Co-expression of VP2, NS1 and NS2-Nt proteins by an MVA viral vector induces complete protection against bluetongue virus.

Introduction: Bluetongue (BT), caused by bluetongue virus (BTV), is an important arthropod-borne livestock disease listed by the World Organization for Animal Health. Live-attenuated and inactivated vaccines have permitted to control BT but they do not simultaneously protect against the myriad of BTV serotypes. Recently, we identified the highly conserved BTV nonstructural protein NS1 and the N-terminal region of NS2 as antigens capable of conferring multiserotype protection against BTV. Methods: Here, we designed Modified Vaccinia Ankara (MVA) viral vectors that expressed BTV-4 proteins VP2 or VP7 along with NS1 and NS2-Nt as well as MVAs that expressed proteins VP2, VP7 or NS1 and NS2-Nt. Results: Immunization of IFNAR(-/-) mice with two doses of MVA-NS1-2A-NS2-Nt protected mice from BTV-4M infection by the induction of an antigen-specific T cell immune response. Despite rMVA expressing VP7 alone were not protective in the IFNAR(-/-) mouse model, inclusion of VP7 in the vaccine formulation amplified the cell-mediated response induced by NS1 and NS2-Nt. Expression of VP2 elicited protective non-cross-reactive neutralizing antibodies (nAbs) in immunized animals and improved the protection observed in the MVA-NS1-2A-NS2-Nt immunized mice when these three BTV antigens were co-expressed. Moreover, vaccines candidates co-expressing VP2 or VP7 along with NS1 and NS2-Nt provided multiserotype protection. We assessed protective efficacy of both vaccine candidates in sheep against virulent challenge with BTV-4M. Discussion: Immunization with MVA-VP7-NS1-2A-NS2-Nt partially dumped viral replication and clinical disease whereas administration of MVA-VP2-NS1-2A-NS2-Nt promoted a complete protection, preventing viraemia and the pathology produced by BTV infection.

IFNAR(-/-) Mice Constitute a Suitable Animal Model for Epizootic Hemorrhagic Disease Virus Study and Vaccine Evaluation.

Epizootic hemorrhagic disease (EHD), caused by Epizootic hemorrhagic disease virus (EHDV), is an emerging and severe livestock disease. Recent incursion and distribution of EHDV in Europe have outlined the emerging character of EHD. Despite its worldwide impact, numerous knowledge gaps exist. A range of inconveniences restricts utilization of natural hosts of EHDV. Here, we show that adult mice deficient in type I IFN receptor (IFNAR(-/-)) are highly susceptible to EHDV-6 and EHDV-8 infection when the virus is administered subcutaneously. Disease was characterized by ruffled hair, reluctance to move, dehydration and conjunctivitis, with viraemia detected from day 5 post-infection. A deeper characterization of EHDV-8 infection showed viral replication in the lung, liver, spleen, kidney, testis and ovaries. Importantly, increased expression levels of pro-inflammatory cytokines IL-1ß, IL-6 and CXCL2 were observed in spleen after EHDV-8 infection. Furthermore, IFNAR(-/-) adult mice immunized with a EHDV-8 inactivated vaccine elicited neutralizing antibodies specific of EHDV-8 and full protection against challenge with a lethal dose of this virus. This study also explores the possibilities of this animal model for study of BTV and EHDV coinfection. In summary, the IFNAR(-/-) mouse model faithfully recapitulates EHD and can be applied for vaccine testing, which can facilitate progress in addressing the animal health challenge posed by this virus.

Novel Reference Method for the Characterization of PD Measuring Systems Using HFCT Sensors.

During their lifespan, high-voltage (HV) electrical systems are subjected to operating conditions in which electrical, mechanical, thermal and environmental-related stresses occur. These conditions over time lead to unforeseen failures caused by various types of defects. For this reason, there are several technologies for measuring and monitoring the electrical systems, with the aim of minimizing the number of faults. The early detection of defects, preferably in their incipient state, will enable the necessary corrective actions to be taken in order to avoid unforeseen failures. These failures generally lead to human risks and material damage, lack of power supply and significant economic losses. An efficient maintenance technique for the early detection of defects consists of the supervision of the dielectrics status in the installations by means of on-line partial discharge (PD) measurement. Nowadays, there are numerous systems in the market for the measurement of PD in HV installations. The most efficient with a reasonable cost will be those that offer greater security guarantees and the best positioned in the market. Currently, technology developers and users of PD measuring systems face difficulties related to the lack of reference procedures for their complete characterization and to the technical and economic drawback of performing the characterization tests on site or in laboratory installations. To deal with the previous difficulties, in this paper a novel method for the complete and standardized characterization of PD measuring systems is presented. The applicability of this method is mainly adapted for the characterization of systems operating in on-line applications using high-frequency current transformer (HFCT) sensors. For the appropriate application of the method, an associated and necessary scale modular test platform is used. In the test platform, the real on-site measuring conditions of an HV insulated distribution line are simulated in a controlled way. Practical characterizations, showing the convenience and advantages of applying the method using the modular test platform, are also presented.

Engineering recombinant replication-competent bluetongue viruses expressing reporter genes for in vitro and non-invasive in vivo studies.

Bluetongue virus (BTV) is the causative agent of the important livestock disease bluetongue (BT), which is transmitted via Culicoides bites. BT causes severe economic losses associated with its considerable impact on health and trade of animals. By reverse genetics, we have designed and rescued reporter-expressing recombinant (r)BTV expressing NanoLuc luciferase (NLuc) or Venus fluorescent protein. To generate these viruses, we custom synthesized a modified viral segment 5 encoding NS1 protein with the reporter genes located downstream and linked by the Porcine teschovirus-1 (PTV-1) 2A autoproteolytic cleavage site. Therefore, fluorescent signal or luciferase activity is only detected after virus replication and expression of non-structural proteins. Fluorescence or luminescence signals were detected in cells infected with rBTV/Venus or rBTV/NLuc, respectively. Moreover, the marking of NS2 protein confirmed that reporter genes were only expressed in BTV-infected cells. Growth kinetics of rBTV/NLuc and rBTV/Venus in Vero cells showed replication rates similar to those of wild-type and rBTV. Infectivity studies of these recombinant viruses in IFNAR(-/-) mice showed a higher lethal dose for rBTV/NLuc and rBTV/Venus than for rBTV indicating that viruses expressing the reporter genes are attenuated in vivo. Interestingly, luciferase activity was detected in the plasma of viraemic mice infected with rBTV/NLuc. Furthermore, luciferase activity quantitatively correlated with RNAemia levels of infected mice throughout the infection. In addition, we have investigated the in vivo replication and dissemination of BTV in IFNAR (-/-) mice using BTV/NLuc and non-invasive in vivo imaging systems.IMPORTANCEThe use of replication-competent viruses that encode a traceable fluorescent or luciferase reporter protein has significantly contributed to the in vitro and in vivo study of viral infections and the development of novel therapeutic approaches. In this work, we have generated rBTV that express fluorescent or luminescence proteins to track BTV infection both in vitro and in vivo. Despite the availability of vaccines, BTV and other related orbivirus are still associated with a significant impact on animal health and have important economic consequences worldwide. Our studies may contribute to the advance in orbivirus research and pave the way for the rapid development of new treatments, including vaccines.

Cytokine mRNA Expression Profile in Target Organs of IFNAR (-/-) Mice Infected with African Horse Sickness Virus.

African horse sickness (AHS) is a highly severe disease caused by a viral etiological agent, African horse sickness virus (AHSV). It is endemic in sub-Saharan Africa, while sporadic outbreaks have occurred in North Africa, Asia, and Europe, with the most recent cases in Thailand. AHSV transmission between equines occurs primarily by biting midges of the genus Culicoides, especially C. imicola, with a wide distribution globally. As research in horses is highly restricted due to a variety of factors, small laboratory animal models that reproduce clinical signs and pathology observed in natural infection of AHSV are highly needed. Here, we investigated the expression profile of several pro-inflammatory cytokines in target organs and serum of IFNAR (-/-) mice, to continue characterizing this established animal model and to go deep into the innate immune responses that are still needed.

Afectación subgaleal secundaria del linfoma de células del manto / Secondary presentation of mantle cell lymphoma in the subgaleal layer of the scalp

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Osteoartropatía hipertrófica: una rara manifestación de un tumor amigdalar / Hypertrophic osteoarthropathy: a rare manifestation of an amygdalar tumour

Presentamos un caso de osteoartropatía hipertrófica como manifestación acompañante de un tumor maligno amigdalar, en un paciente al que se detectó la anomalía ósea 10 meses antes de la presentación clínica del tumor. La osteoartropatía hipertrófica pneúmica es un síndrome clínico-radiológico caracterizado por proliferación del periostio de huesos tubulares con hipocratismo digital. En más del 90 % de los casos se ha relacionado con un tumor intratorácico, más raramente secundario a hemopatía maligna o cáncer otorrinolaríngeo, sobre todo de rinofaringe. La asociación con tumor amigdalar, en nuestro conocimiento, no se ha descrito

A case of hypertrophic osteoarthropathy in association with malignant tonsil tumour is reported. It involved a patient in whom the bone disorder was detected 10 months prior to the clinical manifestation of the tumour. Hypertrophic pneumic osteoarthropathy is a clinical and radiological syndrome characterized by proliferation of periosteum in long bones with digital clubbing. In over 90 % of the cases, it has been associated with an intra-thoracic tumour and more rarely secondary to a malignant haemopathy or ENT cancer, particularly of the rhinopharynx. As far as we know, the association with carcinoma of the tonsils has not previously been described