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

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Faburay B, Munstermann S, Geysen D, Bell-Sakyi L, Ceesay A, Bodaan C, Jongejan F (2005)

Point seroprevalence survey of Ehrlichia ruminantium infection in small ruminants in The Gambia

Clinical and Diagnostic Laboratory Immunology 12 (4), 508-512


Using the MAP1-B enzyme-linked immunosorbent assay, we tested 1,318 serum samples collected from sheep and goats at 28 sites in the five divisions of The Gambia to determine the Ehrlichia ruminantium seroprevalence rates and to assess the risk for heartwater. About half (51.6 %) of 639 sheep were positive, with seroprevalence rates per site varying between 6.9 % and 100 %. The highest seroprevallence was detected in the western part of the country (88.1 % in the Western Division and 62.1 % in the Lower River Division). Sheep in the two easterly divisions (Central River and Upper River divisions) showed the lowest seroprevalence of 29.3 % and 32.4 %, respectively, while those in the North Bank Division showed an intermediate prevalence of 40.6 %. In goats, less than one-third (30.3 %) of 679 animals tested were positive. The highest seroprevalence was detected in goats in the North Bank Division (59 %) and Western Division (44.1 %). Goats in the Lower River Division showed an intermediate level of 21.9 %, whereas the lowest rates were found in the eastern part of the country (4.8 % in the Central River Division and 2.3 % in the Upper River Division). At nearly all sites, seroprevalence rates were higher in sheep than in goats. The results show a gradient of increasing heartwater risk for susceptible small ruminants from the east to the west of The Gambia. These findings need to be taken into consideration when future livestock-upgrading programs are implemented.
Carpentier G S, Fleury M J J, Touze A, Sadeyen J M, Tourne S, Sizaret P Y, Coursaget P (2005)

Mutations on the FG surface loop of human papillomavirus type 16 major capsid protein affect recognition by both type-specific neutralizing antibodies and cross-reactive antibodies

Journal of Medical Virology 77 (4), 558-565


The aim of this study was to further characterize the conformational neutralizing epitopes present on the surface-exposed FG loop of human papillomavirus (HPV) type 16 L1 major capsid protein. We have generated previously two chimeric L1 proteins by insertion of a foreign peptide encoding an epitope of the hepatitis B core (HBc) antigen within the FG loop. In addition, three other chimeric L1 proteins were obtained by replacing three different FG loop sequences by the HBc motif and three others by point mutations. All these chimeric L1 proteins retained the ability to self-assemble into virus-like particles (VLPs), with the exception of the mutant with substitution of the L1 sequence 274279 by the HBc motif. The eight chimeric VLPs were then analyzed for differential reactivity wit a set of six HPV-16 and HPV-31 monoclonal antibodies that bound to conformational and linear epitopes. The binding patterns of these monoclonal antibodies confirmed that the FG loop contained or contributed to neutralizing conformational epitopes. The results obtained suggested that the H31.F7 antibody, an anti-HPV-31 cross-reacting and neutralizing antibody, recognized a conformational epitope situated before the 266-271 sequence. In addition, H16.E70 neutralizing antibody reactivity was reduced with L1 VLPs with an Asn to Ala point mutation at position 270, suggesting that Asn is a part of the epitope recognized by this antibody. This study contributes to the understanding of the antigenic structure of HPV-16 and -31 L1 proteins by confirming that the FG loop contributes to neutralizing epitopes and suggesting the existence of both type-specific and cross-reactive conformational epitopes within the FG loop.
Brackenbury L S, Carr B V, Stamataki Z, Prentice H, Lefevre E A, Howard C J, Charleston B (2005)

Identification of a cell population that produces alpha/beta interferon in vitro and in vivo in response to noncytopathic bovine viral diarrhea virus

Journal of Virology 79, 7738-7744


In vitro infection of bovine cells of many origins with the cytopathogenic bovine viral diarrhea virus (cpBVDV) results in the induction of alpha/beta interferon (IFN-alpha/beta), whereas noncytopathogenic BVDV (ncpBVDV) isolates have been shown not to induce IFN-alpha/beta in vitro. Similarly, epBVDV induces IFN-alpha/beta in the early bovine fetus, but ncpBVDV does not. However, acute infection of naive cattle with ncpBVDV results in IFN-alpha/beta production. In this study, we identified and characterized a minor population of cells, present in lymph nodes that produce IFN-alpha in response to ncpBVDV. These cells expressed the myeloid markers CD14, CD11b, and CD172a but did not express CD4 and CD45RB. We also established that these cells produced IFN-alpha in the absence of detectable productive infection.
Britton P, Evans S, Dove B, Davies M, Casais R, Cavanagh D (2005)

Generation of a recombinant avian coronavirus infectious bronchitis virus using transient dominant selection

Journal of Virological Methods 123, 203-211


A reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) has been described in which a full-length cDNA, corresponding to the IBV (Beaudette-CK) genome, was inserted into the vaccinia virus genome following in vitro assembly of three contiguous cDNAs [Casais, R., Thiel. V.. Siddell, S.G., Cavanagh, D., Britton, P., 2001. Reverse genetics system for the avian coronavirus infectious bronchitis virus. J. Virol. 75, 12359-12369]. The method has subsequently been used to generate a recombinant IBV expressing a chimaeric S gene [Casais, R., Dove, B., Cavanagh, D., Britton, P., 2003. Recombinant avian infectious bronchitis virus expressing a heterologous spike gene demonstrates that the spike protein is a determinant of cell tropism. J. Virol. 77, 9084-9089]. Use of vaccinia virus as a vector for the full-length cDNA of the IBV genome has the advantage that modifications can be made to the IBV cDNA using homologous recombination, a method frequently used to insert and delete sequences from the vaccinia virus genome. We describe the use of homologous recombination as a method for modifying the Beaudette full-length cDNA, within the vaccinia virus genome, without the requirement for in vitro assembly of the IBV cDNA. To demonstrate the feasibility of the method we exchanged the ectodomain of the Beaudette spike gene for the corresponding region from IBV M41 and generated two recombinant infectious bronchitis viruses (rIBVs) expressing the chimaeric S protein, validating the method as an alternative way for generating rIBVs.
Edwards M J, Buchatska E, Ashton M, Montoya M, Bickle Q D, Borrow P (2005)

Reciprocal immunomodulation in a schistosome and hepatotropic virus coinfection model

Journal of Immunology 175 (10), 6275-6285


Human coinfection with the helminth parasite Schistosoma mansoni and hepatitis B and hepatitis C viruses is associated with increased hepatic viral burdens and severe liver pathology. In this study we developed a murine S. mansonillymphocytic choriomeningitis virus (LCMV) coinfection model that reproduces the enhanced viral replication and liver pathology observed in human coinfections, and used this model to explore the mechanisms involved. Viral coinfection during the Th2-dominated granulomatous phase of the schistosome infection resulted in induction of a strong LCMV-specific T cell response, with infiltration of high numbers of LCMV-specific IFN-gamma-producing CD8(+) cells into the liver. This was associated with suppression of production of the Th2 cytokines dominant during S. mansoni infection and a rapid increase in morbidity, linked to hepatotoxicity. Interestingly, the liver of coinfected mice was extremely susceptible to viral replication. This correlated with a reduced intrahepatic type I IFN response following virus infection. Schistosome egg Ags were found to suppress the type I IFN response induced in murine bone marrow-derived dendritic cells by polyinosinic-polycytidylic acid. These results suggest that suppression of the antiviral type I IFN response by schistosome egg Ags in vivo predisposes the liver to enhanced viral replication with ensuing immunopathological consequences, findings that maybe paralleled inhuman schistosome/hepatotropic virus coinfections.
Beard P M, Baines J D (2004)

The DNA cleavage and packaging protein encoded by the U(L)33 gene of herpes simplex virus 1 associates with capsids

Virology 324 (2), 475-482


The U(L)33 gene of herpes simplex virus 1 (HSV-1) encodes a protein (pU(L)33) that is essential for the cleavage and packaging of concatameric herpesvirus DNA into preformed capsids. Previous data have suggested that the U(L)33 protein interacts with the cleavage and packaging proteins encoded by U(L)15 and U(L)28 that are known to associate with capsids. Examination of purified A capsids that lack DNA and are derived from aborted packaging events, B capsids that lack DNA, and C capsids that contain DNA revealed an association of the U(L)33 protein with all three capsid types. More U(L)33 protein was detected in A capsids than was present in B capsids. Capsid association was susceptible to guanidine-HCl treatment and independent of the presence of U(L)15 or U(L)28. Capsid association of pU(L)33 was also independent of U(L)6, which is believed to encode the portal into which DNA is inserted. These data suggest that pU(L)33 may act as part of the capsid-associated molecular machinery that translocates cleaved genomic DNA into the capsid interior.

Beard P M, Duffy C, Baines J D (2004)

Quantification of the DNA cleavage and packaging proteins U(L)15 and U(L)28 in A and B capsids of herpes simplex virus type 1

Journal of Virology 78 (3), 1367-1374


The proteins produced by the herpes simplex virus type 1 (HSV-1) genes U(L)15 and U(L)28 are believed to form part of the terminase enzyme, a protein complex essential for the cleavage of newly synthesized, concatameric herpesvirus DNA and the packaging of the resultant genome lengths into preformed capsids. This work describes the purification of recombinant forms of PU(L)15 and pU(L)28, which allowed the calculation of the average number of copies of each protein in A and B capsids and in capsids lacking the putative portal encoded by U(L)6. On average, 1.0 (+/-0.29 [standard deviation]) copies of PU(L)15 and 2.4 (+/-0.97) copies of pU(L)28 were present in B capsids, 1.2 (+/-0.72) copies of PU(L)15 and 1.5 (+/-0.86) copies of pU(L)28 were found in mutant capsids lacking the putative portal protein pU(L)6, and approximately 12.0 (+/-5.63) Copies of pU(L)15 and 0.6 (+/-0.32) copies of pU(L)28 were present in each A capsid. These results suggest that the packaging machine is partly comprised of approximately 12 copies of pU(L)15, as found in A capsids, with wild-type B and mutant U(L)6(-) capsids containing an incomplete complement of cleavage and packaging proteins. These results are consistent with observations that B capsids form by default in the absence of packaging machinery in vitro and in vivo. In contrast, A capsids may be the result of initiated but aborted attempts at DNA packaging, resulting in the retention of at least part of the DNA packaging machinery.

Hodgson T, Casais R, Dove B, Britton P, Cavanagh D (2004)

Recombinant infectious bronchitis coronavirus Beaudette with the spike protein gene of the pathogenic M41 strain remains attenuated but induces protective immunity

Journal of Virology 78, 13804-13811


We have replaced the ectodomain of the spike (S) protein of the Beaudette strain (Beau-R; apathogenic for Gallus domesticus chickens) of avian infectious bronchitis coronavirus (IBV) with that from the pathogenic M41 strain to produce recombinant IBV BeauR-M41(S). We have previously shown that this changed the tropism of the virus in vitro (R. Casais, B. Dove, D. Cavanagh, and P. Britton, J. Virol. 77:9084-9089, 2003). Herein we have assessed the pathogenicity and immunogenicity of BeauR-M41(S). There were no consistent differences in pathogenicity between the recombinant BeauR-M41(S) and its apathogenic parent Beau-R (based on snicking, nasal discharge, wheezing, watery eyes, rales, and ciliostasis in trachea), and both replicated poorly in trachea and nose compared to M41; the S protein from the pathogenic M41 had not altered the apathogenic nature of Beau-R. Both Beau-R and BeauR-M41(S) induced protection against challenge with M41 as assessed by absence of recovery of challenge virus and nasal exudate. With regard to snicking and ciliostasis, BeauR-M41(S) induced greater protection (seven out of nine chicks [77%]; assessed by ciliostasis) than Beau-R (one out of nine; 11%) but less than M41 (100%). The greater protection induced by BeauR-M41(S) against M41 may be related to the ectodomain of the spike protein of Beau-R differing from that of M41 by 4.1%; a small number of epitopes on the S protein may play a disproportionate role in the induction of immunity. The results are promising for the prospects of S-gene exchange for IBV vaccine development.
Tchilian E Z, Dawes R, Hyland L, Montoya M, Le Bon A, Borrow P, Hou S, Tough D, Beverley P C L (2004)

Altered CD45 isoform expression affects lymphocyte function in CD45 Tg mice

International Immunology 16 (9), 1323-1332


Transgenic mice have been constructed expressing high (CD45RABC) and low (CD45R0) molecular weight CD45 isoforms on a CD45(-/-) background. Phenotypic analysis and in vivo challenge of these mice with influenza and lymphocytic choriomeningitis viruses shows that T cell differentiation and peripheral T cell function are related to the level of CD45 expression but not to which CD45 isoform is expressed. In contrast, B cell differentiation is not restored, irrespective of the level of expression of a single isoform. All CD45 trangenic mice have T cells with an activated phenotype and increased T cell turnover. These effects are more prominent in CD8 than CD4 cells. The transgenic mice share several properties with humans expressing variant CD45 alleles and provide a model to understand immune function in variant individuals.
Bell-Sakyi L, Koney E B M, Dogbey O, Walker A R (2004)

Ehrlichia ruminantium seroprevalence in domestic ruminants in Ghana - I. Longitudinal survey in the Greater Accra Region

Veterinary Microbiology 100 (3-4), 175-188


Serum samples collected monthly over a 34-month period from cattle, sheep and goats in the Greater Accra Region of Ghana were tested for antibodies to Ehrlichia (previously Cowdria) ruminantium, the causative agent of heartwater, by polyclonal competitive ELISA (PC-ELISA). Maternal antibodies, detected in about half of animals followed from under 1 month old, declined to negative levels within 2-4 months. Amblyomma variegatum tick vectors were present on livestock in rural areas throughout the year, and first seroconversion occurred at any age, although the majority of calves seroconverted between 1 and 10 months old, sheep by 11 months, and goats by 7 months. All the cattle in the study became seropositive by 20 months of age, except one animal which subsequently died of heartwater. Following seroconversion, 25% of bovine sera tested negative in the PC-ELISA. Just over half the sheep in the survey seroconverted before or during the study period; following seroconversion, less than 3% of ovine sera became PC-ELISA negative. About a quarter of the goats seroconverted, and 34% of their post-seroconversion sera tested negative in the PC-ELISA. Overall, the serology indicated that virtually all cattle on the survey farms were exposed to E. ruminantium without suffering disease, but that a substantial proportion of sheep and goats escaped exposure and thus formed a susceptible population. E. ruminantium was detected in brains of 14, 36 and 4% of cattle, sheep and goats submitted for post mortem at the Accra Veterinary Laboratory, indicating that sheep were most at risk from heartwater disease.


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