المؤلفون: Currá, Anabella, Cacciabue, Marco, José Gravisaco, María, Asurmendi, Sebastián, Taboga, Oscar, Gismondi, María I.

المصدر: PeerJ; Jun2021, p1-18, 18p

مصطلحات موضوعية: FOOT & mouth disease, RNA replicase, VIRUS diseases, NON-coding RNA, MICRORNA, RNA polymerases

مستخلص: RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causative agent of a devastating animal disease with high economic impact. The aim of this work was to assess the antiviral activity of different small RNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D). Three target sequences were predicted within 3D considering RNA accessibility as a major criterion. The silencing efficacy of shorthairpin RNAs (shRNAs) and artificial microRNAs (amiRNAs) targeting the selected sequences was confirmed in fluorescent reporter assays. Furthermore, BHK-21 cells transiently expressing shRNAs or amiRNAs proved 70 to >95% inhibition of FMDV growth. Interestingly, dual expression of amiRNAs did not improve FMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs were established and characterized in terms of antiviral activity against FMDV. As expected, viral replication in these cell lines was delayed. These results show that the target RNA-accessibilityguided approach for RNAi design rendered efficient amiRNAs that constrain FMDV replication. The application of amiRNAs to complement FMDV vaccination in specific epidemiological scenarios shall be explored further. [ABSTRACT FROM AUTHOR]

: Copyright of PeerJ is the property of PeerJ Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Diez, Michay, Trotta, Myrian, Alfonso, Victoria, Taboga, Oscar, López, María Gabriela

المصدر: 3 Biotech; 10/20/2018, Vol. 8 Issue 11, p1-1, 1p

مصطلحات موضوعية: ENZYME-linked immunosorbent assay, FOOT & mouth disease, POLYPEPTIDES, COMMUNICABLE diseases, ANTIGENS

مستخلص: Here, we developed a diagnostic ELISA for foot-and-mouth disease using recombinant occlusion bodies (rOBs) of baculovirus. We fused Δ3AB1-3, a polypeptide derived from non-structural proteins of foot-and-mouth disease virus, to polyhedrin (POLH), the major constituent of OBs, under polh promoter. To further assess the most convenient strategy to improve yields, we designed two recombinant baculoviruses, vPOLH and vPOLHE44G. These carried the sequence of the fusion protein POLH-Δ3AB1-3 with an additional copy in cis of polh or polhE44G, respectively, under p10 promoter. Our results show that both viruses expressed POLH-Δ3AB1-3, which was detected by western blot in purified rOBs with anti-POLH and anti-3AB1 antibodies. We also found that vPOLHE44G produced larger polyhedra and a significant increase of antigen yield (p < 0.01). Furthermore, the chimeric protein POLH-Δ3AB1-3 was recognized by sera from experimentally infected animals, showing that translational fusion to POLH does not alter the antigenicity of Δ3AB1-3. Finally, the rOBs were successfully used in an ELISA test to differentiate infected from vaccinated animals. Taken together, these results demonstrate the great potential of rOBs to develop diagnostic schemes adaptable to animal infectious diseases. [ABSTRACT FROM AUTHOR]

: Copyright of 3 Biotech is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: García-Nuñez, Soledad¹ mgnunez@cnia.inta.gov.ar, Gismondi, María Inés¹ mgismondi@cnia.inta.gov.ar, König, Guido¹ gkonig@cnia.inta.gov.ar, Berinstein, Analía¹ aberinstein@cnia.inta.gov.ar, Taboga, Oscar¹ otaboga@cnia.inta.gov.ar, Rieder, Elizabeth² elizabeth.rieder@ars.usda.gov, Martínez-Salas, Encarnación³ emartinez@cbm.uam.es, Carrillo, Elisa¹ ecarrillo@cnia.inta.gov.ar

المصدر: Virology. Jan2014, Vol. 448, p303-313. 11p.

مصطلحات موضوعية: *RIBOSOMES, *NON-coding RNA, *MICROBIAL virulence, *FOOT & mouth disease, *VIRAL replication, *CELL culture

مستخلص: 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. [Copyright &y& Elsevier]

المؤلفون: López, María Gabriela¹, Peralta, Andrea^1,2, Berinstein, Analía^1,2, Fondevila, Norberto³, Carrillo, Elisa^1,2, Taboga, Oscar¹ otaboga@cnia.inta.gov.ar

المصدر: Journal of Virological Methods. Mar2005, Vol. 124 Issue 1/2, p221-224. 4p.

مصطلحات موضوعية: *FOOT & mouth disease, *VIRUS diseases, *GENETICS, *ENZYME-linked immunosorbent assay

مستخلص: Abstract: For its potential usefulness in diagnosis, the non-structural protein 3AB1 from foot-and-mouth disease virus was expressed as a soluble protein by using Autographa californica nuclear polyhedrosis virus as a vector. The 3AB1 coding sequence was introduced into AcNPV genome via pBAcPAK3AB1 transfer vector to originate Ac3AB1 recombinant baculovirus of phenotype occ−. Rachiplusia nu larvae were injected with supernatants of Sf9 cells infected with Ac3AB1 and 5 days post-infection total protein extracts were obtained. An intense band of approximately 21.5kDa was observed when total larvae extracts were SDS-PAGE resolved and the recombinant protein detected by an FMDV-infected guinea pig serum. ELISA tests and Western blot experiments were carried out using sera both from FMDV-infected cattle and from vaccinated animals. The recombinant protein was only recognized by sera from infected animals, suggesting that this method of production in insect larvae could be applied to an efficient mass production of proteins of diagnostic interest. [Copyright &y& Elsevier]

المؤلفون: Molina, Guido Nicolás¹ (AUTHOR), Cacciabue, Marco¹ (AUTHOR), Gismondi, María Inés¹ (AUTHOR), Taboga, Oscar¹ (AUTHOR), Molinari, Paula¹ (AUTHOR) molinari.maria@inta.gob.ar

المصدر: Antiviral Research. Aug2020, Vol. 180, pN.PAG-N.PAG. 1p.

مصطلحات موضوعية: *TYPE I interferons, *FOOT & mouth disease, *VIRUS diseases, *MOUSE diseases, *KILLER cells, *INTERFERON receptors

مستخلص: Foot-and-mouth disease is a viral illness that affects cloven-hoofed animals causing serious economic losses. Inactivated vaccines against its causative agent, foot-and-mouth disease virus (FMDV), require approximately seven days to induce protection. Therefore, antiviral strategies are needed to provide earlier protection and to stop the spread of this highly contagious virus during outbreak situations. In this way, our group has previously demonstrated that the baculovirus (BV) Autographa californica multiple nucleopolyhedrovirus (AcMNPV), an insect virus with immunostimulant effects, induces a nonspecific antiviral status that protects C57BL/6 mice against a lethal challenge with FMDV A/Arg/01 at 3 hours or 3 days post inoculation. In this work, we studied the immunological mechanisms involved in this protection. Firstly, we compared the protection elicited by AcMNPV in wild type mice and in knock-out mice lacking the subunit IFNAR1 of the receptor for type I interferons (IFNs). Our results showed that type I IFNs are key to prevent the death of the animals after the FMDV challenge. On the other hand, we evaluated the role of NK and NKT cells by depleting these cell subsets with anti-NK1.1 monoclonal antibody. These cells proved to be necessary for the induction of IFN-γ by AcMNPV and to prevent the onset of a severe disease after the FMDV challenge. We propose BV as a novel tool for the development of antiviral strategies because of the high levels of IFNs induced and the NK/NKT cells-mediated immune response elicited. Image 1 • Baculovirus promotes early and nonspecific protection against FMDV in mice. • This antiviral status is mainly mediated by the type I IFN system. • NK cells are necessary for the induction of type II IFNs. • NK cells are needed to prevent the onset of the disease. [ABSTRACT FROM AUTHOR]