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

There were a total of 1637 results for your search.
Beard P M, Stevenson K, Pirie A, Rudge K, Buxton D, Rhind S M, Sinclair M C, Wildblood L A, Jones D G, Sharp J M (2001)

Experimental paratuberculosis in calves following inoculation with a rabbit isolate of Mycobacterium avium subsp paratuberculosis

Journal of Clinical Microbiology 39 (9), 3080-3084


The role of wildlife species in the epidemiology of paratuberculosis has been the subject of increased research efforts following the discovery of natural paratuberculosis in free-living rabbits from farms in east Scotland. This paper describes the experimental inoculation of young calves with an isolate of Mycobacterium avium subsp. paratuberculosis recovered from a free-living rabbit. After a 6-month incubation period, all eight calves inoculated with the rabbit isolate had developed histopathological and/or microbiological evidence of M. avium subsp. paratuberculosis infection. Similar results were obtained from a group of calves infected with a bovine isolate of M. avium subsp. paratuberculosis. The virulence of the rabbit isolate for calves demonstrated in this study suggests that rabbits are capable of passing paratuberculosis to domestic ruminants and that wildlife reservoirs of M. avium subsp. paratuberculosis should therefore be considered when formulating control plans for the disease.

Vorechovsky I, Kralovicova J, Tchilian E, Masterman T, Zhang Z P, Ferry B, Misbah S, Chapel H, Webster D, Hellgren D, Anvret M, Hillert J, Hammarstrom L, Beverley P C (2001)

Does 77 -> G in PTPRC modify autoimmune disorders linked to the major histocompatibility locus?

Nature Genetics 29 (1), 22-23
Publisher’s version:


A 77G allele of the gene encoding CD45, also known as the protein tyrosine phosphatase receptor-type C gene (PTPRC), has been associated with multiple sclerosis (MS). Here we determine allele frequencies in large numbers of MS patients, primary immunodeficiencies linked to the major histocompatability complex (MHC) locus and over 1,000 controls to assess whether aberrant splicing of PTPRC caused by the 77C-->G polymorphism results in increased susceptibility to these diseases. Our results show no difference in the frequency of the 77G allele in patients and controls and thus do not support a causative role for the polymorphism in the development of disorders with a strong autoimmune component in etiology.
Salguero F J, Mekonnen T, Ruiz-Villamor E, Sanchez-Cordon P J, Gomez-Villamandos J C (2001)

Detection of monokines in paraffin-embedded tissues of pigs using polyclonal antibodies

Veterinary Research 32 (6), 601-609


Monokines are glycoproteins, synthesised by macrophages, which exert various effects on the organism. The most important monokines are interleukin (IL)-1 alpha, IL-1 beta, tumor necrosis factor (TNF)-alpha and IL-6. This paper reports on immunohistochemical techniques developed for the detection of IL-alpha, IL-1 beta, IL-6 and TNF-alpha in fixed and paraffin-embedded pig tissues (spleen, lymph nodes, thymus, liver and kidney). Different fixatives (buffered formalin, acetic formalin, paraformadehyde-lysine-periodate and Bouin solution), and antigen unmasking techniques (permeabilisation with Tween 20, pronase enzymatic digestion and microwave-citrate buffer) were used. We describe different protocols for detection of monokines using polyclonal antibodies against the studied monokines. No signal was obtained with monoclonal antibodies against pig-TNF-alpha and human IL-1 alpha. Bouin solution was shown to be the best fixative for immunohistochemical detection of IL-1 alpha, TNF-alpha, and IL-6, using permeabilisation with Tween 20 as an unmasking antigen method. Acetic formalin was shown to be the best fixative for IL-1 beta detection, not needing antigen retrieval techniques. Macrophages were identified as the main cytokine-producing cells, although other types of cells also stained positively to some cytokines. These techniques represent valuable tools for studies of the pathogenesis of viral and bacterial diseases, and of the immune system of the pigs.
Fernandez-Fernandez M R, Lucini C, Lopez-Moya J J, Guo H, Martinez-Torrecuadrada J, Casal J I, Mourino M, Plana-Duran J, Rivera J, Rodriguez J F, Montoya M, Del Val M, Garcia J A (2001)

Use of plum pox potyvirus as an expression vector in plants

Molecular Farming. Proceedings of the OECD Workshop held in La Grande Motte, France, 3-6 September, 2000. , 161-172
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Plum pox potyvirus (PPV) is the causal agent of a devastating disease that affects stone fruit trees of the Prunus genus. However, it is also able to infect herbaceous hosts from several different families. This broad host range confers PPV an added value regarding its interest as a putative expression vector in plants. Two strategies have been used to express foreign sequences of interest using vectors based on PPV. The N-terminal region of the capsid protein (CP) has been used to express epitopes on the surface of virion particles. Four different insertion sites were evaluated, but only three of them have rendered viable chimeras. These vectors have shown notable differences in tolerance to sequence insertions. Foreign antigenic peptides expressed in them were easily recognized by specific antibodies. Moreover, sequences cloned at one of these vectors (position 12 of CP) were able to elicit an efficient B-cell response in experimental hosts. Epitope mapping by pepscan analysis and fine tuning of cloning positions are being conducted in an attempt to select optimal sites for epitope insertion. The second approach that we have followed is the use of PPV as an expression vector of whole independent proteins. Two insertion sites have been selected, one between P1 and HC proteins (PPV-XS) and the other between NIb and CP proteins (PPV-NK). A double expression vector has been constructed, which allows production of two foreign proteins with a single vector. ifferent reporter genes have been cloned in both insertion sites. The VP60 structural protein of rabbit hemorrhagic disease virus (RHDV) was also successfully expressed making use of PPV-NK vector. Extracts from the VP60-expressing plants were able to induce a remarkable immune response against RHDV in its natural hosts, rabbits. Moreover, these animals were protected against a lethal challenge with RHDV.
Wurm T, Chen H, Hodgson T, Britton P, Brooks G, Hiscox J A (2001)

Localization to the nucleolus is a common feature of coronavirus nucleoproteins, and the protein may disrupt host cell division

Journal of Virology 75, 9345-9356


The subcellular localization of transmissible gastroenteritis virus (TGEV) and mouse hepatitis virus (MHV) (group I and group II coronaviruses, respectively) nucleoproteins (N proteins) were examined by confocal microscopy. The proteins were shown to localize either to the cytoplasm alone or to the cytoplasm and a structure in the nucleus. This feature was confirmed to be the nucleolus by using specific antibodies to nucleolin, a major component of the nucleolus, and by confocal microscopy to image sections through a cell expressing N protein. These findings are consistent with our previous report for infectious bronchitis virus (group III coronavirus) (J. A. Hiscox et al., J. Virol. 75:506-512, 2001), indicating that nucleolar localization of the N protein is a common feature of the coronavirus family and is possibly of functional significance. Nucleolar localization signals were identified in the domain III region of the N protein from all three coronavirus groups, and this suggested that transport of N protein to the nucleus might be an active process. In addition, our results suggest that the N protein might function to disrupt cell division. Thus, we observed that approximately 30%. of cells transfected with the N protein appeared to be undergoing cell division. The most likely explanation for this is that the N protein induced a cell cycle delay or arrest, most likely in the G(2)/M phase. In a fraction of transfected cells expressing coronavirus N proteins, we observed multinucleate cells and dividing cells with nucleoli (which are only present during interphase). These findings are consistent with the possible inhibition of cytokinesis in these cells.
Carrasco L, Lima J S, Halfen D C, Salguero F J, Sanchez-Cordon P, Becker G (2001)

Systemic aspergillosis in an oiled Magallanic penguin (Spheniscus magellanicus)

Journal of Veterinary Medicine Series B-Infectious Diseases and Veterinary Public Health 48 (7), 551-554


This report describes a case of fatal aspergillosis caused by A. fumigatus during the recovery of an oiled Magallanic penguin. The possible role of aspergillosis as a possible complication responsible for the mortality of penguins surviving the first days of treatment for oil is emphasized.
Seago J E, Chernukhin I V, Newbury S F (2001)

The Drosophila gene twister, an orthologue of the yeast helicase SKI2, is differentially expressed during development

Mechanisms of Development 106 (1-2), 137-141


We have identified and characterized a Drosophila orthologue of SK12, which, in Saccharomyces cerevisiae. is one of the key components in the cytoplasmic 3 ' -5 ' decay of mRNA. The Drosophila orthologue (twister, tst), is expressed as two transcripts which differ in the lengths of their 3 ' -UTRs, with the smaller transcript being particularly abundant in 0-2 h embryos and the larger transcript reaching its highest levels in 6-8 h embryos. TST protein is expressed in two forms which are differentially expressed in adult tissues and throughout development. Differential expression of TST may modulate activity of the mRNA turnover pathway and could have a major impact on the expression of target RNAs.
Gomez-Villamandos J C, Ruiz-Villamor E, Bautista M J, Sanchez C P, Sanchez-Cordon P J, Salguero F J, Jover A (2001)

Morphological and immunohistochemical changes in splenic macrophages of pigs infected with classical swine fever

Journal of Comparative Pathology 125 (2-3), 98-109


Classical swine fever (CSF) was induced in 20 pigs by inoculation with a virulent strain of CSF virus to determine sequential changes (2, 4, 7, 10 and 14 days post-inoculation) in the number and morphology of splenic macrophages (red pulp and lymphoid marginal zone) and thus to assess the role of these cells in the pathogenesis of the disease. The first splenic cells to be infected with CSF virus were macrophages in the marginal zone followed by other macrophage populations. The initial phase of CSF was associated with an increase in splenic macrophage numbers in the marginal zone and a decrease in the red pulp. Subsequently, the numbers in the red pulp increased. The study suggested that infection, mobilization and apoptosis of splenic macrophages play an important role in the spread of CSF virus in vivo. Moreover, the secretory changes that occurred in macrophages in the initial phase of the infection suggested that macrophages release chemical mediators capable of modulating pathogenesis, (C) 2001 Harcourt Publishers Ltd.
Wyde P R, Guzman E, Gilbert B E, Couch R B (2001)

Immunogenicity and protection in mice given inactivated influenza vaccine, MPL, QS-21 or QS-7

4th World Congress on Options for the Control of Influenza, Crete, Greece. 1219, 999-1005


Background: Monophosphoryl lipid A (MPL), QS-21 and QS-7 were evaluated in mice for their ability to increase the immunogenicity and protective efficacy of formalin-inactivated (FI) influenza A/Texas/91 virus vaccine. Freund's incomplete adjuvant (FIA) was used as a positive control. Methods: Mice were inoculated twice, 28 days apart, either intramuscularly (I.M.) with vaccine mixed with phosphate buffered saline, FIA, MPL or QS21, or intranasally J.N.) with vaccine containing QS-21 or QS-7. The mice were bled on days 0, 28 and 49 and challenged I.N. on this last day with live virus. Four days later, the lungs from each animal were assessed for influenza virus. All sera were tested for virus-specific neutralizing (Nt), hemagglutination inhibiting (HI) and ELISA antibodies. Studies to account for the mechanism(s) of adjuvant activity have been initiated. Results: FIA, MPL and QS-21 all enhanced the production of virus-specific antibodies and increased protection from pulmonary virus infection following I.M. administration. Maximal adjuvanticity occurred in groups inoculated with "low" doses of vaccine and in groups administered vaccine mixed with QS-21. Both QS adjuvants exhibited significant adjuvant activity following IN inoculation. Protection correlated best with levels of virus-specific serum Nt and HI antibodies. Conclusions: The present studies support continued development of adjuvants for inactivated influenza virus vaccines.
Tchilian E Z, Wallace D L, Dawes R, Imami N, Burton C, Gotch F, Beverley P C L (2001)

A point mutation in CD45 may be associated with an increased risk of HIV-1 infection

AIDS 15 (14), 1892-1894


The CD45 antigen is essential for normal antigen receptor-mediated signalling in lymphocytes, and different patterns of splicing of CD45 are associated with distinct functions in lymphocytes. Here we show that abnormal CD45 splicing caused by a C77G transversion in exon A of the gene encoding CD45 (PTPRC) is associated with increased susceptibility to HIV-1 infection.


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