigen-responsive cell subpopulations, indirectly revealed by the presence of high antibody levels. Therefore, the usefulness of estimating IFN-g release after the in vitro stimulation of whole blood cultures with a sonicated B. abortus extract for the evaluation and follow up of cellular immunity in vaccinated cows was demonstrated.
publisher Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias
publishDate 2012
url https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1779
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spelling rev-remexcp-article17792020-06-18T19:38:29Z Production of interferon gamma in whole blood cultures in response to Brucella abortus antigens in RB51-vaccinated cattle Producción de interferón gamma en cultivos de sangre completa en respuesta a antígenos de Brucella abortus en bovinos vacunados con RB51 Leal Hernández, Marisela Jaramillo Meza, Laura Hernández Andrade, Laura Pérez González, Rafael Díaz Aparicio, Efrén Suárez Güemes, Francisco Cattle, Brucella abortus, Brucellosis, Cellular Immunity, Interferon gamma Bovinos, Brucella abortus, Brucelosis, Inmunidad celular, Interferón gamma, RB51 In order to evaluate the interferon gamma (IFN-g) induction ability of different proteic preparations and that of Brucella abortus smooth and rough lipopolysaccharides in whole blood cultures from RB51-immunized cattle, both vaccinated and revaccinated cattle were selected from a herd with high brucellosis prevalence. The assay was validated using a group of serologically-negative cattle from a control herd. Whole blood cultures were stimulated with the different antigens and the mitogen concanavalin A, for 24 h at 37 ºC. Plasma supernatants were collected to evaluate the IFN-g released, using a commercially-available sandwich ELISA kit. Results expressed as stimulation indexes showed a higher cytokine-inducing ability for the sonicated extract followed by cytosol proteins and inner membrane protein, for the different groups. Nevertheless, even though boosted, serologically-positive animal showed a significant response lower than that observed in the serologically-negative groups, probably due to a shift in the prevalence of B. abortus antigen-responsive cell subpopulations, indirectly revealed by the presence of high antibody levels. Therefore, the usefulness of estimating IFN-g release after the in vitro stimulation of whole blood cultures with a sonicated B. abortus extract for the evaluation and follow up of cellular immunity in vaccinated cows was demonstrated. Con el propósito de evaluar la capacidad inductora de la producción de interferón gama de diferentes preparaciones proteínicas, y de los lipopolisacáridos liso y rugoso de Brucella abortus en cultivos de sangre completa de bovinos inmunizados con RB51, se seleccionaron animales vacunados y revacunados de un hato con elevada prevalencia de la enfermedad, igualmente, un grupo de animales seronegativos de un hato control fue considerado para valorar la utilidad del ensayo. Cultivos de sangre completa de los animales se estimularon con los antígenos, y con el mitogeno concanavalina A, por un periodo de 24 h a 37 ºC, posteriormente se colectaron los plasmas para evaluar el IFN-g liberado, empleando un sistema comercial de ELISA-sandwich. Los resultados expresados en índices de estimulación indican mayor capacidad inductora de la citocina para el extracto sonicado, seguida de las proteínas de citosol y las de membrana interna, para los diferentes grupos, no obstante, en el grupo de animales seropositivos revacunados a pesar de observar una respuesta significativa, ésta fue menor a la que se presentó en los grupos seronegativos, debida probablemente a un cambio en el dominio de las subpoblaciones celulares responsivas a los antígenos de Brucella, revelado indirectamente por la presencia de los niveles altos de anticuerpos mostrados. Así, se demuestra que la estimación del INF-g liberado luego de la estimulación in vitro de cultivos de sangre completa empleando un extracto sonicado de Brucella abortus, es de utilidad para la evaluación y seguimiento de la inmunidad celular en animales vacunados. Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias 2012-01-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1779 Revista Mexicana de Ciencias Pecuarias; Vol. 45, Núm. 2 (2007); 147 a 159 Revista Mexicana de Ciencias Pecuarias; Vol. 45, Núm. 2 (2007); 147 a 159 2448-6698 2007-1124 spa https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1779/1773 http://creativecommons.org/licenses/by-nc-sa/4.0

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