Publications

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

There were a total of 2599 results for your search.

Abstract

African swine fever virus (ASFV) causes an acute haemorrhagic disease of domestic pigs against which there is no effective vaccine. The attenuated ASFV strain OUR T88/3 has been shown previously to protect vaccinated pigs against challenge with some virulent strains including OUR T88/1. Two genes, DP71L and DP96R were deleted from the OUR T88/3 genome to create recombinant virus OUR T88/3?DP2. Deletion of these genes from virulent viruses has previously been shown to reduce ASFV virulence in domestic pigs. Groups of 6 pigs were immunised with deletion virus OUR T88/3?DP2 or parental virus OUR T88/3 and challenged with virulent OUR T88/1 virus. Four pigs (66%) were protected by inoculation with the deletion virus OUR T88/3?DP2 compared to 100% protection with the parental virus OUR T88/3. Thus the deletion of the two genes DP71L and DP96R from OUR T88/3 strain reduced its ability to protect pigs against challenge with virulent virus.
Alphey L, McKemey A, Nimmo D, Oviedo M N, Lacroix R, Matzen K, Beech C (2013)

Genetic control of Aedes mosquitoes

Pathogens and Global Health 107 (4), 170-179

Abstract

Aedes mosquitoes include important vector species such as Aedes aegypti, the major vector of dengue. Genetic control methods are being developed for several of these species, stimulated by an urgent need owing to the poor effectiveness of current methods combined with an increase in chemical pesticide resistance. In this review we discuss the various genetic strategies that have been proposed, their present status, and future prospects. We focus particularly on those methods that are already being tested in the field, including RIDL and Wolbachia-based approaches.
Arenas-Montes A J, Arenas A, Garcia-Bocanegra I, Mertens P, Batten C, Nomikou K (2013)

Serosurveillance of orbiviruses in wild cervids from Spain (Letter)

Veterinary Record 172 (19), 508-509
Publisher’s version: http://dx.doi.org/10.1136/vr.f2932

Abstract

In recent years, several emerging vectorborne diseases, including bluetongue (BTV-1, BTV-4 and BTV-8), Bagaza, Schmallenberg, Usutu and West Nile, have been reported in southern Spain, most of them originating from north Africa. Although EHDV circulation has been recently detected in livestock in the Mediterranean basin (EHDV-6 in Morocco, Algeria, Tunisia and Turkey during the period 2006-2008; EHDV-7 in Israel during 2006), no outbreaks have been reported in Europe. However, the presence of competent vectors for EHDV, the high density of wild and domestic ruminant species and the geographical proximity to north Africa, suggest that Andalusia (southern Spain) can be considered a potential risk area for the introduction of arboviruses, including EHDV. Our results suggest that EHDV has not circulated in Andalusia during the period 2006 to 2012. BTV has circulated in wild cervid populations from Andalusia in the past two years. Serological results indicate an absence of cross-reaction between EHDV and BTV in the ELISA used in the present study. EHDV in north Africa poses a major risk for European countries because of likely windborne dispersal of infected vectors. Therefore, permanent monitoring for EHDV, including active and passive surveillance in wild and domestic ruminant species and entomological surveillance, should be implemented in Andalusia.
Armesto M, Bentley K, Bickerton E, Keep S, Britton P (2013)

Coronavirus reverse genetics

Reverse Genetics of RNA Viruses: Applications and Perspectives (edited by A Bridgen, Wiley-Blackwell), 25-63
Publisher’s version:

Abstract

This chapter contains sections titled: The Coronavirinae, Infectious bronchitis, Coronavirus genome organisation, The coronavirus replication cycle, Development of reverse genetics system for coronaviruses including IBV, Reverse genetics system for IBV, Reverse genetics systems for the modification of coronavirus genomes, Using coronavirus reverse genetics systems for gene delivery

Abstract

To assess the effect of various vaccine strains on replication and shedding of virulent Marek's disease virus from experimentally infected chickens, quantitative PCR (q-PCR) methods were developed to accurately quantify viral DNA in infected chickens and in the environment in which they were housed. Four groups of 10 chickens, kept in poultry isolators, were vaccinated at 1 day old with one of four vaccines covering each of the three vaccine serotypes, then challenged with very virulent MDV strain Md5 at 8 days of age. At regular time-points, feather tips were collected from each chicken and poultry dust was collected from the air-extract prefilter of each isolator. DNA was extracted from feather and dust samples and subjected to real-time q-PCR, targeting the US2 gene of MDV-1, in order to measure Md5 level per 104 feather tip cells or per microgram of dust. Accuracy of DNA extraction from dust and real-time q-PCR were validated by comparing either q-PCR cycle threshold values or the calculated MDV genome level; for use in q-PCR, DNA was extracted from serial dilutions of MDV-infected dust diluted with noninfected dust, or DNA from MDV-infected dust was diluted with DNA from noninfected dust. The results confirmed the accuracy and sensitivity of dust DNA extraction and subsequent q-PCR and showed that differences in virus levels between dust samples truly reflect differences in shedding. Vaccination delayed both replication of Md5 in feather tips and shedding of Md5. First detection of Md5 in feather tips always preceded or coincided with first detection in dust in each group. pCVI988 and HVT+SB-1 were the most efficient vaccines in reducing both replication and shedding of Md5. There was close correlation between mean virus level in feathers of each group and mean virus level in the dust shed by that group. This relationship was similar in each of the vaccinated groups, demonstrating that measurement of the virus in dust can be used to monitor accurately both the infection status of the chickens and environmental contamination by MDV.

Abstract

The ovarian tumour (OTU) domain of the nairovirus L protein has been shown to remove ubiquitin and interferon-stimulated gene 15 protein (ISG15) from host cell proteins, which is expected to have multiple effects on cell signalling pathways. We have confirmed that the OTU domain from the L protein of the apathogenic nairovirus Dugbe virus has deubiquitinating and deISGylating activity and shown that, when expressed in cells, it is highly effective at blocking the TNF-alpha/NF-kappaB and interferon/JAK/STAT signalling pathways even at low doses. Point mutations of the catalytic site of the OTU [C40A, H151A and a double mutant] both abolished the ability of the OTU domain to deubiquitinate and deISGylate proteins and greatly reduced its effect on cell signalling pathways, confirming that it is this enzymic activity that is responsible for blocking the two signalling pathways. Expression of the inactive mutants at high levels could still block signalling, suggesting that the viral OTU can still bind to its substrate even when mutated at its catalytic site. The nairovirus L protein is a very large protein that is normally confined to the cytoplasm, where the virus replicates. When the OTU domain was prevented from entering the nucleus by expressing it as part of the N-terminal 205 kDa of the viral L protein, it continued to block type I interferon signalling, but no longer blocked the TNF-alpha-induced activation of NF-kappaB.

Abstract

We have created a completely helper cell-dependent morbillivirus by modifying the genome to remove the coding sequence of the phosphoprotein (P) and recovering the recombinant virus in a cell line constitutively expressing the P protein. The P protein-deleted virus (P?) grew very inefficiently unless both of the viral accessory proteins (V and C) were also expressed. Growth of the virus was restricted to the P-expressing cell line. The P? virus grew more slowly than the parental virus and expressed much less viral protein in infected cells. The technique could be used to create virus-like particles for use as a vaccine or as antigen in immunological or serological assays.
Barry G, Alberdi P, Schnettler E, Weisheit S, Kohl A, Fazakerley J K, Bell-Sakyi L (2013)

Gene silencing in tick cell lines using small interfering or long double-stranded RNA

Experimental and Applied Acarology 59 (3), 319-338

Abstract

Gene silencing by RNA interference (RNAi) is an important research tool in many areas of biology. To effectively harness the power of this technique in order to explore tick functional genomics and tick-microorganism interactions, optimised parameters for RNAi-mediated gene silencing in tick cells need to be established. Ten cell lines from four economically important ixodid tick genera (Amblyomma, Hyalomma, Ixodes and Rhipicephalus including the sub-species Boophilus) were used to examine key parameters including small interfering RNA (siRNA), double stranded RNA (dsRNA), transfection reagent and incubation time for silencing virus reporter and endogenous tick genes. Transfection reagents were essential for the uptake of siRNA whereas long dsRNA alone was taken up by most tick cell lines. Significant virus reporter protein knockdown was achieved using either siRNA or dsRNA in all the cell lines tested. Optimum conditions varied according to the cell line. Consistency between replicates and duration of incubation with dsRNA were addressed for two Ixodes scapularis cell lines; IDE8 supported more consistent and effective silencing of the endogenous gene subolesin than ISE6, and highly significant knockdown of the endogenous gene 2I1F6 in IDE8 cells was achieved within 48 h incubation with dsRNA. In summary, this study shows that gene silencing by RNAi in tick cell lines is generally more efficient with dsRNA than with siRNA but results vary between cell lines and optimal parameters need to be determined for each experimental system.

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