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

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Sweeney T R, Moser A H, Shigenaga J K, Grunfeld C, Feingold K R (2006)

Decreased nuclear hormone receptor expression in the livers of mice in late pregnancy

American Journal of Physiology, Endocrinology and Metabolism 290 (6), E1313-E1320
Publisher’s version: https://doi.org/10.1152/ajpendo.00071.2005

Abstract

During the third trimester of pregnancy, there is an increase in serum triglyceride and cholesterol levels. The mechanisms accounting for these changes in lipid metabolism during pregnancy are unknown. We hypothesized that, during pregnancy, the expression of nuclear hormone receptors involved in regulating lipid metabolism would decrease. In 19-day pregnant mice, serum triglyceride and non-HDL cholesterol levels were significantly increased, whereas total cholesterol was slightly decreased, because of a decrease in the HDL fraction. Peroxisome proliferator-activated receptor (PPAR)α, PPARβ/δ, and PPARγ, liver X receptor (LXR)α and LXRβ, farnesoid X receptor (FXR), and retinoid X receptor (RXR)α, RXRβ, and RXRγ mRNA levels were significantly decreased in the livers of 19-day pregnant mice. Additionally, the expressions of thyroid receptor (TR)α, pregnane X receptor, sterol regulatory element-binding proteins (SREBP)-1a, SREBP-1c, SREBP-2, and liver receptor homolog 1 were also decreased, whereas the expression of TRβ, constitutive androstane receptor, and hepatic nuclear factor 4 showed no significant change. mRNA levels of the PPAR target genes carnitine-palmitoyl transferase 1α and acyl-CoA oxidase, the LXR target genes SREBP1c, ATP-binding cassettes G5 and G8, the FXR target gene SHP, and the TR target genes malic enzyme and Spot14 were all significantly decreased. Finally, the expressions of PPARγ coactivator (PGC)-1α and PGC-1β, known activators of a number of nuclear hormone receptors, were also significantly decreased. The decreases in expression of RXRs, PPARs, LXRs, FXR, TRs, SREBPs, and PGC-1s could contribute to the alterations in lipid metabolism during late pregnancy.

Ferris N P, King D P, Reid S M, Shaw A E, Hutchings G H (2006)

Comparisons of original laboratory results and retrospective analysis by real-time reverse transcriptase-PCR of virological samples collected from confirmed cases of foot-and-mouth in the UK in 2001

Veterinary Record 159 (12), 373-378
Publisher’s version: https://veterinaryrecord.bmj.com/content/159/12/373.long

Abstract

There were 2030 designated cases of foot-and-mouth disease (FMD) during the course of the epidemic in the UK in 2001 (including four from Northern Ireland). Samples from 1720 of the infected premises (IPs) were received in the laboratory and examined for either the presence of FMD virus (virological samples from 1421 IPs) or both FMD virus and antibody (virological and serological samples from 255 IPs) or antibody alone (from 44 IPs). The time taken to issue final diagnostic results ranged from a few hours in cases in which positive results were obtained by ELISA on epithelia containing sufficient virus to be detected, to several days for samples containing small amounts of virus requiring amplification through cell culture, negative samples or samples tested for antibody. Two subsets of samples were analysed retrospectively by real-time reverse transcriptase-PCR (RT-PCR); first, epithelia that were negative by both ELISA and virus isolation (VI) in cell culture, and secondly, samples that were negative by ELISA on epithelial suspension but positive by VI. There was broad agreement between the RT-PCR and VI/ELISA combined, except that the RT-PCR procedure did not detect a group of related virus isolates from Wales. These viruses had evidently evolved during the epidemic and had a nucleotide substitution in the RT-PCR probe site, which prevented them from being detected by the routine diagnostic probe. No evidence of FMD virus, antibody or nucleic acid was found in approximately 23 per cent (390 of 1730) of IPs from which samples were received, suggesting that the incidence of FMD during the outbreak may have been over-reported.

Beard P M, Froggatt G C, Smith G L (2006)

Vaccinia virus kelch protein A55 is a 64 kDa intracellular factor that affects virus-induced cytopathic effect and the outcome of infection in a murine intradermal model

Journal of General Virology 87, 1521-1529
Publisher’s version: http://dx.doi.org/10.1099/vir.0.81854-0

Abstract

The vaccinia virus (VACV) protein A55 is a BTB/kelch protein with a broad-complex, tramtrack and bric-a-brac (BTB) domain in the N-terminal region and five kelch repeats in the C-terminal half. The BTB/kelch subgroup of the kelch superfamily of proteins has been associated with a wide variety of functions including regulation of the cytoskeleton. VACV contains three genes predicted to encode BTB/kelch proteins: A55R, F3L and C2L. The A55R gene product has been identified as an intracellular protein of 64 kDa that is expressed late in infection. A VACV strain lacking 93.6% of the A55R open reading frame (v Delta A55) was constructed and found to have an unaltered growth rate in vivo but a different plaque morphology and cytopathic effect, as well as reduced development of VACV-induced Ca2+-independent cell/extracellular matrix adhesion. In a murine intradermal model of VACV infection, a virus lacking the A55R gene induced larger lesions than wild-type and revertant control viruses.

Basto A P, Nix R J, Boinas F, Mendes S, Silva M J, Cartaxeiro C, Portugal R S, Leitao A, Dixon L K, Martins C (2006)

Kinetics of African swine fever virus infection in Ornithodoros erraticus ticks

Journal of General Virology 87 (7), 1863-1871
Publisher’s version: http://dx.doi.org/10.1099/vir.0.81765-0

Abstract

The kinetics of African swine fever virus (ASFV) infection in Ornithodoros erraticus ticks were investigated in specimens collected in the field at different times following an outbreak of the disease in Portugal in 1999 and in ticks infected experimentally with a virus isolated from a tick collected during this outbreak. In ticks collected from the field, initial screening for ASFV was carried out by PCR, followed by attempts to isolate the virus in macrophage cultures. Considering total numbers of ticks tested independently of developmental stages, ASFV DNA was detected in 42.3, 26.4 and 22.4?% of specimens collected at weeks 0, 32 and 63 following the outbreak, respectively. Although virus was not isolated from most of these ticks, the proportion of isolations from large nymphs and adults increased between weeks 0 and 32 from 2 to 9?% and from 5 to 11.5?%, respectively. These results, together with the higher virus titres at week 32, suggest that virus replication occurred. In contrast, virus isolations from small nymphs decreased over this period, from 5 to 1.3?%. At week 63, infection rates decreased for all stages. Experimental infections showed the occurrence of virus replication within 4?weeks post-feeding and maintenance of high titres in almost 100?% of ticks until 20?weeks post-infection. At weeks 41 and 61, a drop in virus titres and infection rates was observed. Relevant to the understanding of African swine fever epidemiology, our results show that ASFV replicates and persists in O. erraticus, but a viral clearance occurs at later times in both natural and experimental infections.
Brocchi E, Bergmann I E, Dekker A, Paton D J, Sammin D J, Greiner M, Grazioli S, De Simone F, Yadin H, Haas B, Bulut N, Malirat V, Neitzert E, Goris N, Parida S, Sorensen K, De Clercq K (2006)

Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus

Vaccine 24, 6966-6979
Publisher’s version: http://dx.doi.org/10.1016/j.vaccine.2006.04.050

Abstract

To validate the use of serology in substantiating freedom from infection after foot-and-mouth disease (FMD) outbreaks have been controlled by measures that include vaccination, 3551 sera were tested with six assays that detect antibodies to the non-structural proteins of FMD virus. The sera came from naïve, vaccinated, infected and vaccinated-and-infected animals; two-thirds from cattle, the remainder from sheep and pigs. The assays were covariant for sensitivity, but not necessarily for specificity. A commercial kit from Cedi-diagnostics and an in-house assay from IZS-Brescia were comparable to the NCPanaftosa-screening index method described in the Diagnostic Manual of the World Animal Health Organisation. Using these three tests the specificity and sensitivity for the detection of carriers in vaccinated cattle approaches or exceeds 99% and 90%, respectively.
Cox S J, Voyce C, Parida S, Reid S M, Hamblin P A, Hutchings G, Paton D J, Barnett P V (2006)

Effect of emergency FMD vaccine antigen payload on protection, sub-clinical infection and persistence following direct contact challenge of cattle

Vaccine 24, 3184-3190
Publisher’s version: http://dx.doi.org/10.1016/j.vaccine.2006.01.037

Abstract

Previous work, in sheep vaccinated with emergency foot-and-mouth disease (FMD) vaccine, indicated the benefit of increasing the antigen payload in inhibiting local virus replication and consequently persistence following an indirect aerosol challenge with a virus homologous to the vaccine strain. The work presented here investigates this possibility further using cattle and a more severe semi-heterologous direct contact challenge. The quantitative dynamics of virus replication and excretion in both vaccinated and non-vaccinated cattle following challenge are examined. Two experiments were carried out each involving 20 vaccinated and 5 non-vaccinated cattle. An O1 Manisa vaccine (18 PD50) was used for the first, previously reported experiment [Cox SJ, Voyce C, Parida S, Reid SM, Hamblin PA, Paton DJ, et al. Protection against direct contact challenge following emergency FMD vaccination of cattle and the effect on virus excretion from the oropharynx. Vaccine 2005;23:1106–13]. The same vaccine was used for the second experiment described in this paper except the antigen payload was increased 10-fold per bovine dose, resulting in significantly higher FMD virus neutralising antibody titres prior to challenge. Twenty-one days post-vaccination the cattle received a 5-day direct contact challenge with FMD virus from five further non-vaccinated cattle infected 24 h earlier with O UKG 34/2001. All vaccinated cattle regardless of antigen payload were protected against clinical disease. Sub-clinical oropharyngeal infection was detected in animals from both experiments but the level of virus replication shortly after direct contact challenge was significantly reduced in vaccinated animals. Cattle immunised with the 10-fold antigen payload cleared the virus more readily and consequently at 28 days post-challenge fewer animals were persistently infected compared to the single strength vaccine. Following a severe challenge, the results from both experiments show that use of emergency vaccine can prevent or decrease local virus replication and thereby dramatically reduce the amount of virus released into the environment, particularly during the early post-exposure period. Additionally, increasing the antigen payload of the vaccine may reduce sub-clinical infection, leading to fewer persistently infected virus carrier animals.
Dafa'alla T H, Condon G C, Condon K C, Phillips C E, Morrison N I, Jin L, Epton M J, Fu G, Alphey L (2006)

Transposon-free insertions for insect genetic engineering

Nature Biotechnology 24 (7), 820-821
Publisher’s version: http://dx.doi.org/10.1038/nbt1221

Abstract

Methods involving the release of transgenic insects in the field hold great promise for controlling vector-borne diseases and agricultural pests. Insect transformation depends on nonautonomous transposable elements as gene vectors. The resulting insertions are stable in the absence of suitable transposase, however, such absence cannot always be guaranteed. We describe a method for post-integration elimination of all transposon sequences in the pest insect Medfly, Ceratitis capitata. The resulting insertions lack transposon sequences and are therefore impervious to transposase activity.

Dawes R, Hennig B, Irving W, Petrova S, Boxall S, Ward V, Wallace D, Macallan D C, Thursz M, Hill A, Bodmer W, Beverley P C L, Tchilian E Z (2006)

Altered CD45 expression in C77G carriers influences immune function and outcome of hepatitis C infection

Journal of Medical Genetics 43 (8), 678-684
Publisher’s version: http://dx.doi.org/10.1136/jmg.2005.040485

Abstract

Background: A polymorphism in exon 4 (C77G) of CD45 that alters CD45 splicing has been associated with autoimmune and infectious diseases in humans. Objective: To investigate the effect of C77G in hepatitis C virus (HCV) infected individuals and study the phenotype and function of peripheral blood mononuclear cells (PBMC) from healthy and hepatitis C infected C77G carriers. Results: C77G individuals showed an increased proportion of primed CD45RA and effector memory CD8 T cells and more rapid activation of the lymphocyte specific protein tyrosine kinase (Lck) following CD3 stimulation. Transgenic mice with CD45 expression mimicking that in human C77G variants had more activated/memory T cells, more rapid proliferative responses, and activation of Lck. Conclusions: Changes in CD45 isoform expression can alter immune function in human C77G variants and CD45 transgenic mice. The C77G allele may influence the outcome of HCV infection.
Dawes R, Petrova S, Liu Z, Wraith D, Beverley P C L, Tchilian E Z (2006)

Combinations of CD45 isoforms are crucial for immune function and disease

Journal of Immunology 176 (6), 3417-3425
Publisher’s version: http://www.jimmunol.org/content/176/6/3417.abstract

Abstract

Expression of the CD45 Ag in hemopoietic cells is essential for normal development and function of lymphocytes, and both mice and humans lacking expression exhibit SCID. Human genetic variants of CD45, the exon 4 C77G and exon 6 A138G alleles, which after the pattern of CD45 isoform expression, are associated with autoimmune and infectious diseases. We constructed transgenic mice expressing either an altered level or combination of CD45 isoforms. We show that the total level of CD45 expressed is crucial for normal TCR signaling, lymphocyte proliferation, and cytokine production. Most importantly, transgenic lines with a normal level, but altered combinations of CD45 isoforms, CD45(RABC/+) and CD45(RO/+) mice, which mimic variant CD45 expression in C77G and A138G humans, show more rapid onset and increased severity of experimental autoimmune encephalomyelitis. CD45(RO/+) cells produce more TNF-alpha and IFN-gamma. Thus, for the first time, we have shown experimentally that it is the combination of CD45 isoforms that affects immune function and disease.
Frolich K, Hamblin C, Parida S, Tuppurainen E, Schettler E (2006)

Serological survey for potential disease agents of free-ranging cervids in six selected national parks from Germany

Journal of Wildlife Diseases 42, 836-843
Publisher’s version: http://dx.doi.org/10.7589/0090-3558-42.4.836

Abstract

A total of 164 blood samples, collected from free-ranging red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama danw) in six German national parks (NP) between 2000 and 2002, were assayed for antibodies against nine viral disease agents. Antibodies were only detected against the a-herpesviruses; specifically, bovine herpesvirus-1 (BHV-1) (22 of 157,14%), cervid herpesvirus-1 (17 of 157, 10.8%), and caprine herpesvirus-1 (11 of 159, 6.9%). Titers ranged from 4 to 102. Most of the seropositive sera, and those with the highest antibody titers, were from red and roe deer in the Harz and Hochharz NP, which are connected and allow migration between the two. The distribution and specificity of antibodies detected in individual deer suggests that the three alpha-herpesviruses are circulating in these deer populations. No antibodies were detected against bovine viral diarrhea virus, epizootic hemorrhagic disease virus, bovine leukemia virus, bluetongue virus, foot-and-mouth disease virus, or sheep and goat poxvirus.

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