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Peredo E.L.,University of Oviedo | Revilla M.A.,University of Oviedo | Reed B.M.,U.S. Department of Agriculture | Javornik B.,University of Ljubljana | And 3 more authors.
Genetic Resources and Crop Evolution | Year: 2010

Microsatellite variation at the nuclear and chloroplast genomes was evaluated for wild European and wild American hops, in order to assess the genetic diversity and origin of cultivated hops. Seven nuclear loci and 32 chloroplast loci were used in the analysis of 182 hop accessions including wild European (68), wild American (48), and cultivars (66). A total of 116 alleles were identified using 7 nuclear microsatellites showing different averages of polymorphism and distribution in the wild American and European accessions and cultivars. Two main groups were established as revealed by several statistical analyses; one including European wild accessions and cultivars and a second group consisting of American wild accessions. Three polymorphic chloroplast microsatellite loci were detected, six alleles were scored which defined a total of five haplotypes that were exclusive or presented different distribution between American and European wild accessions. A major influence of the wild European haplotypes was detected among hop cultivars. To the best of our knowledge, this is the first work reporting the use of chloroplast microsatellites in hops. © 2009 Springer Science+Business Media B.V.


Qi Z.,Yancheng Institute of Technology | Qi Z.,University of Aberdeen | Zhang Q.,Yancheng Institute of Technology | Holland J.W.,University of Aberdeen | And 4 more authors.
Fisheries Science | Year: 2016

Chemokine-like receptors (CMKLRs) are multi-functional receptors with roles in regulating leukocyte and inflammation. Currently, two members of the CMKLRs, CMKLR1 and CMKLR2, have been found in salmonids, indicating that the CMKLRs had expanded from an ancestor gene in teleost fish. In the present study, the third member of the CMKLRs, defined as CMKLR3, was identified and cloned in rainbow trout. The trout CMKLR3 possessed conserved features of the CMKLR family including seven transmembrane regions, a dynein regulatory complex (DRC) motif, and two cysteine residues, but shared low sequence identities with fish CMKLR1 and CMKLR2 (24‒38 %), which was confirmed by phylogenetic tree analysis. Trout CMKLR3 was highly expressed in body kidney, head kidney, and IgM+ B cells, indicating its functional role in regulating leukocytes. We were able to modulate the expression of trout CMKLR3 in vivo by bacterial and parasitic infections but it remained apathetic to virus infection, and it was also successfully modulated in vitro by peptidoglycan and cytokines (IFN-γ and IL-6). Our results suggest that trout CMKLR3 is regulated in a complex way and has an important regulatory role in inflammatory responses. © 2016 Japanese Society of Fisheries Science


Martin V.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Pascual E.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Avia M.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Pena L.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | And 2 more authors.
PLoS ONE | Year: 2015

Bluetongue virus (BTV) is an economically important Orbivirus of the Reoviridae family that causes a hemorrhagic disease in ruminants. Its control has been achieved by inactivatedvaccines that have proven to protect against homologous BTV challenge although unable to induce long-term immunity. Therefore, a more efficient control strategy needs to be developed. Recombinant adenovirus vectors are lead vaccine candidates for protection of several diseases, mainly because of their potency to induce potent T cell immunity. Here we report the induction of humoral and T-cell mediated responses able to protect animals against BTV challenge by recombinant replication-defective human adenovirus serotype 5 (Ad5) expressing either VP7, VP2 or NS3 BTV proteins. First we used the IFNAR(-/-) mouse model system to establish a proof of principle, and afterwards we assayed the protective efficacy in sheep, the natural host of BTV. Mice were completely protected against BTV challenge, developing humoral and BTV-specific CD8+- and CD4+-T cell responses by vaccination with the different rAd5. Sheep vaccinated with Ad5-BTV-VP2 and Ad5-BTV-VP7 or only with Ad5-BTV-VP7 and challenged with BTV showed mild disease symptoms and reduced viremia. This partial protection was achieved in the absence of neutralizing antibodies but strong BTV-specific CD8+ T cell responses in those sheep vaccinated with Ad5- BTV-VP7. These data indicate that rAd5 is a suitable vaccine vector to induce T cell immunity during BTV vaccination and provide new data regarding the relevance of T cell responses in protection during BTV infection. © 2015 Martín et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Rojas J.-M.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Rodriguez-Calvo T.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Pena L.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Sevilla N.,Instituto Nacional Of Investigacion Agraria Y Alimentaria
Vaccine | Year: 2011

Bluetongue virus (BTV), an economically important orbivirus of the Reoviridae family, is a non-enveloped, dsRNA virus that causes a haemorrhagic disease mainly in sheep, but little is known of the cellular immunity elicited against BTV. We observed that vaccination of interferon type I-deficient mice (IFNAR (-/-)), or inoculation of the wild type C57BL/6 strain with BTV-8, induced a strong T cell response. Therefore, we proceeded to identify some of the T cell epitopes targeted by the immune system. We selected, using H-2 b-binding predictive algorithms, 3 major histocompatibility complex (MHC)-class II-binding peptides and 7 MHC-class I binding peptides from the BTV-8 core protein VP7, as potential T cell epitopes. Peptide binding assays confirmed that all 7 MHC-class I predicted peptides bound MHC-class I molecules. Three MHC-class I and 2 MHC-class II binding peptide consistently elicited peptide-specific IFN-γ production (as measured by ELISPOT assays) in splenocytes from C57BL/6 BTV-8-inoculated mice and IFNAR (-/-)-vaccinated mice. The functionality of these T cells was confirmed by proliferation and cytotoxicity assays. Flow cytometry analysis demonstrated that CD8 + T cells responded to MHC-class I binding peptides and CD4 + T cells to MHC-class II binding peptides. Importantly, these 5 epitopes were also able to induced IFN-γ production in sheep inoculated with BTV-8. Taken together, these data demonstrate the activation of BTV-specific T cells during infection and vaccination. The characterisation of these novel T cell epitopes may also provide an opportunity to develop DIVA-compliant vaccination approach to BTV encompassing a broad-spectrum of serotypes. © 2011 Elsevier Ltd.


Warimwe G.M.,University of Oxford | Lorenzo G.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Lopez-Gil E.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Reyes-Sandoval A.,University of Oxford | And 11 more authors.
Virology Journal | Year: 2013

Background: Rift Valley Fever (RVF) is a viral zoonosis that historically affects livestock production and human health in sub-Saharan Africa, though epizootics have also occurred in the Arabian Peninsula. Whilst an effective live-attenuated vaccine is available for livestock, there is currently no licensed human RVF vaccine. Replication-deficient chimpanzee adenovirus (ChAd) vectors are an ideal platform for development of a human RVF vaccine, given the low prevalence of neutralizing antibodies against them in the human population, and their excellent safety and immunogenicity profile in human clinical trials of vaccines against a wide range of pathogens. Methods. Here, in BALB/c mice, we evaluated the immunogenicity and efficacy of a replication-deficient chimpanzee adenovirus vector, ChAdOx1, encoding the RVF virus envelope glycoproteins, Gn and Gc, which are targets of virus neutralizing antibodies. The ChAdOx1-GnGc vaccine was assessed in comparison to a replication-deficient human adenovirus type 5 vector encoding Gn and Gc (HAdV5-GnGc), a strategy previously shown to confer protective immunity against RVF in mice. Results: A single immunization with either of the vaccines conferred protection against RVF virus challenge eight weeks post-immunization. Both vaccines elicited RVF virus neutralizing antibody and a robust CD8§ssup§+§esup§ T cell response. Conclusions: Together the results support further development of RVF vaccines based on replication-deficient adenovirus vectors, with ChAdOx1-GnGc being a potential candidate for use in future human clinical trials. © 2013 Warimwe et al.; licensee BioMed Central Ltd.


Santin-Montanya M.I.,Instituto Nacional Of Investigacion Agraria Y Alimentaria | Jimenez J.,TRAGSA | Ocana L.,TRAGSA | Sanchez F.J.,Direccion General del Agua
Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes | Year: 2013

Chlorophyll fluorescence analysis (CFA) has been successfully used to rapidly determine the responses of different plants to herbicides. It has not, however, been used to test the effect of these products on invasive riparian species. This paper reports the use of CFA to determine photosynthetic activity in Arundo donax, an invasive reed causing serious problems in Mediterranean riparian habitats, in response to systemic herbicide application following cutting. Growth was measured in terms of new sprout relative height and sprout and rhizome relative biomass. CFA showed glyphosate, from half the on-label dose of 5 L ai.ha-1upwards, to have a significant effect (100% reduction) on photosynthetic activity at 21 days after treatment (DAT), while profoxydim, from half the on-label dose of 0.375 L ai.ha-1upwards, caused a 70% reduction soon after application, although these plants later recovered. Azimsulfuron, cyhalofop-butyl and penoxsulam had no significant effect on photosynthetic activity at any dose. At 60 DAT, glyphosate (from half the on-label dose of 5 L ai.ha-1upwards) was associated with a 90% reduction in sprout height, while profoxydim (from half the on-label dose of 0.375 L ai.ha-1upwards) caused a 50% reduction. No dose (0-2x the on-label dose) of azimsulfuron, penoxsulam or cyhalofop-butyl was associated with any significant growth reduction at 60 DAT. The results show that CFA can be used to successfully measure the response of these invasive plants to herbicides, and that glyphosate, and possibly profoxydim, might be used to control Arundo donax after initial cutting. © 2013 Copyright Taylor and Francis Group, LLC.


PubMed | Instituto Nacional Of Investigacion Agraria Y Alimentaria
Type: Journal Article | Journal: Journal of virology | Year: 2013

Hematopoietic stem cells (HSCs) give rise to progenitors with potential to produce multiple cell types, including dendritic cells (DCs). DCs are the principal antigen-presenting cells and represent the crucial link between innate and adaptive immune responses. Bluetongue virus (BTV), an economically important Orbivirus of the Reoviridae family, causes a hemorrhagic disease mainly in sheep and occasionally in other species of ruminants. BTV is transmitted between its mammalian hosts by certain species of biting midges (Culicoides spp.) and is a potent alpha interferon (IFN-) inducer. In the present report, we show that BTV infects cells of hematopoietic origin but not HSCs in immunocompetent sheep. However, BTV infects HSCs in the absence of type I IFN (IFN-I) signaling in vitro and in vivo. Infection of HSCs in vitro results in cellular death by apoptosis. Furthermore, BTV infects bone marrow-derived DCs (BM-DCs), interfering with their development to mature DCs in the absence of type I IFN signaling. Costimulatory molecules CD80 and CD86 and costimulatory molecules CD40 and major histocompatibility complex class II (MHC-II) are affected by BTV infection, suggesting that BTV interferes with DC antigen-presenting capacity. In vivo, different DC populations are also affected during the course of infection, probably as a result of a direct effect of BTV replication in DCs and the production of infectious virus. These new findings suggest that BTV infection of HSCs and DCs can impair the immune response, leading to persistence or animal death, and that this relies on IFN-I.


PubMed | Instituto Nacional Of Investigacion Agraria Y Alimentaria
Type: Journal Article | Journal: PloS one | Year: 2015

Bluetongue virus (BTV) is an economically important Orbivirus of the Reoviridae family that causes a hemorrhagic disease in ruminants. Its control has been achieved by inactivated-vaccines that have proven to protect against homologous BTV challenge although unable to induce long-term immunity. Therefore, a more efficient control strategy needs to be developed. Recombinant adenovirus vectors are lead vaccine candidates for protection of several diseases, mainly because of their potency to induce potent T cell immunity. Here we report the induction of humoral and T-cell mediated responses able to protect animals against BTV challenge by recombinant replication-defective human adenovirus serotype 5 (Ad5) expressing either VP7, VP2 or NS3 BTV proteins. First we used the IFNAR(-/-) mouse model system to establish a proof of principle, and afterwards we assayed the protective efficacy in sheep, the natural host of BTV. Mice were completely protected against BTV challenge, developing humoral and BTV-specific CD8+- and CD4+-T cell responses by vaccination with the different rAd5. Sheep vaccinated with Ad5-BTV-VP2 and Ad5-BTV-VP7 or only with Ad5-BTV-VP7 and challenged with BTV showed mild disease symptoms and reduced viremia. This partial protection was achieved in the absence of neutralizing antibodies but strong BTV-specific CD8+ T cell responses in those sheep vaccinated with Ad5-BTV-VP7. These data indicate that rAd5 is a suitable vaccine vector to induce T cell immunity during BTV vaccination and provide new data regarding the relevance of T cell responses in protection during BTV infection.


PubMed | Instituto Nacional Of Investigacion Agraria Y Alimentaria
Type: Journal Article | Journal: Vaccine | Year: 2011

Bluetongue virus (BTV), an economically important orbivirus of the Reoviridae family, is a non-enveloped, dsRNA virus that causes a haemorrhagic disease mainly in sheep, but little is known of the cellular immunity elicited against BTV. We observed that vaccination of interferon type I-deficient mice (IFNAR((-/-))), or inoculation of the wild type C57BL/6 strain with BTV-8, induced a strong T cell response. Therefore, we proceeded to identify some of the T cell epitopes targeted by the immune system. We selected, using H-2(b)-binding predictive algorithms, 3 major histocompatibility complex (MHC)-class II-binding peptides and 7 MHC-class I binding peptides from the BTV-8 core protein VP7, as potential T cell epitopes. Peptide binding assays confirmed that all 7 MHC-class I predicted peptides bound MHC-class I molecules. Three MHC-class I and 2 MHC-class II binding peptide consistently elicited peptide-specific IFN- production (as measured by ELISPOT assays) in splenocytes from C57BL/6 BTV-8-inoculated mice and IFNAR((-/-))-vaccinated mice. The functionality of these T cells was confirmed by proliferation and cytotoxicity assays. Flow cytometry analysis demonstrated that CD8(+) T cells responded to MHC-class I binding peptides and CD4(+) T cells to MHC-class II binding peptides. Importantly, these 5 epitopes were also able to induced IFN- production in sheep inoculated with BTV-8. Taken together, these data demonstrate the activation of BTV-specific T cells during infection and vaccination. The characterisation of these novel T cell epitopes may also provide an opportunity to develop DIVA-compliant vaccination approach to BTV encompassing a broad-spectrum of serotypes.


PubMed | Instituto Nacional Of Investigacion Agraria Y Alimentaria
Type: Journal Article | Journal: Journal of immunology (Baltimore, Md. : 1950) | Year: 2014

Chemokine receptor CCR7, the receptor for both CCL19 and CCL21 chemokines, regulates the recruitment and clustering of circulating leukocytes to secondary lymphoid tissues, such as lymph nodes and Peyers patches. Even though teleost fish do not have either of these secondary lymphoid structures, we have recently reported a homolog to CCR7 in rainbow trout (Oncorhynchus mykiss). In the present work, we have studied the distribution of leukocytes bearing extracellular CCR7 in naive adult tissues by flow cytometry, observing that among the different leukocyte populations, the highest numbers of cells with membrane (mem)CCR7 were recorded in the gill (7.5 2% CCR7(+) cells). In comparison, head kidney, spleen, thymus, intestine, and peripheral blood possessed <5% CCR7(+) cells. When CCR7 was studied at early developmental stages, we detected a progressive increase in gene expression and protein CCR7 levels in the gills throughout development. Surprisingly, the majority of the CCR7(+) cells in the gills were not myeloid cells and did not express membrane CD8, IgM, nor IgT, but expressed IgD on the cell surface. In fact, most IgD(+) cells in the gills expressed CCR7. Intriguingly, the IgD(+)CCR7(+) population did not coexpress memIgM. Finally, when trout were bath challenged with viral hemorrhagic septicemia virus, the number of CCR7(+) cells significantly decreased in the gills while significantly increased in head kidney. These results provide evidence of the presence of a novel memIgD(+)memIgM(-) B lymphocyte subset in trout that expresses memCCR7 and responds to viral infections. Similarities with IgD(+)IgM(-) subsets in mammals are discussed.

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