A DNA vaccine encoding foot-and-mouth disease virus B and T-cell epitopes targeted to class II swine leukocyte antigens protects pigs against viral challenge

Development of efficient and safer vaccines against foot-and-mouth disease virus (FMDV) is a must. Previous results obtained in our laboratory have demonstrated that DNA vaccines encoding B and T cell epitopes from type C FMDV, efficiently controlled virus replication in mice, while they did not protect against FMDV challenge in pigs, one of the FMDV natural hosts. The main finding of this work is the ability to improve the protection afforded in swine using a new DNA-vaccine prototype (pCMV-APCH1BTT), encoding FMDV B and T-cell epitopes fused to the single-chain variable fragment of the 1F12 mouse monoclonal antibody that recognizes Class-II Swine Leukocyte antigens. Half of the DNA-immunized pigs were fully protected upon viral challenge, while the remaining animals were partially protected, showing a delayed, shorter and milder disease than control pigs. Full protection in a given vaccinated-pig correlated with the induction of specific IFNγ-secreting T-cells, detectable prior to FMDV-challenge, together with a rapid development of neutralizing antibodies after viral challenge, pointing towards the relevance that both arms of the immune response can play in protection. Our results open new avenues for developing future FMDV subunit vaccines.Fil: Borrego, Belén. No especifíca;Fil: Argilaguet, Jordi M.. Universitat Autònoma de Barcelona; EspañaFil: Pérez Martín, Eva. Universitat Autònoma de Barcelona; EspañaFil: Dominguez, Javier. No especifíca;Fil: Pérez Filgueira, Daniel Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Escribano, José M.. No especifíca;Fil: Sobrino, Francisco. No especifíca;Fil: Rodriguez, Fernando. Universitat Autònoma de Barcelona; Españ

Editorial : Foot-and-mouth disease epidemiology, vaccines and vaccination : moving forward

Vaccination has played a major role in foot-and-mouth disease (FMD) control. There are different approaches to the design and implementation of vaccination campaigns, and epidemiological information is paramount in influencing the vaccine and vaccination strategy that best suit each geographic location. FMD-endemic regions typically organize vaccination campaigns as a routine preventive control policy or to mitigate the impact of the disease. The majority of currently used vaccines are formulated with chemically inactivated whole-viral particles and suitable adjuvants such as single and double oil emulsions. The most recent strains circulating in a particular region are typically selected as antigens based on the results of vaccine-matching data and in vitro experiments, however, predictions based on vaccine-matching approaches are usually uncertain without a live virus challenge in natural hosts combined with reliable field data. Vaccine selection and successful vaccination campaigns rely on a deep knowledge of the epidemiology of the region where these vaccines will be used, as well as access to the appropriate diagnostic tools to underpin these campaigns.Instituto de VirologíaFil: Capozzo, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Capozzo, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vosloo, Wilna. Commonwealth Scientific and Industrial Research Organisation (CSIRO) Health and Biosecurity. Transboundary Disease Mitigation. Australian Centre for Disease Preparedness; AustraliaFil: de los Santos, Teresa. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados UnidosFil: Pérez, Andrés M. University of Minnesota. Department of Veterinary Population Medicine; Estados UnidosFil: Perez Filgueira, Daniel Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Perez Filgueira, Daniel Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Enhancing dna immunization by targeting asfv antigens to sla-ii bearing cells

One of the main criticisms to DNA vaccines is the poor immunogenicity that they confer on occasions, at least in large animals. Confirming this theory, immunization with plasmid DNA encoding two African swine fever virus genes in frame (pCMV-PQ), failed in inducing detectable immune responses in pigs, while it was successful in mice. Aiming to improve the immune responses induced in swine, a new plasmid was constructed, encoding the viral genes fused in frame with a single chain variable fragment of an antibody specific for a swine leukocyte antigen II (pCMV-APCH1PQ). Our results clearly demonstrate that targeting antigens to antigen professional cells exponentially enhanced the immune response induced in pigs, albeit that the DNA vaccine was not able to confer protection against lethal viral challenge. Indeed, a viremia exacerbation was observed in each of the pigs that received the pCMV-APCH1PQ plasmid, this correlating with the presence of non-neutralizing antibodies and antigen-specific SLA II-restricted T-cells. The implications of our discoveries for the development of future vaccines against African swine fever virus and other swine pathogens are discussed.Fil: Argilaguet, J. M.. No especifíca;Fil: Pérez Martín, E.. No especifíca;Fil: Gallardo, C.. No especifíca;Fil: Salguero, F. J.. No especifíca;Fil: Borrego, B.. No especifíca;Fil: Lacasta, A.. No especifíca;Fil: Accensi, F.. No especifíca;Fil: Díaz, I.. No especifíca;Fil: Nofrarías, M.. No especifíca;Fil: Pujols, J.. No especifíca;Fil: Blanco, E.. No especifíca;Fil: Pérez Filgueira, Daniel Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Escribano, José M.. No especifíca;Fil: Rodríguez, F.. No especifíca

Tasas de erosión, transporte y sedimentación en un medio estuárico: el Río Miñor (Ría de Vigo, Pontevedra)

The Miñor river is one o f the 35 minor currents entering the Ría de Vigo (Pontevedra) at Baiona Bay,; close to the SE margin o f the Ría. The present study was carried out over two periods: from May 97 to July 98 and from May 00 to May 01. In this paper we carry out a hydraulic and sedimentologic approach o f the Miñor river, making an estimation of the total fresh-water discharge and sediment input transported as suspended and dissolved loads. The final aim of this study is to calculate erosion rates at the catchment, sediment yield, transport and the nature of the sediments accumulated at the estuarine area. In view of the average discharge for both periods o f time (2.7 m3/s y 6.7 m3/s) and the catchment area (75.6 m2) is possible to estimate suspended and dissolved sediment yields. Mean values for suspended sediments are 25.76 t/km2a for 97/98 and 139.74 t/ km2a for 00/01 and those for dissolved loads of 109 t/km2a y 204 t/km2a, respectively. Finally, considering an average rock density of 2 .7 g/cm3 we can estimate mechanical denudation rates of 9.54 m3/km2a during 97/98 and 51.76 m3/km2a during 00/01. Chemical denudation rates are 40.46 m3/km2a for the first period and 75.56 m3/km2a for the second. Data obtained during 00/01 interval can be considered as maximum values, bearing in mind the very unusually high rainfall conditions

Silkworm expression system as a platform technology in life science

Many recombinant proteins have been successfully produced in silkworm larvae or pupae and used for academic and industrial purposes. Several recombinant proteins produced by silkworms have already been commercialized. However, construction of a recombinant baculovirus containing a gene of interest requires tedious and troublesome steps and takes a long time (3–6 months). The recent development of a bacmid, Escherichia coli and Bombyx mori shuttle vector, has eliminated the conventional tedious procedures required to identify and isolate recombinant viruses. Several technical improvements, including a cysteine protease or chitinase deletion bacmid and chaperone-assisted expression and coexpression, have led to significantly increased protein yields and reduced costs for large-scale production. Terminal N-acetyl glucosamine and galactose residues were found in the N-glycan structures produced by silkworms, which are different from those generated by insect cells. Genomic elucidation of silkworm has opened a new chapter in utilization of silkworm. Transgenic silkworm technology provides a stable production of recombinant protein. Baculovirus surface display expression is one of the low-cost approaches toward silkworm larvae-derived recombinant subunit vaccines. The expression of pharmaceutically relevant proteins, including cell/viral surface proteins, membrane proteins, and guanine nucleotide-binding protein (G protein) coupled receptors, using silkworm larvae or cocoons has become very attractive. Silkworm biotechnology is an innovative and easy approach to achieve high protein expression levels and is a very promising platform technology in the field of life science. Like the “Silkroad,” we expect that the “Bioroad” from Asia to Europe will be established by the silkworm expression system

Perfil de los pacientes que acuden al médico internista para valoración de osteoporosis: registro OSTEOMED

Producción CientíficaAntecedentes y objetivo: La osteoporosis se considera un trastorno generalizado del esqueleto en el que existe una alteración de la resistencia ósea que predispone a la persona a un mayor riesgo de fractura. Este estudio transversal pretende recoger y presentar las principales características clínicas de los pacientes que acuden a la consulta de los médicos internistas en Espa˜na. Conocer estas características podría facilitar la puesta en marcha de planes de actuación para mejorar la atención de estos pacientes de manera más eficaz y eficiente. Material y métodos: A través del análisis del registro OSTEOMED (Osteoporosis en Medicina Interna), este trabajo presenta las principales características clínicas de los pacientes con osteoporosis que acudieron a las consultas de Medicina Interna en 23 centros hospitalarios espa˜noles entre 2012 y 2017. Se han analizado los motivos de consulta, los valores densitométricos, la presencia de comorbilidades, el tratamiento prescrito y otros factores relacionados con el estilo de vida. Resultados: En total se evaluó a 2.024 pacientes con osteoporosis (89,87% mujeres, 10,13% hombres). La edad media de los pacientes fue de 64,1 ± 12,1 a˜nos (mujeres, 64,7 ± 11,5 a˜nos; hombres, 61,2 ± 14,2 a˜nos). No hubo diferencia entre sexos en la historia de caídas recientes (9,1-6,7%), mientras que sí se apreció en la ingesta diaria de calcio de lácteos (553,8 ± 332,6 mg en mujeres vs. 450,2 ± 303,3 mg en hombres; p < 0,001) y en causas secundarias de osteoporosis(13% de hombres vs. 6,5% de mujeres; p < 0,001). En la muestra se observaron un total de 404fracturas (20%), destacando el número de fracturas vertebrales confirmadas (17,2%, 35,6% enhombres vs. 15,2% de las mujeres; p < 0,001). Una gran parte de los pacientes no recibía eltratamiento indicado y presentaba bajos niveles de actividad física y exposición solar. Un por-centaje importante de pacientes presentó comorbilidades asociadas, siendo las más frecuentesla hipertensión (32%) y la dislipidemia (28%).Conclusiones: Estos resultados definen el perfil del paciente con osteoporosis que acude a laconsulta de Medicina Interna en Espa˜na. Además, ponen de manifiesto el carácter multisistémicode esta entidad que junto con su elevada prevalencia determinan que las consultas específicasde Medicina Interna dedicadas a su manejo son el lugar adecuado para la atención de estos pacientes

Preserved antigenicity of HIV-1 p24 produced and purified in high yields from plants inoculated with a tobacco mosaic virus (TMV)-derived vector

Production of structural proteins from foot-and-mouth disease virus (FMDV) and bovine herpes virus (BHV-1) in Nicotiana benthamiana through the use of a tobacco mosaic virus-based vector (TMV-30B) has been reported previously. The development of the TMV-30B-HISc vector, a new version that adds a C-terminal histidine (His) sequence to the foreign protein expressed is described. Coding sequences from the FMDV VPl protein and the core protein, p24, from a clade C HIV-1 isolate from Zambia were cloned into the new vector and infective RNAs were generated for each construct to inoculate N. benthamiana plants. His-tagged proteins were purified from inoculated leaves using immobilized metal affinity chromatography (IMAC) as detected by Coomassie blue staining and proteins were further characterized in Western blot assays using a commercial anti-6xHis mAb and specific polyclonal antisera for each protein. While yields obtained for the VPl-His protein after purification were similar to those in crude extracts obtained with the previous TMV-VPl vector, p24-His yields were 10–15 times higher than those of VPl-His. Twenty-five grams of TMV-p24-HISc inoculated leaves were processed to obtain 2.5 mg of isolated p24-His and the recombinant protein was inoculated in rabbits to test immunogenicity and antigenic integrity of the plant-produced p24-His. Animals developed a strong and specific humoral response to the p24-His after the first booster and immune sera was able to recognize the native p24 from a different clade expressed on the surface of the HIV-1 chronically infected HUT78/ARV T-cell line. Importantly, the recombinant p24-His proved its efficiency by confirming the serology of 117 samples previously tested by two rapid HIV-1 tests, thus representing an excellent alternative for production of highly specific diagnostic reagents for HIV endemic regions in the developing world

Multimerization of peptide antigens for production of stable immunogens in transgenic plants

Previous literature addressing the production of recombinant proteins in heterologous systems has consistently shown that proteins capable of forming complex structures tend to accumulate within host cells at relatively higher levels than monomeric forms. In this report, we translationally fused a 21-aminoacids long highly immunogenic peptide (2L21), derived from canine parvovirus (CPV) VP2 protein to a 41-aminoacid long tetramerization domain (TD) from the transcriptional factor p53. The chimerical DNA construction 2L21-TD was cloned in a binary plant transformation vector and used to transform Arabidopsis thaliana plants. Fifteen of the 25 transgenic lines obtained in the experiment showed detectable 2L21-TD RNA accumulation and from these we chose 4 to study 2L21-TD protein accumulation. Non-denaturing immunoblotting assays revealed that 2L21-TD chimeras effectively formed tetrameric complexes with yields reaching up to 12 μg/mg of soluble protein. Mice immunized by oral or intraperitoneal routes with crude protein extracts containing 2L21-TD protein were able to detect both 2L21-synthetic peptide and CPV VP2 proteins, with titers similar to those elicited by a previously reported fusion between 2L21 and the β-glucuronidase protein. These results demonstrate that multimerization directed by the small TD domain contributed to the stabilization and consequently to the accumulation of the 2L21 peptide in transgenic plants, without altering its native antigenicity and immunogenicity. © 2006 Elsevier B.V. All rights reserved