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Publications

The Pirbright Institute publication directory contains details of selected publications written by our researchers.

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Abstract

Members of the family Picornaviridae consist of small positive-sense single-stranded RNA (+ssRNA) viruses capable of infecting various vertebrate species, including birds. One of the recently identified avian picornaviruses, with a remarkably long (>9,040-nucleotide) but still incompletely sequenced genome, is turkey hepatitis virus 1 (THV-1; species Melegrivirus A, genus Megrivirus), a virus associated with liver necrosis and enteritis in commercial turkeys (Meleagris gallopavo). This report presents the results of the genetic analysis of three complete genomes of megriviruses from fecal samples of chickens (chicken/B21-CHV/2012/HUN, GenBank accession no. KF961186, and chicken/CHK-IV-CHV/2013/HUN, GenBank accession no. KF961187) (Gallus gallus domesticus) and turkey (turkey/B407-THV/2011/HUN, GenBank accession no. KF961188) (Meleagris gallopavo) with the largest picornavirus genome (up to 9,739 nucleotides) so far described. The close phylogenetic relationship to THV-1 in the nonstructural protein-coding genome region and possession of the same internal ribosomal entry site type (IVB-like) suggest that the study strains belong to the genus Megrivirus. However, the genome comparisons revealed numerous unique variations (e.g., different numbers of potential 2A peptides, unusually long 3? genome parts with various lengths of a potential second open reading frame, and multiple repeating sequence motifs in the 3? untranslated region) and heterogeneous sequence relationships between the structural and nonstructural genome regions. These differences suggest the classification of chicken megrivirus-like viruses into a candidate novel species in the genus Megrivirus. Based on the different phylogenetic positions of chicken megrivirus-like viruses at the structural and nonstructural genome regions, the recombinant nature of these viruses is plausible. The comparative genome analysis of turkey and novel chicken megriviruses revealed numerous unique genome features, e.g., up to four potential 2A peptides, unusually long 3? genome parts with various lengths containing a potential second open reading frame, multiple repeating sequence motifs, and heterogeneous sequence relationships (possibly due to a recombination event) between the structural and nonstructural genome regions. Our results could help us to better understand the evolution and diversity (in terms of sequence and genome layout) of picornaviruses.

Abstract

Recent laboratory successes in the development of genetically engineered mosquitoes for controlling pathogen transmission have fostered the need for standardized procedures for advancing the technical achievements to practical tools. It is incumbent in many cases for the same scientists doing the in-laboratory discovery research to also take on the initial challenges of developing the pathway that will move the technologies to the field. One of these challenges is having a set of criteria for selecting collaborators and sites for efficacy and safety field trials that combine rigorous science with good ethical and legal practices. Specific site-selection criteria were developed in four categories-Scientific, Regulatory, Community Engagement, and Resources-in anticipation of open-field releases of a transgenic mosquito strain designed to suppress populations of the dengue vector mosquito, Aedes aegypti. The criteria are derived from previous published material, discussions, and personal experiences with the expectation of providing guidance to laboratory scientists for addressing the conceptual and operational considerations for identifying partner researchers and countries with whom to collaborate. These criteria are not intended to be prescriptive nor can they be applied to every circumstance where genetic approaches are proposed for deployment. However, we encourage those involved in the discovery phase of research to consider each criterion during project planning activities, and where appropriate, incorporate them into a "go/no-go" decision-making process for further development and testing of the technologies.
Browning H M, Acevedo-Whitehouse K, Gulland F M D, Hall A J, Finlayson J, Dagleish M P, Billington K J, Colegrove K, Hammond J A (2014)

Evidence for a genetic basis of urogenital carcinoma in the wild California sea lion

Proceedings of the Royal Society B 281 (1796), 20140240

Abstract

Although neoplasia is a major cause of mortality in humans and domestic animals, it has rarely been described in wildlife species. One of the few examples is a highly prevalent urogenital carcinoma in California sea lions (CSLs). Although the aetiology of this carcinoma is clearly multifactorial, inbreeding depression, as estimated using levels of microsatellite multilocus heterozygosity, is identified as predictive for this neoplasia. On further analysis, this relationship appears to be largely driven by one marker, suggesting that a single locus might be associated with the occurrence of this disease in CSLs. In a case–control study, carcinoma was significantly associated with homozygosity at the Pv11 microsatellite locus. Pv11 was mapped to intron 9 of the heparanase 2 gene (HPSE2) locus, a very large gene encoding heparanase 2, which in humans is associated with multiple carcinomas. Correspondingly, immunohistochemical labelling in tissues was present in carcinoma cases within a single homozygous Pv11 genotype. To our knowledge, this is the first report of an individual locus being associated with cancer in any wildlife species. This adds emphasis to the study of HPSE2 in other species, including humans and will guide future studies on this sentinel species that shares much of its diet and environment with humans

Abstract

The impact of morbilliviruses on both human and animal populations is well documented in the history of mankind. Indeed, prior to the development of vaccines for these diseases, morbilliviruses plagued both humans and their livestock that were heavily relied upon for food and motor power within communities. Measles virus (MeV) was responsible for the death of millions of people annually across the world and those fortunate enough to escape the disease often faced starvation where their livestock had died following infection with rinderpest virus (RPV) or peste des petits ruminants virus (PPRV). Canine distemper virus has affected dog populations for centuries and in the past few decades appears to have jumped species, now causing disease in a number of non-canid species, some of which are been pushed to the brink of extinction by the virus. During the age of vaccination, the introduction and successful application of vaccines against rinderpest and measles has led to the eradication of the former and the greater control of the latter. Vaccines against PPR and canine distemper have also been generated; however, the diseases still pose a threat to susceptible species. Here we review the currently available vaccines against these four morbilliviruses and discuss the prospects for the development of new generation vaccines.
Calvo-Pinilla E, Castillo-Olivares J, Jabbar T, Ortego J, de la Poza F, Marín-López A (2014)

Recombinant vaccines against bluetongue virus

Virus Research 182 (0), 78-86

Abstract

Bluetongue (BT) is a hemorrhagic disease of ruminants caused by bluetongue virus (BTV), the prototype member of the genus Orbivirus within the family Reoviridae and is transmitted via biting midges of the genus Culicoides. BTV can be found on all continents except Antarctica, and up to 26 immunologically distinct BTV serotypes have been identified. Live attenuated and inactivated BTV vaccines have been used over the years with different degrees of success. The multiple outbreaks of BTV in Mediterranean Europe in the last two decades and the incursion of BTV-8 in Northern Europe in 2008 has re-stimulated the interest to develop improved vaccination strategies against BTV. In particular, safer, cross-reactive, more efficacious vaccines with differential diagnostic capability have been pursued by multiple BTV research groups and vaccine manufacturers. A wide variety of recombinant BTV vaccine prototypes have been investigated, ranging from baculovirus-expressed sub-unit vaccines to the use of live viral vectors. This article gives a brief overview of all these modern approaches to develop vaccines against BTV including some recent unpublished data.

Abstract

In previous studies we showed that a recombinant Modified Vaccinia Ankara (MVA) virus expressing the protein VP2 of AHSV serotype 4 (MVA-VP2) induced virus neutralising antibodies in horses and protected interferon alpha receptor gene knock-out mice (IFNAR?/?) against challenge. We continued these studies and determined, in the IFNAR?/? mouse model, whether the antibody responses induced by MVA-VP2 vaccination play a key role in protection against AHSV. Thus, groups of mice were vaccinated with wild type MVA (MVA-wt) or MVA-VP2 and the antisera from these mice were used in a passive immunisation experiment. Donor antisera from (a) MVA-wt; (b) MVA-VP2 vaccinated; or (c) MVA-VP2 vaccinated and AHSV infected mice, were transferred to AHSV non-immune recipient mice. The recipients were challenged with virulent AHSV together with MVA-VP2 vaccinated and MVA-wt vaccinated control animals and the levels of protection against AHSV-4 were compared between all these groups. The results showed that following AHSV challenge, mice that were passively immunised with MVA-VP2 vaccinated antisera were highly protected against AHSV disease and had lower levels of viraemia than recipients of MVA-wt antisera. Our study indicates that MVA-VP2 vaccination induces a highly protective humoral immune response against AHSV.
Caporale M, Di Gialleonorado L, Janowicz A, Wilkie G, Shaw A, Savini G, Van Rijn P A, Mertens P, Di Ventura M, Palmarini M (2014)

Virus and host factors affecting the clinical outcome of bluetongue virus infection

Journal of Virology 88 (18), 10399-10411

Abstract

Bluetongue is a major infectious disease of ruminants caused by bluetongue virus (BTV), an arbovirus transmitted by Culicoides. Here, we assessed virus and host factors influencing the clinical outcome of BTV infection using a single experimental framework. We investigated how mammalian host species, breed, age, BTV serotypes, and strains within a serotype, affect the clinical course of bluetongue. Results obtained indicate that in small ruminants there is a marked difference in the susceptibility to clinical disease induced by BTV at the host species level, but less so at the breed level. No major differences in virulence were found between divergent serotypes (BTV-8 and BTV-2). However, we observed striking differences in virulence between closely related strains of the same serotype collected towards the beginning and the end of the European BTV-8 outbreak. As observed previously, differences in disease severity were also observed when animals were infected with either blood from a BTV-infected animal or from the same virus isolated in cell culture. Interestingly, with the exception of two silent mutations, full viral genome sequencing showed identical consensus sequences of the virus before and after cell culture isolation. However, deep sequencing analysis revealed a marked decrease in the genetic diversity of the viral population after passaging in mammalian cells. In contrast, passaging in Culicoides cells increased the overall number of low frequency variants compared to virus never passaged in cell culture. Thus, Culicoides might be a source of new viral variants and viral population diversity can be another factor influencing BTV virulence.IMPORTANCE Bluetongue is one of the major infectious diseases of ruminants. It is caused by an arbovirus known as Bluetongue virus (BTV). The clinical outcome of BTV infection is extremely variable. We show that there are clear links between the severity of bluetongue and the mammalian host species infected, while at the breed level differences were less evident. No differences were observed in the virulence of two different BTV serotypes (BTV-8 and BTV-2). In contrast, we show that the European BTV-8 strain isolated at the beginning of the bluetongue outbreak in 2006 was more virulent than a strain isolated towards the end of the outbreak. In addition, we show that there is a link between the variability of the BTV population as a whole and virulence and our data also suggest that Culicoides cells might function as an "incubator" of viral variants.

Abstract

Following the incursions into the UK of bluetongue virus (BTV) in 2007 and Schmallenberg virus (SBV) in 2011, an increasingly pertinent question for those tasked with predicting and responding to outbreaks of livestock arboviruses in the UK is, what is next? Culicoides, the biting midges that transmit BTV and SBV, also act as vectors for over 50 described arboviruses worldwide, although we currently know very little about the vast majority of these, even in the countries in which they were originally identified. In addition, it is likely there are a large number of previously unidentified Culicoides-borne arboviruses that have the potential to arrive in northern Europe with little warning (as in the case of SBV). While unidentified viruses pose serious challenges for the prediction of epidemiology and pathogenicity (Carpenter and others 2013), perhaps the greatest concern in Europe currently surrounds the potential for the emergence of a well-known Culicoides-borne arbovirus, African horse sickness virus (AHSV).
Casey M B, Lembo T, Knowles N J, Fyumagwa R, Kivaria F, Maliti H, Kasanga C, Sallu R, Reeve R, Parida S, King D P, Cleaveland S (2014)

Patterns of foot-and-mouth disease virus distribution in Africa: the role of livestock and wildlife in virus emergence

The Role of Animals in Emerging Viral Diseases (edited by N Johnson, Academic Press), 21-38

Abstract

Foot-and-mouth disease (FMD) is one of the most highly contagious diseases of animals. The disease is distributed on three continents (Asia, Africa and South America) where it disrupts the food security of people who depend on livestock and animal products. Substantial economic losses are associated with controlling FMD outbreaks in many countries. For example, during the epidemic in the UK in 2001, losses to agriculture were estimated to be £3.1 billion, with similar losses arising from negative impacts on tourism. Over 4 million animals were slaughtered as part of FMD control measures and a further 2 million were slaughtered due to welfare issues associated with animal movement bans. FMD also has a devastating impact on rural livelihoods and livestock trade opportunities in developing countries where it is endemic. In Africa, historic patterns of FMD virus (FMDV) emergence are likely to have been shaped by the introduction and subsequent eradication of rinderpest. However, important questions remain about contemporary drivers of disease distribution. In particular, the relative contribution of wildlife as sources of infection for livestock and factors affecting the potential for cross-species transmission, and mechanisms of viral maintenance in endemic regions are poorly understood. These issues encompass a complex suite of interacting social, ecological and economic factors that act as drivers of change in patterns of land-use, livestock movements, international trade, and conservation of wildlife-protected areas. Understanding patterns of FMDV infection at the livestock-wildlife interface is of particular importance for designing and developing appropriate disease control strategies in sub-Saharan Africa, and of increasing interest as momentum grows for global control of FMD within the framework of the Progressive Control Pathway (PCP-FMD). This chapter reviews the historical distribution and emergence of FMDV in Africa, and factors that govern the current circulation and maintenance of FMDV in sub-Saharan Africa.

Abstract

We aimed to assess the status of naturally occurring CD4(+) and CD8(+) T cell responses to a tumour associated antigen, Mesothelin, in patients with pancreatic carcinoma and study the effects of elevated IL-10 on Mesothelin-specific T cell responses. For that sake, short term T cell lines were generated from PBMCs of 16 healthy controls, 15 patients with benign pancreatic diseases and 25 patients with pancreatic carcinoma and Mesothelin-specific CD4(+) and CD8(+) T cell responses were analysed using intracellular cytokine assays for IFN-gamma. Plasma levels of IL-10 and Mesothelin were measured using cytometric bead array and ELISA assay, respectively. The blocking assays were performed to assess the effects of IL-10 on Mesothelin-specific T cell responses. Here, we demonstrate that the plasma levels of Mesothelin and IL-10 are significantly increased in patients with pancreatic carcinoma. Additionally, we found that (a) Mesothelin-specific T cell responses are significantly expanded in cancer patients (p = 0.0053), (b) the multifunctional CD4(+) T cell response is directed toward a broad repertoire of epitopes within the Mesothelin protein. (c) Mesothelin-specific CD4+ T cell response is directly inhibited by elevated IL-10 in cancer patients. These data provides evidence for the use of Mesothelin as an immunogen for tumourspecific T cell response.

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