Authors: Cacciabue, Marco Polo Domingo; García Núñez, María Soledad; Delgado, Fernando Oscar; Currá, Anabella Paola; Marrero, Ruben; Molinari, Maria Paula; Rieder, Elizabeth; Carrillo, Elisa Cristina; Gismondi, Maria Ines
Publication Date: 2017.
Language: English.
Abstract:
Adult C57BL/6J mice have been used to study Foot-and-mouth disease virus (FMDV) biology. In this work, two variants of an FMDV A/Arg/01 strain exhibiting differential pathogenicity in adult mice were identified and characterized: a non-lethal virus (A01NL) caused mild signs of disease, whereas a lethal virus (A01L) caused death within 24–48 h independently of the dose used. Both viruses caused a systemic infection with pathological changes in the exocrine pancreas. Virus A01L reached higher viral loads in plasma and organs of inoculated mice as well as increased replication in an ovine kidney cell line. Complete consensus sequences revealed 6 non-synonymous changes between A01L and A10NL genomes that might be linked to replication differences, as suggested by in silico prediction studies. Our results highlight the biological significance of discrete genomic variations and reinforce the usefulness of this animal model to study viral determinants of lethality.
Author affiliation: Cacciabue, Marco Polo Domingo. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: García Núñez, María Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Delgado, Fernando Oscar. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Author affiliation: Currá, Anabella Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Marrero, Ruben. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Molinari, Maria Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Rieder, Elizabeth. United States Department of Agriculture; Argentina
Author affiliation: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Gismondi, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Cacciabue, Marco Polo; Garcia Nuñez, Maria Soledad; Delgado, Fernando Oscar; Curra, Anabella Paola; Marrero Diaz De Villegas, Rubén; Molinari, Maria Paula; Rieder, Elizabeth; Carrillo, Elisa Cristina; Gismondi, Maria Ines
Publication Date: 2017.
Language: English.
Abstract:
Adult C57BL/6J mice have been used to study Foot-and-mouth disease virus (FMDV) biology. In this work, two variants of an FMDV A/Arg/01 strain exhibiting differential pathogenicity in adult mice were identified and characterized: a non-lethal virus (A01NL) caused mild signs of disease, whereas a lethal virus (A01L) caused death within 24–48 h independently of the dose used. Both viruses caused a systemic infection with pathological changes in the exocrine pancreas. Virus A01L reached higher viral loads in plasma and organs of inoculated mice as well as increased replication in an ovine kidney cell line. Complete consensus sequences revealed 6 nonsynonymous changes between A01L and A10NL genomes that might be linked to replication differences, as suggested by in silico prediction studies. Our results highlight the biological significance of discrete genomic variations and reinforce the usefulness of this animal model to study viral determinants of lethality.
Inst. de Biotecnología
Author affiliation: Cacciabue, Marco Polo. INTA. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Garcia Nuñez, Maria Soledad. INTA. Instituto de Biotecnología; Argentina
Author affiliation: Delgado, Fernando Oscar. INTA. Instituto de Patobiología; Argentina
Author affiliation: Curra, Anabella Paola. INTA. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Marrero Diaz De Villegas, Rubén. INTA. Instituto de Biotecnología; Argentina
Author affiliation: Molinari, Maria Paula. INTA. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Carrillo, Elisa Cristina. INTA. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Gismondi, Maria Ines. INTA. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Keywords: Ratón; Fiebre Aftosa; Patogénesis; Mice; Foot and Mouth Disease; Pathogenesis.
Repository: INTA Digital (INTA). Instituto Nacional de Tecnología Agropecuaria
Authors: García Núñez, María Soledad; Gismondi, Maria Ines; König, Guido Alberto; Berinstein, Analía; Taboga, Oscar Alberto; Rieder, Elizabeth; Martínez Salas, Encarnación; Carrillo, Elisa Cristina
Publication Date: 2014.
Language: English.
Abstract:
A reverse genetics approach was used to identify viral genetic determinants of the differential virulence displayed by two field foot-and-mouth disease virus (FMDV) strains (A/Arg/00 and A/Arg/01) isolated in Argentina during the 2000-2001 epidemics. A molecular clone of A/Arg/01 strain and viral chimeras containing the S-fragment or the internal ribosome entry site (IRES) of A/Arg/00 in the A/Arg/01 backbone were constructed and characterized. The IRES appeared as a determining factor of the lower level of A/Arg/00 replication in cell culture. High-throughput RNA probing revealed structural differences between both IRESs. Translation experiments using either synthetic viral RNAs ( in vitro) or bicistronic plasmids ( in vivo) showed that these IRESs' activities differ when the viral 3' untranslated region (UTR) is present, suggesting that their function is differentially modulated by this region. This work provides experimental evidence supporting the role of the IRES-3'UTR modulation in determining the level of FMDV replication in field strains.
Author affiliation: García Núñez, María Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Gismondi, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: König, Guido Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Berinstein, Analía. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Author affiliation: Rieder, Elizabeth. Plum Island Animal Disease Center; Estados Unidos. United States Department of Agriculture. Agricultural Research Service; Argentina
Author affiliation: Martínez Salas, Encarnación. Universidad Autónoma de Madrid. Centro de Biología Molecular; España. Consejo Superior de Investigaciones Científicas; España
Author affiliation: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Kenney, Mary; Waters, Ryan A.; Rieder, Elizabeth; Pega, Juan Franco; Pérez Filgueira, Daniel Mariano; Golde, William T.
Publication Date: 2017.
Language: English.
Abstract:
Analysis of the immune response to infection of livestock by foot-and-mouth disease virus (FMDV) is most often reported as the serum antibody response to the virus. While measurement of neutralizing antibody has been sensitive and specific, measurements of the quality of the antibody response are less robust. Determining the immunoglobulin (Ig) isotype of the serum antibody response provides a deeper understanding of the biology of the response and more sensitive methods for these assays will facilitate analyses of B cell mediated immunity. We tested the hypothesis that using the virus as the molecular probe could be achieved by adding tags to the surface of the FMDV capsid, and that would enhance sensitivity in assays for anti-FMDV antibody responses. The use of a FLAG-tagged virus in these assays failed to yield improvement whereas chemically biotinylating the virus capsid resulted in significant enhancement of the signal. Here we describe methods using biotinylated virus for measuring anti-viral antibody in serum and antibody secreting cells (ASCs) in blood that are sensitive and specific. Finally, we describe using the biotinylated virus in flow cytometry where such assays should greatly enhance the analysis of anti-virus antibody producing B cells, allowing the investigator to focus on only the FMDV specific B cells when analyzing the development of the B cell response to either infection or vaccination.
Author affiliation: Kenney, Mary. Agricultural Research Service; Estados Unidos
Author affiliation: Waters, Ryan A.. Agricultural Research Service; Estados Unidos
Author affiliation: Rieder, Elizabeth. Agricultural Research Service; Estados Unidos
Author affiliation: Pega, Juan Franco. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
Author affiliation: Pérez Filgueira, Daniel Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
Author affiliation: Golde, William T.. Agricultural Research Service; Estados Unidos
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas
Authors: Kenney, Mary; Waters, Ryan A.; Rieder, Elizabeth; Pega, Juan Franco; Perez Filgueira, Daniel Mariano; Golde, William T.
Publication Date: 2017.
Language: English.
Abstract:
Analysis of the immune response to infection of livestock by foot-and-mouth disease virus (FMDV) is most often reported as the serum antibody response to the virus. While measurement of neutralizing antibody has been sensitive and specific, measurements of the quality of the antibody response are less robust. Determining the immunoglobulin (Ig) isotype of the serum antibody response provides a deeper understanding of the biology of the response and more sensitive methods for these assays will facilitate analyses of B cell mediated immunity. We tested the hypothesis that using the virus as the molecular probe could be achieved by adding tags to the surface of the FMDV capsid, and that would enhance sensitivity in assays for anti-FMDV antibody responses. The use of a FLAG-tagged virus in these assays failed to yield improvement whereas chemically biotinylating the virus capsid resulted in significant enhancement of the signal. Here we describe methods using biotinylated virus for measuring anti-viral antibody in serum and antibody secreting cells (ASCs) in blood that are sensitive and specific. Finally, we describe using the biotinylated virus in flow cytometry where such assays should greatly enhance the analysis of anti-virus antibody producing B cells, allowing the investigator to focus on only the FMDV specific B cells when analyzing the development of the B cell response to either infection or vaccination.
Instituto de Virología
Author affiliation: Kenney, Mary. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos
Author affiliation: Waters, Ryan A. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos
Author affiliation: Rieder, Elizabeth. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos
Author affiliation: Pega, Juan Franco. INTA. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Perez Filgueira, Daniel Mariano. INTA. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Author affiliation: Golde, William T. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos
Repository: INTA Digital (INTA). Instituto Nacional de Tecnología Agropecuaria