266 results on '"James P. Stewart"'
Search Results
102. A validated gene expression model of high-risk multiple myeloma is defined by deregulated expression of genes mapping to chromosome 1
- Author
-
Elias Anaissie, Joshua Epstein, Bart Barlogie, David R. Williams, Guido Tricot, Yazan Alsayed, James P. Stewart, Yongsheng Huang, Michele Cottler-Fox, Bob Kordsmeier, Jeffrey R. Sawyer, Ichiro Hanamura, Klaus Hollmig, John D. Shaughnessy, Ronald C. Walker, Fenghuang Zhan, Maurizio Zangari, Hongwei Xu, Frits van Rhee, Simona Colla, Abid Mohiuddin, Yan Xiao, Mauricio Pineda-Roman, Christopher Randolph, Somashekar G. Krishna, Bart Burington, and John Crowley
- Subjects
Multivariate analysis ,Immunology ,Chromosomal translocation ,Biology ,Bioinformatics ,Biochemistry ,Cohort Studies ,Gene mapping ,Recurrence ,Risk Factors ,medicine ,Humans ,Survival rate ,Multiple myeloma ,Aged ,Models, Genetic ,Microarray analysis techniques ,Gene Expression Profiling ,Hazard ratio ,Chromosome Mapping ,Cell Biology ,Hematology ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Survival Rate ,Gene expression profiling ,Chromosomes, Human, Pair 1 ,Multigene Family ,Multiple Myeloma - Abstract
To molecularly define high-risk disease, we performed microarray analysis on tumor cells from 532 newly diagnosed patients with multiple myeloma (MM) treated on 2 separate protocols. Using log-rank tests of expression quartiles, 70 genes, 30% mapping to chromosome 1 (P < .001), were linked to early disease-related death. Importantly, most up-regulated genes mapped to chromosome 1q, and down-regulated genes mapped to chromosome 1p. The ratio of mean expression levels of up-regulated to down-regulated genes defined a high-risk score present in 13% of patients with shorter durations of complete remission, event-free survival, and overall survival (training set: hazard ratio [HR], 5.16; P < .001; test cohort: HR, 4.75; P < .001). The high-risk score also was an independent predictor of outcome endpoints in multivariate analysis (P < .001) that included the International Staging System and high-risk translocations. In a comparison of paired baseline and relapse samples, the high-risk score frequency rose to 76% at relapse and predicted short postrelapse survival (P < .05). Multivariate discriminant analysis revealed that a 17-gene subset could predict outcome as well as the 70-gene model. Our data suggest that altered transcriptional regulation of genes mapping to chromosome 1 may contribute to disease progression, and that expression profiling can be used to identify high-risk disease and guide therapeutic interventions.
- Published
- 2006
103. Regulation and role of REST and REST4 variants in modulation of gene expression in in vivo and in vitro in epilepsy models
- Author
-
Judy M. Coulson, Kate Haddley, Anja Kipar, David J. Hughes, Noel J. Buckley, K E Chandler, James P. Stewart, Mark Howard, Matthew C. Walker, John P. Quinn, Nikolai D. Belyaev, and E M Spencer
- Subjects
Gene isoform ,Male ,Repressor ,Fluorescent Antibody Technique ,Hippocampal formation ,Biology ,Hippocampus ,lcsh:RC321-571 ,Rats, Sprague-Dawley ,Organ Culture Techniques ,Status Epilepticus ,Genes, Reporter ,Seizures ,Gene expression ,NRSF ,Excitatory Amino Acid Agonists ,Gene silencing ,Animals ,heterocyclic compounds ,RNA, Messenger ,Gene ,Transcription factor ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,Cells, Cultured ,Reporter gene ,Kainic Acid ,Microscopy, Confocal ,Epilepsy ,Reverse Transcriptase Polymerase Chain Reaction ,REST ,Neuropeptides ,Preprotachykinin A ,Molecular biology ,Rats ,Repressor Proteins ,Neurology ,Gene Expression Regulation ,Electrophoresis, Polyacrylamide Gel ,Tachykinin ,Transcription Factors - Abstract
Repressor element-1 silencing transcription factor (REST) is a candidate modulator of gene expression during status epilepticus in the rodent. In such models, full-length REST and the truncated REST4 variant are induced and can potentially direct differential gene expression patterns. We have addressed the regulation of these REST variants in rodent hippocampal seizure models and correlated this with expression of the proconvulsant, substance P encoding, PPT-A gene. REST and REST4 were differentially regulated following kainic acid stimulus both in in vitro and in vivo models. REST4 was more tightly regulated than REST in both models and its transient expression correlated with that of the differential regulation of PPT-A. Consistent with this, overexpression of a truncated REST protein (HZ4, lacking the C-terminal repression domain) increased expression of the endogenous PPT-A gene. Similarly the proximal PPT-A promoter reporter gene construct was differentially regulated by the distinct REST isoforms in hippocampal cells with HZ4 being the major inducer of increased reporter expression. Furthermore, REST and REST4 proteins were differentially expressed and compartmentalized within rat hippocampal cells in vitro following noxious stimuli. This differential localization of the REST isoforms was confirmed in the CA1 region following perforant path and kainic acid induction of status epilepticus in vivo. We propose that the interplay between REST and REST4 alter the expression of proconvulsant genes, as exemplified by the PPT-A gene, and may therefore regulate the progression of epileptogenesis.
- Published
- 2006
104. The molecular classification of multiple myeloma
- Author
-
Ronald Walker, Bart Barlogie, Frits van Rhee, Jeffrey R. Sawyer, Sushil K. Gupta, Elias Anaissie, Ichiro Hanamura, Guido J Tricot, John D. Shaughnessy, Joshua Epstein, Klaus Hollmig, Fenghuang Zhan, Maurizio Zangari, Bart Burington, Mauricio Pineda-Roman, Yongsheng Huang, Simona Colla, Shmuel Yaccoby, John Crowley, and James P. Stewart
- Subjects
Syndecans ,Myeloid ,Plasma Cells ,Immunology ,Biology ,Biochemistry ,Cyclin D ,Cyclins ,Gene expression ,medicine ,Cluster Analysis ,Humans ,RNA, Messenger ,RNA, Neoplasm ,Gene ,Multiple myeloma ,Oligonucleotide Array Sequence Analysis ,Membrane Glycoproteins ,Neoplasia ,Gene Expression Profiling ,Chromosome Mapping ,Cell Biology ,Hematology ,Prognosis ,medicine.disease ,Gene expression profiling ,medicine.anatomical_structure ,MAFB ,Data Interpretation, Statistical ,Cancer research ,Proteoglycans ,PAX5 ,Syndecan-1 ,Hyperdiploidy ,Multiple Myeloma - Abstract
To better define the molecular basis of multiple myeloma (MM), we performed unsupervised hierarchic clustering of mRNA expression profiles in CD138-enriched plasma cells from 414 newly diagnosed patients who went on to receive high-dose therapy and tandem stem cell transplants. Seven disease subtypes were validated that were strongly influenced by known genetic lesions, such as c-MAF– and MAFB-, CCND1- and CCND3-, and MMSET-activating translocations and hyperdiploidy. Indicative of the deregulation of common pathways by gene orthologs, common gene signatures were observed in cases with c-MAF and MAFB activation and CCND1 and CCND3 activation, the latter consisting of 2 subgroups, one characterized by expression of the early B-cell markers CD20 and PAX5. A low incidence of focal bone disease distinguished one and increased expression of proliferation-associated genes of another novel subgroup. Comprising varying fractions of each of the other 6 subgroups, the proliferation subgroup dominated at relapse, suggesting that this signature is linked to disease progression. Proliferation and MMSET-spike groups were characterized by significant overexpression of genes mapping to chromosome 1q, and both exhibited a poor prognosis relative to the other groups. A subset of cases with a predominating myeloid gene expression signature, excluded from the profiling analyses, had more favorable baseline characteristics and superior prognosis to those lacking this signature.
- Published
- 2006
105. High-resolution genomic profiles define distinct clinico-pathogenetic subgroups of multiple myeloma patients
- Author
-
Kenneth C. Anderson, Daniel R. Carrasco, Giovanni Tonon, Alexei Protopopov, Deepak B. Khatry, Ronald A. DePinho, Ichiro Hanamura, John D. Shaughnessy, Lynda Chin, Yunyu Zhang, Raktim Sinha, Fenghuang Zhan, Bin Feng, Marina Protopopova, Kumar Sukhdeo, Yongsheng Huang, Owen W. Stephens, James P. Stewart, Cameron Brennan, Bart Barlogie, Carrasco, Dr, Tonon, G, Huang, Y, Zhang, Y, Sinha, R, Feng, B, Stewart, Jp, Zhan, F, Khatry, D, Protopopova, M, Protopopov, A, Sukhdeo, K, Hanamura, I, Stephens, O, Barlogie, B, Anderson, Kc, Chin, L, Shaughnessy, Jr., Jd, and Brennan, C and DePinho RA
- Subjects
Cancer Research ,Gene Dosage ,Genomics ,CELLCYCLE ,Biology ,Gene dosage ,Genome ,Disease-Free Survival ,medicine ,Chromosomes, Human ,Humans ,Multiple myeloma ,Genetics ,Regulation of gene expression ,Genome, Human ,Gene Expression Profiling ,Cell Biology ,medicine.disease ,Prognosis ,Diploidy ,Human genetics ,Gene expression profiling ,Gene Expression Regulation, Neoplastic ,Oncology ,Multiple Myeloma ,Comparative genomic hybridization - Abstract
SummaryTo identify genetic events underlying the genesis and progression of multiple myeloma (MM), we conducted a high-resolution analysis of recurrent copy number alterations (CNAs) and expression profiles in a collection of MM cell lines and outcome-annotated clinical specimens. Attesting to the molecular heterogeneity of MM, unsupervised classification using nonnegative matrix factorization (NMF) designed for array comparative genomic hybridization (aCGH) analysis uncovered distinct genomic subtypes. Additionally, we defined 87 discrete minimal common regions (MCRs) within recurrent and highly focal CNAs. Further integration with expression data generated a refined list of MM gene candidates residing within these MCRs, thereby providing a genomic framework for dissection of disease pathogenesis, improved clinical management, and initiation of targeted drug discovery for specific MM patients.
- Published
- 2006
- Full Text
- View/download PDF
106. Differential Transcription of Ovine Herpesvirus 2 Genes in Lymphocytes from Reservoir and Susceptible Species
- Author
-
George C. Russell, Leenadevi Thonur, James P. Stewart, and David M. Haig
- Subjects
Genes, Viral ,Transcription, Genetic ,DNA polymerase ,T-Lymphocytes ,Gene Expression ,Genome ,Peripheral blood mononuclear cell ,Cell Line ,Gammaherpesvirinae ,Species Specificity ,Viral life cycle ,Transcription (biology) ,Virology ,Gene expression ,Genetics ,Animals ,Molecular Biology ,Gene ,Disease Reservoirs ,Sheep ,Base Sequence ,biology ,General Medicine ,Doxorubicin ,Cell culture ,DNA, Viral ,Azacitidine ,biology.protein ,Nucleic Acid Conformation ,Cattle ,Rabbits - Abstract
Ovine herpesvirus 2 (OvHV-2) is a lymphotropic gammaherpesvirus that asymptomatically infects most sheep, but causes malignant catarrhal fever in cattle, bison, pigs and deer. There is no permissive cell culture system but OvHV-2-infected T lymphocytes can be cultured from diseased animals. We showed that the OvHV-2 genome was in a circular conformation in sheep peripheral blood mononuclear cells and that the latency-associated ORF73 was transcribed, while expression of the productive cycle genes ORF9 (DNA polymerase) and ORF50 (R-transactivator) was barely detectable, suggestive of latency. Doxorubicin treatment of these cells induced the appearance of linear viral DNA and transcription of productive cycle genes along with several viral unique genes. In contrast, cultured T cells from diseased cattle and rabbits contained a mixture of circular and linear genome configurations indicative of a mixture of latently- and productively-infected cells. Most of the OvHV-2 unique genes were transcribed in these cells but ORF50 expression was only seen after doxorubicin treatment indicating a 'leaky' latent pattern of gene expression. 5-azacytidine treatment increased the proportion of circular DNA and inhibited the expression of most of the OvHV-2 unique genes except Ov2.5 (vIL-10) and Ov4.5 (Bcl-2 homologue) in the cattle cell line. These studies provide key insights into the differences in OvHV-2 gene expression in cells from reservoir and susceptible species and, for the first time, an in vitro system for studying the latent and productive phases of the OvHV-2 virus life cycle.
- Published
- 2006
107. T-Cell Responses to the M3 Immune Evasion Protein of Murid Gammaherpesvirus 68 Are Partially Protective and Induced with Lytic Antigen Kinetics
- Author
-
Ondine Silvia, James P. Stewart, Sarah G. Crist, Douglas C. Donovan, Edward J. Usherwood, and Joshua J. Obar
- Subjects
Rhadinovirus ,T-Lymphocytes ,T cell ,Immunology ,Epitopes, T-Lymphocyte ,Viral Plaque Assay ,CD8-Positive T-Lymphocytes ,Biology ,Microbiology ,Virus ,Epitope ,DNA vaccination ,Interferon-gamma ,Mice ,Viral Proteins ,Immune system ,Antigen ,Virology ,Vaccines, DNA ,medicine ,Animals ,Cytotoxic T cell ,Antigens, Viral ,Lung ,Mice, Inbred BALB C ,Viral Vaccines ,Herpesviridae Infections ,respiratory system ,medicine.anatomical_structure ,Lytic cycle ,Insect Science ,Pathogenesis and Immunity ,Epitope Mapping - Abstract
DNA vaccination with theM3gene, encoding an immune evasion molecule expressed during both the acute lytic and persistent phases of murid gammaherpesvirus 68 infection, yielded a significantly lower titer of virus in the lung than controls. The protection seen was dependent on T cells, and we mapped an epitope recognized by CD8 T cells. The immune response to this epitope follows the same kinetics as lytic cycle antigens, despite the fact that this gene is expressed in both lytic and persistent stages of infection. This has important implications for our understanding of T-cell responses to putative latency-associated gammaherpesvirus proteins and how vaccination may improve control of these viruses.
- Published
- 2004
108. Identification of a region of the virus genome involved in murine gammaherpesvirus 68-induced splenic pathology
- Author
-
Babunilayam Gangadharan, Stacey Efstathiou, Anthony Nash, James P. Stewart, Bahram Ebrahimi, Bernadette M. Dutia, and Douglas Roy
- Subjects
Rhadinovirus ,medicine.medical_specialty ,Pathology ,Anatomical pathology ,Spleen ,Genome, Viral ,Biology ,biology.organism_classification ,Virology ,Virus ,Mice ,Latent Virus ,medicine.anatomical_structure ,Interferon ,Mediastinal lymph node ,medicine ,Animals ,Gene ,Receptors, Interferon ,medicine.drug - Abstract
Infection with the murine gammaherpesvirus MHV-68 has profound effects on splenic and mediastinal lymph node pathology in mice which lack the interferon-γ receptor (IFN-γ R−/−). In these mice MHV-68 infection causes fibrosis and loss of lymphocytes in the spleen and the mediastinal lymph node as well as interstitial pulmonary fibrosis and fibrotic changes in the liver. The changes are associated with transient elevated latent virus loads in the spleen. Four independent virus mutants with insertions and/or deletions in the left end of the genome fail to induce the pathological changes and establish latency at normal levels in the spleen. The data indicate that the pathology does not correlate with any of the known genes encoded within this region of the genome, genes M1–M4 and the eight vtRNAs. Northern analysis of mRNAs transcribed by wild-type and mutant viruses shows that at least two uncharacterized transcripts are encoded within this region. These transcripts are absent in the mutant viruses and are candidates for the virus genes responsible for the aberrant pathology in IFN-γ R−/− mice.
- Published
- 2004
109. Correlation of TACC3, FGFR3, MMSET and p21 expression with the t(4;14)(p16·3;q32) in multiple myeloma
- Author
-
Terence R.J. Lappin, James P. Stewart, John D. Shaughnessy, Bart Barlogie, Alexander Thompson, and Madhumita Santra
- Subjects
Regulation of gene expression ,Candidate gene ,Real-time polymerase chain reaction ,Fusion transcript ,Breakpoint ,Gene expression ,Cancer research ,medicine ,Chromosomal translocation ,Hematology ,Biology ,medicine.disease ,Multiple myeloma - Abstract
The t(4;14)(p16;q32) translocation seen in c. 18% of newly diagnosed multiple myeloma (MM) cases, results in FGFR3 activation and creation of an IGH/MMSET fusion transcript. We have recently shown that FGFR3 is activated in only 75% of t(4;14)(+) cases, suggesting that alternative genes near the breakpoint may be involved in the transforming event. The gene, TACC3, located just 50 kb telomeric of FGFR3, with transforming capacity, therefore represented a candidate gene. Using a real-time quantitative polymerase chain reaction-based approach on a cohort of 54 patients, we found a statistically significant, twofold increase in TACC3 expression in t(4;14)(+) cases. TACC3, MMSET and p21 values were positively correlated in all cases and, of particular interest, six patient samples [three t(4;14)(-), three t(4;14)(+)] samples showed a joint up-regulation of TACC3, MMSET and p21. Although a poor prognosis is linked with elevated MMSET expression, an extended follow-up period will be required to evaluate the significance of elevated TACC3 and p21 expression in this subgroup of MM.
- Published
- 2004
110. Differential activation of murine herpesvirus 68- and Kaposi's sarcoma-associated herpesvirus-encoded ORF74 G protein-coupled receptors by human and murine chemokines
- Author
-
James P. Stewart, Carlos P. Fitzsimons, Marie van Dijk, Rob Leurs, Dennis Verzijl, Martine J. Smit, Henk Timmerman, Laboratory Medicine, ACS - Atherosclerosis & ischemic syndromes, Amsterdam Reproduction & Development (AR&D), Medicinal chemistry, and Chemistry and Pharmaceutical Sciences
- Subjects
Rhadinovirus ,Chemokine ,viruses ,Immunology ,medicine.disease_cause ,Microbiology ,Receptors, G-Protein-Coupled ,Mice ,Viral Proteins ,Chemokine receptor ,SDG 3 - Good Health and Well-being ,Virology ,Cyclic AMP ,medicine ,Animals ,Humans ,Gammaherpesvirinae ,CXCL10 ,Kaposi's sarcoma-associated herpesvirus ,G protein-coupled receptor ,biology ,NF-kappa B ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Molecular biology ,Virus-Cell Interactions ,Chemokine CXCL10 ,Type C Phospholipases ,Insect Science ,COS Cells ,Herpesvirus 8, Human ,biology.protein ,Receptors, Chemokine ,Chemokines ,Signal transduction ,Chemokines, CXC ,Protein Binding ,Signal Transduction - Abstract
Infection of mice with murine gammaherpesvirus 68 (MHV-68) is a well-characterized small animal model for the study of gammaherpesvirus infection. MHV-68 belongs to the same herpesvirus family as herpesvirus saimiri (HVS) of New World squirrel monkeys and human herpesvirus 8 (HHV-8) (also referred to as Kaposi's sarcoma-associated herpesvirus [KSHV]). The open reading frame ORF74 of HVS, KSHV, and MHV-68 encodes a protein with homology to G protein-coupled receptors and chemokine receptors in particular. ORF74 of KSHV (human ORF74 [hORF74]) is highly constitutively active and has been implicated in the pathogenesis of Kaposi's sarcoma. MHV-68-encoded ORF74 (mORF74) is oncogenic and has been implicated in viral replication and reactivation from latency. Here, we show that mORF74 is a functional chemokine receptor. Chemokines with an N-terminal glutamic acid-leucine-arginine (ELR) motif (e.g., KC and macrophage inflammatory protein 2) act as agonists on mORF74, activating phospholipase C, NF-κB, p44/p42 mitogen-activated protein kinase, and Akt signaling pathways and inhibiting formation of cyclic AMP. Using 125 I-labeled CXCL1/growth-related oncogene α as a tracer, we show that murine CXCL10/gamma interferon-inducible protein 10 binds mORF74, and functional assays show that it behaves as an antagonist for this virally encoded G protein-coupled receptor. Profound differences in the upstream activation of signal transduction pathways between mORF74 and hORF74 were found. Moreover, in contrast to hORF74, no constitutive activity of mORF74 could be detected.
- Published
- 2004
111. Ovine herpesvirus 2 lytic cycle replication and capsid production
- Author
-
Hugh W. Reid, Robert G. Dalziel, Jane Rosbottom, and James P. Stewart
- Subjects
T-Lymphocytes ,T cell ,Biology ,Virus Replication ,Virus ,Cell Line ,Capsid ,Gammaherpesvirinae ,Virology ,Gene expression ,medicine ,Animals ,Sheep ,Herpesviridae Infections ,biology.organism_classification ,Molecular biology ,medicine.anatomical_structure ,Viral replication ,Lytic cycle ,Cell culture ,DNA, Viral ,Cattle ,Rabbits ,DNA, Circular - Abstract
Ovine herpesvirus 2 (OvHV-2) causes malignant catarrhal fever in cattle, pigs and deer. We have observed intact circular and linear OvHV-2 genomes in infected T cell lines derived from cows and rabbits. Bovine T cell lines were predominantly latently infected but rabbit T cell lines supported OvHV-2 productive cycle gene expression and virus capsids were demonstrated for the first time.
- Published
- 2002
112. Murine gammaherpes virus as a cofactor in the development of pulmonary fibrosis in bleomycin resistant mice
- Author
-
Dcj Howell, James P. Stewart, S.S. Lok, Philip S. Hasleton, Jim J. Egan, and Y. Haider
- Subjects
Pulmonary and Respiratory Medicine ,Pathology ,medicine.medical_specialty ,Pulmonary Fibrosis ,Drug Resistance ,Bleomycin ,medicine.disease_cause ,Herpesviridae ,Virus ,Mice ,chemistry.chemical_compound ,Idiopathic pulmonary fibrosis ,Gammaherpesvirinae ,Fibrosis ,Pulmonary fibrosis ,Animals ,Medicine ,Lung ,Chromatography, High Pressure Liquid ,Mice, Inbred BALB C ,business.industry ,Respiratory disease ,Herpesviridae Infections ,respiratory system ,medicine.disease ,respiratory tract diseases ,Hydroxyproline ,medicine.anatomical_structure ,chemistry ,Collagen ,business - Abstract
Studies of human tissue have suggested an association between productive Epstein Barr virus and idiopathic pulmonary fibrosis (IPF). However, a pathogenic role for the virus has not been established. This study was undertaken to develop an animal model, which would explore the association between viral infection and pulmonary fibrosis. BALB/c mice (n=30), resistant to bleomycin, were primed with murine gammaherpesvirus 68 and then given intraperitoneal bleomycin. The mice were sacrificed at 28 days after bleomycin and their lungs assessed histologically and biochemically. Lung pathology was scored 0-3 for fibrotic and inflammatory change. BALB/c mice given virus and bleomycin showed more lung fibrosis (median score 2.2) compared to those given bleomycin alone (median 0), virus alone (median 0.2) or phosphate-buffered saline (PBS) control (median 0). Similarly mice given both virus and bleomycin showed more lung inflammation (median score 1.9) compared to those given bleomycin (median 0.5), virus (median 0.8), or PBS control (median 0.2). There was a significant difference in collagen content between the bleomycin and virus group (mean 1.86 mg) compared to the belomycin alone group (mean 1.52 mg). These results suggest that virus alone does not result in pulmonary fibrosis but that replicating virus in the presence of an exogenous injury may promote the development of pulmonary fibrosis.
- Published
- 2002
113. A Rearranged Form of Epstein–Barr Virus DNA Is Associated with Idiopathic Pulmonary Fibrosis
- Author
-
Jim J. Egan, She S. Lok, James P. Stewart, Brian G. Kelly, and Philip S. Hasleton
- Subjects
Male ,Pulmonary and Respiratory Medicine ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Pathology ,medicine.medical_specialty ,Pulmonary Fibrosis ,Buffy coat ,Virus Replication ,Critical Care and Intensive Care Medicine ,medicine.disease_cause ,Polymerase Chain Reaction ,Herpesviridae ,Viral Proteins ,Idiopathic pulmonary fibrosis ,Fibrosis ,Pulmonary fibrosis ,medicine ,Humans ,Epstein–Barr virus infection ,Gene Rearrangement ,Recombination, Genetic ,Lung ,business.industry ,Respiratory disease ,Chromosome Mapping ,Middle Aged ,respiratory system ,medicine.disease ,respiratory tract diseases ,DNA-Binding Proteins ,Blotting, Southern ,medicine.anatomical_structure ,Case-Control Studies ,DNA, Viral ,Immunology ,Trans-Activators ,Female ,business ,Immunosuppressive Agents ,Lung Transplantation - Abstract
An association between idiopathic pulmonary fibrosis (IPF) and productive Epstein-Barr virus (EBV) infection has been found previously. Productive EBV replication can be associated with a rearrangement in EBV genomes termed WZhet. We hypothesized that WZhet genomes might be present in patients with IPF. Thirty-nine patients with IPF, 26 lung transplant recipients, and 24 normal subjects were studied. When EBV DNA-positive lung tissue biopsies from IPF patients were analyzed, 11 of 18 (61%) were positive for WZhet. Buffy coat DNA analysis showed that 75-85% were EBV DNA-positive in both IPF and control groups. Buffy coat analysis for WZhet was positive in 16 of 27 (59%) IPF patients, compared with none of 32 lung transplant recipients and 1 of 24 (4%) normal blood donors (p < or = 0.001). There was thus a good correlation between the presence of WZhet in lung tissue and peripheral blood. However, there was no significant association between the presence of WZhet and immunosuppressive therapy. These data further confirm the association between active EBV infection and IPF and provide a potential marker in the peripheral blood for the tracking of EBV in this disease.
- Published
- 2002
114. Identification of Equid herpesvirus 2 in tissue-engineered equine tendon
- Author
-
Jane A. Pullman, Mohammed Al-Saadi, James P. Stewart, Sam Haldenby, Peter D. Clegg, Roisin Wardle, M D Pope, Alan D Radford, Lorenzo Ressel, Philip Dyer, and Mandy J. Peffers
- Subjects
0301 basic medicine ,Veterinary medicine ,040301 veterinary sciences ,viruses ,Medicine (miscellaneous) ,General Biochemistry, Genetics and Molecular Biology ,Virus ,DNA sequencing ,0403 veterinary science ,03 medical and health sciences ,0302 clinical medicine ,Retrovirus ,In vivo ,Virology ,Medicine ,Equine herpesvirus 2 ,tissue-engineered tendon ,superficial digital flexor tendon ,equine ,030304 developmental biology ,030203 arthritis & rheumatology ,0303 health sciences ,biology ,Contig ,business.industry ,Articles ,04 agricultural and veterinary sciences ,biology.organism_classification ,medicine.disease ,3. Good health ,Tendon ,030104 developmental biology ,medicine.anatomical_structure ,next-generation sequencing ,Tendinopathy ,business ,Research Article - Abstract
Background:Incidental findings of virus-like particles were identified following electron microscopy of tissue-engineered tendon constructs (TETC) derived from equine tenocytes. We set out to determine the nature of these particles, as there are few studies which identify virus in tendonsper se, and their presence could have implications for tissue-engineering using allogenic grafts.Methods:Virus particles were identified in electron microscopy of TETCs. Virion morphology was used to initially hypothesise the virus identity. Next generation sequencing was implemented to identify the virus. A pan herpesvirus PCR was used to validate the RNASeq findings using an independent platform. Histological analysis and biochemical analysis was undertaken on the TETCs.Results:Morphological features suggested the virus to be either a retrovirus or herpesvirus. Subsequent next generation sequencing mapped reads to Equid herpesvirus 2 (EHV2). Histological examination and biochemical testing for collagen content revealed no significant differences between virally affected TETCs and non-affected TETCs. An independent set of equine superficial digital flexor tendon tissue (n=10) examined using designed primers for specific EHV2 contigs identified at sequencing were negative. These data suggest that EHV is resident in some equine tendon.Conclusions:EHV2 was demonstrated in equine tenocytes for the first time; likely fromin vivoinfection. The presence of EHV2 could have implications to both tissue-engineering and tendinopathy.
- Published
- 2017
115. Analysis of the genetic diversity of ovine herpesvirus 2 in samples from livestock with malignant catarrhal fever
- Author
-
George C. Russell, Bruce Lamond, David F. Twomey, A. E. Courtenay, Kim Willoughby, David M. Haig, James P. Stewart, David Norris, Dawn M. Grant, and Sandra Scholes
- Subjects
Rhadinovirus ,Genes, Viral ,Sequence analysis ,Molecular Sequence Data ,Sheep Diseases ,Biology ,Microbiology ,DNA sequencing ,Virus ,Polymorphism (computer science) ,Animals ,Amino Acid Sequence ,Allele ,Gene ,Alleles ,Phylogeny ,Genetics ,Genetic diversity ,Sheep ,General Veterinary ,Genetic Variation ,General Medicine ,Virology ,Malignant Catarrh ,RNA splicing ,Cattle - Abstract
In order to define better virus isolates from animals with malignant catarrhal fever (MCF), segments of three genes of ovine herpesvirus-2 were amplified from diagnostic samples representing MCF cases with a range of clinical presentations in cattle, including head and eye, alimentary and neurological. The variation within each gene segment was estimated by DNA sequencing, which confirmed that the newly-annotated Ov9.5 gene was significantly more polymorphic than either of the other loci tested (segments of ORF50 and ORF75), with alleles that differed at over 60% of nucleotide positions. Despite this, the nine Ov9.5 alleles characterised had identical predicted splicing patterns and could be translated into Ov9.5 polypeptides with at least 49% amino acid identity. This multi-locus approach has potential for use in epidemiological studies and in charactering chains of infection. However there was no association between specific variants of OvHV-2 and the clinical/pathological presentation of MCF in the cattle analysed.
- Published
- 2014
116. Latent Antigen Vaccination in a Model Gammaherpesvirus Infection
- Author
-
David L. Woodland, Marcia A. Blackman, Kimberley A. Ward, Edward J. Usherwood, and James P. Stewart
- Subjects
Immunology ,Epitopes, T-Lymphocyte ,CD8-Positive T-Lymphocytes ,Microbiology ,Epitope ,Viral Matrix Proteins ,Mice ,Gammaherpesvirinae ,H-2 Antigens ,Antigen ,Virology ,Vaccines and Antiviral Agents ,Virus latency ,Vaccines, DNA ,medicine ,Animals ,Humans ,Latency (engineering) ,Antigens, Viral ,Lung ,Mice, Inbred BALB C ,biology ,Viral Vaccine ,Vaccination ,Viral Vaccines ,Herpesviridae Infections ,biology.organism_classification ,medicine.disease ,Virus Latency ,Disease Models, Animal ,Insect Science - Abstract
Vaccines that can reduce the load of latent gammaherpesvirus infections are eagerly sought. One attractive strategy is vaccination against latency-associated proteins, which may increase the efficiency with which T cells recognize and eliminate latently infected cells. However, due to the lack of tractable animal model systems, the effect of latent-antigen vaccination on gammaherpesvirus latency is not known. Here we use the murine gammaherpesvirus model to investigate the impact of vaccination with the latency-associated M2 antigen. As expected, vaccination had no effect on the acute lung infection. However, there was a significant reduction in the load of latently infected cells in the initial stages of the latent infection, when M2 is expressed. These data show for the first time that latent-antigen vaccination can reduce the level of latency in vivo and suggest that vaccination strategies involving other latent antigens may ultimately be successfully used to reduce the long-term latent infection.
- Published
- 2001
117. Characterization of the murine gammaherpesvirus 68 ORF74 product: a novel oncogenic G protein-coupled receptor
- Author
-
Madeleine N. Wakeling, James P. Stewart, Anthony Nash, and Douglas Roy
- Subjects
Transcription, Genetic ,viruses ,Molecular Sequence Data ,Gene Expression ,Receptors, Cell Surface ,Biology ,Cell Line ,Mice ,Open Reading Frames ,Viral Proteins ,Gammaherpesvirinae ,GTP-binding protein regulators ,GTP-Binding Proteins ,Transcription (biology) ,Cricetinae ,Virology ,Animals ,Amino Acid Sequence ,Peptide sequence ,Gene ,G protein-coupled receptor ,Oncogene Proteins ,Mice, Inbred BALB C ,Gene Expression Profiling ,virus diseases ,3T3 Cells ,biochemical phenomena, metabolism, and nutrition ,Cell Transformation, Viral ,Open reading frame ,Transmembrane domain ,Female ,Sequence motif ,hormones, hormone substitutes, and hormone antagonists - Abstract
Murine gammaherpesvirus (MHV-68) is well established as a small animal model for the study of gammaherpesviruses. The MHV-68 genome contains an open reading frame (ORF74) that has significant sequence homology with mammalian G-protein coupled receptors (GPCRs) and the GPCR from the related Kaposi’s sarcoma-associated herpesvirus (KSHV). Here we show that the MHV-68 ORF74 is predicted to encode a GPCR since it has seven potential transmembrane helices and that it has other sequence motifs in common with GPCRs. Of interest is the observation that the sequence around a conserved arginine at the start of the second intracellular loop suggests that the ORF74 product may signal constitutively (agonist independent). Given that the ORF74 product is predicted to encode a GPCR we named it MHV-GPCR. In studies on the transcription of the MHV-GPCR, we determined that it was encoded on multiple early transcripts of 3·4, 4·4, 6·6 and 8·7 kb in size. At least one of these transcripts was bicistronic, containing the ORF encoding the Bcl-2 homologue also.In vivo, we found that MHV GPCR was expressed during acute infection but also during persistence, particularly in the lungs of infected mice. Immunofluorescence studies indicated that the MHV GPCR protein was expressed on the surface of cells in patches. Finally, like the KSHV GPCR, expression of the MHV GPCR resulted in transformation of NIH 3T3 cells. We surmise, therefore, that the MHV GPCR may act in concert with genes with which it is expressed such as vBcl-2 to enhance the growth and survival of MHV-68-infected cells.
- Published
- 2001
118. Natural history of murine γ-herpesvirus infection
- Author
-
James P. Stewart, Anthony Nash, Bernadette M. Dutia, and Andrew J. Davison
- Subjects
Lymphoid Tissue ,Genome, Viral ,Biology ,Major histocompatibility complex ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Immunocompromised Host ,Mice ,Gammaherpesvirinae ,Immune system ,Latent Virus ,Interferon ,Virus latency ,medicine ,Animals ,Humans ,Lung ,Virulence ,Vaccination ,Herpesviridae Infections ,medicine.disease ,Virology ,Lymphoproliferative Disorders ,Virus Latency ,Tumor Virus Infections ,Immunity, Active ,Viral replication ,Immunology ,biology.protein ,General Agricultural and Biological Sciences ,CD8 ,medicine.drug - Abstract
Murine γ–herpesvirus 68 (MHV–68) is a natural pathogen of small rodents and insectivores (mice, voles and shrews). The primary infection is characterized by virus replication in lung epithelial cells and the establishment of a latent infection in B lymphocytes. The virus is also observed to persist in lung epithelial cells, dendritic cells and macrophages. Splenomegaly is observed two weeks after infection, in which there is a CD4 + T–cell–mediated expansion of B and T cells in the spleen. At three weeks post–infection an infectious mononucleosis–like syndrome is observed involving a major expansion of Vβ4 + CD8 + T cells. Later in the course of persistent infection, ca . 10% of mice develop lymphoproliferative disease characterized as lymphomas of B–cell origin. The genome from MHV–68 strain g2.4 has been sequenced and contains ca . 73 genes, the majority of which are collinear and homologous to other γ–herpesviruses. The genome includes cellular homologues for a complement–regulatory protein, Bcl–2, cyclin D and interleukin–8 receptor and a set of novel genes M1 to M4. The function of these genes in the context of latent infections, evasion of immune responses and virus–mediated pathologies is discussed. Both innate and adaptive immune responses play an active role in limiting virus infection. The absence of type I interferon (IFN) results in a lethal MHV–68 infection, emphasizing the central role of these cytokines at the initial stages of infection. In contrast, type II IFN is not essential for the recovery from infection in the lung, but a failure of type II IFN receptor signalling results in the atrophy of lymphoid tissue associated with virus persistence. Splenic atrophy appears to be the result of immunopathology, since in the absence of CD8 + T cells no pathology occurs. CD8 + T cells play a major role in recovery from the primary infection, and also in regulating latently infected cells expressing the M2 gene product. CD4 + T cells have a key role in surveillance against virus recurrences in the lung, in part mediated through ‘help’ in the genesis of neutralizing antibodies. In the absence of CD4 + T cells, virus–specific CD8 + T cells are able to control the primary infection in the respiratory tract, yet surprisingly the memory CD8 + T cells generated are unable to inhibit virus recurrences in the lung. This could be explained in part by the observations that this virus can downregulate major histocompatibility complex class I expression and also restrict inflammatory cell responses by producing a chemokine–binding protein (M3 gene product). MHV–68 provides an excellent model to explore methods for controlling γ–herpesvirus infection through vaccination and chemotherapy. Vaccination with gp150 (a homologue of gp350 of Epstein–Barr virus) results in a reduction in splenomegaly and virus latency but does not block replication in the lung, nor the establishment of a latent infection. Even when lung virus infection is greatly reduced following the action of CD8 + T cells, induced via a prime–boost vaccination strategy, a latent infection is established. Potent antiviral compounds such as the nucleoside analogue 2′deoxy–5–ethyl–beta–4′–thiouridine, which disrupts virus replication in vivo , cannot inhibit the establishment of a latent infection. Clearly, devising strategies to interrupt the establishment of latent virus infections may well prove impossible with existing methods.
- Published
- 2001
119. Comparison of transfemoral transcatheter aortic valve replacement performed in the catheterization laboratory (minimalist approach) versus hybrid operating room (standard approach): outcomes and cost analysis
- Author
-
Vasilis, Babaliaros, Chandan, Devireddy, Stamatios, Lerakis, Robert, Leonardi, Sebastian A, Iturra, Kreton, Mavromatis, Bradley G, Leshnower, Robert A, Guyton, Mihir, Kanitkar, Patricia, Keegan, Amy, Simone, James P, Stewart, Nima, Ghasemzadeh, Peter, Block, and Vinod H, Thourani
- Subjects
Aged, 80 and over ,Heart Valve Prosthesis Implantation ,Male ,Cardiac Catheterization ,Operating Rooms ,Georgia ,Time Factors ,Cost-Benefit Analysis ,Process Assessment, Health Care ,Aortic Valve Stenosis ,Anesthesia, General ,Length of Stay ,Prosthesis Design ,Femoral Artery ,Intensive Care Units ,Treatment Outcome ,Cost Savings ,Risk Factors ,Heart Valve Prosthesis ,Humans ,Female ,Hospital Costs ,Echocardiography, Transesophageal ,Aged ,Retrospective Studies - Abstract
The aim of this study was to compare transfemoral transcatheter aortic valve replacement (TF TAVR) performed in a catheterization laboratory (minimalist approach [MA]) with TF TAVR performed in a hybrid operating room (standard approach [SA]).A MA-TF TAVR can be performed without general anesthesia, transesophageal echocardiography, or a surgical hybrid room. The outcomes and cost of MA-TF TAVR compared with those of the SA have not been described.Patients who underwent elective, percutaneous TF TAVR using the Edwards Sapien valve (Edwards Lifesciences, Irvine, California) were studied. Baseline characteristics, outcomes, and hospital costs of MA-TF TAVR and SA-TF TAVR were compared.A total of 142 patients were studied (MA-TF TAVR, n = 70 and SA-TF TAVR, n = 72). There were no differences in baseline comorbidities (Society of Thoracic Surgeons score, 10.6 ± 4.3 vs. 11.4 ± 5.8; p = 0.35). All procedures in the MA-TF TAVR group were successful; 1 patient was intubated. Three patients in the SA-TF TAVR group had procedure-related death. Procedure room time (150 ± 48 min vs. 218 ± 56 min, p0.001), total intensive care unit time (22 h vs. 28 h, p0.001), length of stay from procedure to discharge (3 days vs. 5 days, p0.001), and cost ($45,485 ± 14,397 vs. $55,377 ± 22,587, p0.001) were significantly less in the MA-TF TAVR group. Mortality at 30 days was not significantly different in the MA-TF TAVR group (0 vs. 6%, p = 0.12) and 30-day stroke/transient ischemic attack was similar (4.3% vs. 1.4%, p = 0.35). Moderate or severe paravalvular leak and device success were similar in the MA-TF TAVR and SA-TF TAVR groups (3% vs. 5.8%, p = 0.4 and 90% vs. 88%, p = 0.79, respectively) at 30 days. At a median follow-up of 435 days, there was no significant difference in survival (MA-TF TAVR, 83% vs. SA-TF TAVR, 82%; p = 0.639).MA-TF TAVR can be performed with minimal morbidity and mortality and equivalent effectiveness compared with SA-TF TAVR. The shorter length of stay and lower resource use with MA-TF TAVR significantly lowers hospital costs.
- Published
- 2013
120. Rta of Murine Gammaherpesvirus 68 Reactivates the Complete Lytic Cycle from Latency
- Author
-
Anthony Nash, Edward J. Usherwood, Ting-Ting Wu, James P. Stewart, and Ren Sun
- Subjects
Gene Expression Regulation, Viral ,Transcription, Genetic ,viruses ,Immunology ,Replication ,Viral Plaque Assay ,Transfection ,Virus Replication ,Microbiology ,Immediate early protein ,Virus ,Immediate-Early Proteins ,Mice ,Viral Proteins ,Gammaherpesvirinae ,Plasmid ,Virology ,Virus latency ,medicine ,Animals ,Gene ,Cells, Cultured ,Cell Line, Transformed ,B-Lymphocytes ,biology ,Virion ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,medicine.disease ,Virus Latency ,Lytic cycle ,Viral replication ,Insect Science ,Trans-Activators ,Virus Activation - Abstract
Herpesviruses are characterized as having two distinct life cycle phases: lytic replication and latency. The mechanisms of latency establishment and maintenance, as well as the switch from latency to lytic replication, are poorly understood. Human gammaherpesviruses, including Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), are associated with lymphoproliferative diseases and several human tumors. Unfortunately, the lack of cell lines to support efficient de novo productive infection and restricted host ranges of EBV and HHV-8 make it difficult to explore certain important biological questions. Murine gammaherpesvirus 68 (MHV-68, or γHV68) can establish de novo lytic infection in a variety of cell lines and is also able to infect laboratory mice, offering an ideal model with which to study various aspects of gammaherpesvirus infection. Here we describe in vitro studies of the mechanisms of the switch from latency to lytic replication of MHV-68. An MHV-68 gene, rta (replication and transcription activator), encoded primarily by open reading frame 50 (ORF50), is homologous to the rta genes of other gammaherpesviruses, including HHV-8 and EBV. HHV-8 and EBV Rta have been shown to play central roles in viral reactivation from latency. We first studied the kinetics of MHV-68 rta gene transcription during de novo lytic infection. MHV-68 rta was predominantly expressed as a 2-kb immediate-early transcript. Sequence analysis of MHV-68 rta cDNA revealed that an 866-nucleotide intron 5′ of ORF50 was removed to create the Rta ORF of 583 amino acids. To test the functions of MHV-68 Rta in reactivation, a plasmid expressing Rta was transfected into a latently infected cell line, S11E, which was established from a B-cell lymphoma in an MHV-68-infected mouse. Rta induced expression of viral early and late genes, lytic replication of viral DNA, and production of infectious viral particles. We conclude that Rta alone is able to disrupt latency, activate viral lytic replication, and drive the lytic cycle to completion. This study indicates that MHV-68 provides a valuable model for investigating regulation of the balance between latency and lytic replication in vitro and in vivo.
- Published
- 2000
121. Control of Gammaherpesvirus Latency by Latent Antigen-Specific Cd8+ T Cells
- Author
-
Marcia A. Blackman, Kim Ward, Douglas Roy, Bernadette M. Dutia, Sherri L. Surman, David L. Woodland, Edward J. Usherwood, and James P. Stewart
- Subjects
Adoptive cell transfer ,Genes, Viral ,T cell ,Immunology ,Epitopes, T-Lymphocyte ,Biology ,CD8-Positive T-Lymphocytes ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Gammaherpesvirinae ,Antigen ,Virus latency ,medicine ,Tumor Cells, Cultured ,Immunology and Allergy ,Cytotoxic T cell ,Animals ,Humans ,Antigens, Viral ,B cell ,030304 developmental biology ,0303 health sciences ,B-Lymphocytes ,Mice, Inbred BALB C ,gammaherpesvirus ,viral antigens ,Gene Expression Profiling ,H-2 Antigens ,CD8+ T lymphocytes ,medicine.disease ,Virology ,3. Good health ,viral load ,Virus Latency ,Kinetics ,medicine.anatomical_structure ,Original Article ,Immunologic Memory ,CD8 ,030215 immunology - Abstract
The contribution of the latent antigen-specific CD8+ T cell response to the control of gammaherpesvirus latency is currently obscure. Some latent antigens induce potent T cell responses, but little is known about their induction or the role they play during the establishment of latency. Here we used the murine gammaherpesvirus system to examine the expression of the latency-associated M2 gene during latency and the induction of the CD8+ T cell response to this protein. M2, in contrast to the M3 latency-associated antigen, was expressed at day 14 after infection but was undetectable during long-term latency. The induction of the M291–99/Kd CD8+ T cell response was B cell dependent, transient, and apparently induced by the rapid increase in latently infected cells around day 14 after intranasal infection. These kinetics were consistent with a role in controlling the initial “burst” of latently infected cells. In support of this hypothesis, adoptive transfer of an M2-specific CD8+ T cell line reduced the initial load of latently infected cells, although not the long-term load. These data represent the first description of a latent antigen-specific immune response in this model, and suggest that vaccination with latent antigens such as M2 may be capable of modulating latent gammaherpesvirus infection.
- Published
- 2000
122. The Detection of Epstein-Barr Virus DNA in Lung Tissue from Patients with Idiopathic Pulmonary Fibrosis
- Author
-
James P. Stewart, She S. Lok, Ashley Woodcock, Philip S. Hasleton, Alan J. Ross, Brian G. Kelly, and Jim J. Egan
- Subjects
Adult ,Male ,Pulmonary and Respiratory Medicine ,Herpesvirus 4, Human ,Pathology ,medicine.medical_specialty ,Pulmonary Fibrosis ,Critical Care and Intensive Care Medicine ,medicine.disease_cause ,Polymerase Chain Reaction ,Herpesviridae ,Serology ,Viral Matrix Proteins ,Idiopathic pulmonary fibrosis ,Capsid ,Fibrosis ,hemic and lymphatic diseases ,Humans ,Medicine ,Gammaherpesvirinae ,Antigens, Viral ,Lung ,Aged ,biology ,business.industry ,Middle Aged ,respiratory system ,medicine.disease ,biology.organism_classification ,Immunohistochemistry ,Epstein–Barr virus ,respiratory tract diseases ,DNA, Viral ,Immunology ,Capsid Proteins ,Female ,business ,Nested polymerase chain reaction - Abstract
Idiopathic pulmonary fibrosis (IPF) is a clinical syndrome in which the precipitating factors are unclear. An association between Epstein-Barr Virus (EBV) and IPF had previously been suggested using serology and immunohistochemistry. This study sought confirmation of the presence of EBV DNA in the lung tissue of patients with IPF. Lung tissue obtained surgically from 27 patients with IPF and 28 control subjects was investigated for the presence of EBV by immunohistochemistry and polymerase chain reaction (PCR) analysis. Immunohistochemistry used antibodies specific for EBV lytic cycle antigens (gp340/220 and VCA). Nested PCR analysis used oligonucleotide primers specific for EBV and was sensitive to one copy of EBV DNA. Twelve of the 27 patients with IPF (44%) and three of the 28 control subjects (10%) were EBV positive by immunohistochemistry (p = 0.005). Thirteen of the patients with IPF (48%) and four of the control subjects (14%) were EBV positive by PCR (p = 0.007). Eleven of the patients with IPF (41%) and none of the control subjects were EBV positive by both immunohistochemistry and PCR (p = < 0.001). These data further suggest an association between EBV and IPF. In addition it defines a novel method for detecting EBV in lung tissue. EBV may be involved in the pathogenesis of the disease; however, further studies are required to establish a causal relationship.
- Published
- 1999
123. Lung Epithelial Cells Are a Major Site of Murine Gammaherpesvirus Persistence
- Author
-
Edward J. Usherwood, Heather Dyson, Tony Nash, James P. Stewart, and Alan J. Ross
- Subjects
Adoptive cell transfer ,viruses ,Immunology ,Mice, Transgenic ,Spleen ,In situ hybridization ,Biology ,medicine.disease_cause ,Polymerase Chain Reaction ,Virus ,Mice ,Gammaherpesvirinae ,Virus latency ,medicine ,Animals ,Epstein-Barr virus ,Immunology and Allergy ,Lung ,In Situ Hybridization ,latency ,lungs ,gammaherpesvirus ,Immunoglobulin mu-Chains ,Murid herpesvirus 4 ,Virus Activation ,epithelia ,Epithelial Cells ,Herpesviridae Infections ,Articles ,medicine.disease ,biology.organism_classification ,Epstein–Barr virus ,Virology ,Virus Latency ,medicine.anatomical_structure ,DNA, Viral ,Plasmids - Abstract
It is currently believed that latently infected, resting B lymphocytes are central to gammaherpesvirus persistence, whereas mucosal epithelial cells are considered nonessential. We have readdressed the question of nonlymphoid persistence using murine gammaherpesvirus 68 (MHV-68). To dissect lymphoid from nonlymphoid persistence, we used μMT transgenic mice that are defective in B cells. MHV-68 DNA persisted in the lungs of intact and B cell–deficient mice. Both episomal and linear forms of the virus genome were present in lungs, implying the presence of both latency and productive replication. In situ hybridization for virus tRNA transcripts revealed latent MHV-68 in pulmonary epithelial cells. Infectious virus was recovered from the lungs of μMT mice after T cell depletion, showing that the persisting virus DNA was reactivatable. Finally, using adoptive transfer of B cells into B cell–deficient mice, it was shown that virus persisting in lungs seeded splenic B cells, and virus resident in the spleen seeded the lungs. These results show that mucosal epithelia can act as a nonlymphoid reservoir for gammaherpesvirus persistence, and that there is a two-way movement of virus between lymphoid and nonlymphoid compartments during persistence.
- Published
- 1998
124. Viruses and idiopathic pulmonary fibrosis
- Author
-
Ashley Woodcock, James P. Stewart, and Jim J. Egan
- Subjects
Pulmonary and Respiratory Medicine ,Idiopathic pulmonary fibrosis ,Pathology ,medicine.medical_specialty ,Text mining ,business.industry ,Medicine ,business ,medicine.disease - Published
- 1997
125. Genetic content and preliminary transcriptional analysis of a representative region of murine gammaherpesvirus 68
- Author
-
James P. Stewart, Stacey Efstathiou, S de V Pepper, Mike Mackett, M Chee, J. R. Arrand, and Anthony Nash
- Subjects
Herpesvirus 4, Human ,Transcription, Genetic ,RNA Splicing ,viruses ,Molecular Sequence Data ,Rodentia ,Locus (genetics) ,Genome ,Virus ,Open Reading Frames ,Gammaherpesvirinae ,Transcription (biology) ,hemic and lymphatic diseases ,Virology ,Animals ,Northern blot ,Gene ,Genetics ,Base Sequence ,biology ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Open reading frame - Abstract
Murine gammaherpesvirus 68 (MHV-68) is a relatively recently discovered pathogen of wild rodents and provides a unique opportunity to explore in detail the interactions of a gammaherpesvirus with its natural host. It may also provide a much needed small animal model for human gammaherpesviruses. As a step in the detailed analysis of virus gene structure and expression we have sequenced over 20 kb of the MHV-68 genome and mapped gene transcripts by Northern blot hybridization. The region we chose to analyse contains several conserved gene blocks as well as some less well conserved genes and allowed us to estimate the relationship of this virus to other herpesvirus family members. Of particular interest is the fact that none of the characteristic Epstein-Barr virus (EBV) genes is present at this genomic locus although MHV-68 does have one gene encoding a membrane glycoprotein, 9p150, which shows similarities to the major membrane glycoprotein of EBV. Our results further confirm that MHV-68 is a gammaherpesvirus marginally more closely related to a cluster of gammaherpesviruses including herpesvirus salmiri than to EBV. Northern analysis shows that the temporal regulation of expression is broadly similar to that of other herpesviruses in this region of the genome. We also show that like other gammaherpesviruses, MHV-68 splices its homologue of the EBV transcriptional activator gene BMRF1.
- Published
- 1997
126. 'Unconscious migrants' discovered in van on ferry.
- Author
-
James Fielding ; Stewart Carr
- Abstract
HORRIFIED tourists told last night how unconscious migrants were pulled from the back of a van driven on to a ferry by suspected people traffickers. [ABSTRACT FROM PUBLISHER]
- Published
- 2024
127. Herpes Virus Infection Is Associated with Vascular Remodeling and Pulmonary Hypertension in Idiopathic Pulmonary Fibrosis
- Author
-
Emanuela Rossi, Federico Rea, James P. Stewart, Francesca Lunardi, Anja Kipar, Egle Perissinotto, Fiorella Calabrese, Giuseppe Marulli, Elisabetta Balestro, Nazarena Nannini, Departments of Faculty of Veterinary Medicine, Veterinary Biosciences, Veterinary Pathology and Parasitology, University of Zurich, and Calabrese, F
- Subjects
Male ,Viral Diseases ,Pulmonology ,viruses ,medicine.medical_treatment ,Veterinary Microbiology ,GAMMAHERPESVIRUS ,lcsh:Medicine ,Cardiovascular ,413 Veterinary science ,Idiopathic pulmonary fibrosis ,0302 clinical medicine ,Transforming Growth Factor beta ,Fibrosis ,Molecular Cell Biology ,Pulmonary fibrosis ,lcsh:Science ,EPSTEIN-BARR-VIRUS ,Lung ,Herpesviridae ,0303 health sciences ,Multidisciplinary ,MEMBRANE-PROTEIN 1 ,Herpesviridae Infections ,LUNG FIBROSIS ,Middle Aged ,respiratory system ,EPITHELIAL-MESENCHYMAL TRANSITION ,humanities ,3. Good health ,Infectious Diseases ,medicine.anatomical_structure ,Veterinary Diseases ,Hypertension ,Medicine ,Female ,Veterinary Pathology ,Research Article ,Lung Transplantation ,Histology ,Clinical Research Design ,Hypertension, Pulmonary ,education ,10184 Institute of Veterinary Pathology ,1100 General Agricultural and Biological Sciences ,Genome, Viral ,Interstitial Lung Diseases ,Pulmonary Artery ,Microbiology ,03 medical and health sciences ,1300 General Biochemistry, Genetics and Molecular Biology ,EBV ,medicine ,Animals ,Humans ,Lung transplantation ,Arterial Pressure ,Pulmonary pathology ,Biology ,030304 developmental biology ,1000 Multidisciplinary ,business.industry ,TRANSPLANTATION ,lcsh:R ,Herpes Simplex ,Epithelial Cells ,DNA ,medicine.disease ,Pulmonary hypertension ,Idiopathic Pulmonary Fibrosis ,respiratory tract diseases ,Transplantation ,Disease Models, Animal ,MICE ,030228 respiratory system ,Alveolar Epithelial Cells ,Immunology ,CELLS ,570 Life sciences ,biology ,Blood Vessels ,Veterinary Science ,lcsh:Q ,business - Abstract
Background Pulmonary hypertension (PH) represents an important complication of idiopathic pulmonary fibrosis (IPF) with a negative impact on patient survival. Herpes viruses are thought to play an etiological role in the development and/or progression of IPF. The influence of viruses on PH associated with IPF is unknown. We aimed to investigate the influence of viruses in IPF patients focusing on aspects related to PH. A laboratory mouse model of gamma-herpesvirus (MHV-68) induced pulmonary fibrosis was also assessed. Methods Lung tissue samples from 55 IPF patients and 41 controls were studied by molecular analysis to detect various viral genomes. Viral molecular data obtained were correlated with mean pulmonary arterial pressure (mPAP) and arterial remodelling. Different clinical and morphological variables were studied by univariate and multivariate analyses at time of transplant and in the early post-transplant period. The same lung tissue analyses were performed in MHV-68 infected mice. Results A higher frequency of virus positive cases was found in IPF patients than in controls (p = 0.0003) and only herpes virus genomes were detected. Viral cases showed higher mPAP (p = 0.01), poorer performance in the six minute walking test (6MWT; p = 0.002) and higher frequency of primary graft (PGD) dysfunction after lung transplant (p = 0.02). Increased arterial thickening, particularly of the intimal layer (p = 0.002 and p = 0.004) and higher TGF-β expression (p = 0.002) were demonstrated in viral cases. The remodelled vessels showed increased vessel cell proliferation (Ki-67 positive cells) in the proximity to metaplastic epithelial cells and macrophages. Viral infection was associated with higher mPAP (p = 0.03), poorer performance in the 6MWT (p = 0.008) and PGD (p = 0.02) after adjusting for other covariates/intermediate factors. In MHV-68 infected mice, morphological features were similar to those of patients. Conclusion Herpesviral infections may contribute to the development of PH in IPF patients.
- Published
- 2013
128. Role of Pituitary Adenylate-Cyclase Activating Polypeptide and Tac1 gene derived tachykinins in sensory, motor and vascular functions under normal and neuropathic conditions
- Author
-
Bálint Botz, Zsuzsanna Helyes, Andreas Zimmer, János Szolcsányi, Hitoshi Hashimoto, András Imreh, John P. Quinn, James P. Stewart, Krisztián Elekes, Katalin Sándor, and Dóra Reglődi
- Subjects
medicine.medical_specialty ,Sensory Receptor Cells ,Physiology ,Neuropeptide ,Neovascularization, Physiologic ,Substance P ,Calcitonin gene-related peptide ,Motor Activity ,Biochemistry ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Mice ,Endocrinology ,Internal medicine ,Tachykinins ,medicine ,Animals ,Mice, Knockout ,business.industry ,Receptors, Neurokinin-1 ,Mice, Inbred C57BL ,Nociception ,chemistry ,Neuropathic pain ,Pituitary Adenylate Cyclase-Activating Polypeptide ,Neurokinin A ,Sciatic nerve ,Tachykinin receptor ,business ,hormones, hormone substitutes, and hormone antagonists - Abstract
Pituitary Adenylate-Cyclase Activating Polypeptide (PACAP) and Tac1 gene-encoded tachykinins (substance P: SP, neurokinin A: NKA) are expressed in capsaicin-sensitive nerves, but their role in nociception, inflammation and vasoregulation is unclear. Therefore, we investigated the function of these neuropeptides and the NK1 tachykinin receptor (from Tacr1 gene) in the partial sciatic nerve ligation-induced traumatic mononeuropathy model using gene deficient (PACAP(-/-), Tac1(-/-), and Tacr1(-/-)) mice. Mechanonociceptive threshold of the paw was measured with dynamic plantar aesthesiometry, motor coordination with Rota-Rod and cutaneous microcirculation with laser Doppler imaging. Neurogenic vasodilation was evoked by mustard oil stimulating sensory nerves. In wildtype mice 30-40% mechanical hyperalgesia developed one week after nerve ligation, which was not altered in Tac1(-/-) and Tacr1(-/-) mice, but was absent in PACAP(-/-) animals. Motor coordination of the PACAP(-/-) and Tac1(-/-) groups was significantly worse both before and after nerve ligation compared to their wildtypes, but it did not change in Tacr1(-/-) mice. Basal postoperative microcirculation on the plantar skin of PACAP(-/-) mice did not differ from the wildtypes, but was significantly lower in Tac1(-/-) and Tacr1(-/-) ones. In contrast, mustard oil-induced neurogenic vasodilation was significantly smaller in PACAP(-/-) mice, but not in Tacr1(-/-) and Tac1(-/-) animals. Both PACAP and SP/NKA, but not NK1 receptors participate in normal motor coordination. Tachykinins maintain basal cutaneous microcirculation. PACAP is a crucial mediator of neuropathic mechanical hyperalgesia and neurogenic vasodilation. Therefore identifying its target and developing selective, potent antagonists, might open promising new perspectives for the treatment of neuropathic pain and vascular complications.
- Published
- 2013
129. PCR based bronchoscopic detection of common respiratory pathogens in chronic cough: a case control study
- Author
-
Paul W. Bishop, Ashley Woodcock, Jaclyn A. Smith, Samantha Decalmer, Peter W. West, James P. Stewart, Winifred Dove, and Angela Kelsall
- Subjects
Pulmonary and Respiratory Medicine ,Bronchoalveolar lavage ,medicine.medical_specialty ,Pathology ,Lymphocytosis ,Cough reflex ,Biopsy ,Gastroenterology ,Viral Respiratory Tract Infection ,Internal medicine ,Bronchoscopy ,medicine ,Lung ,medicine.diagnostic_test ,business.industry ,Research ,Case-control study ,Virus ,respiratory tract diseases ,Chronic cough ,medicine.anatomical_structure ,PCR ,Otorhinolaryngology ,Cough ,medicine.symptom ,business ,Infection - Abstract
Background Viral respiratory tract infection is the most frequent cause of acute cough and is reported at onset in about one third of patients with chronic cough. Persistent infection is therefore one possible explanation for the cough reflex hypersensitivity and pulmonary inflammation reported in chronic cough patients. Methods Bronchoscopic endobronchial biopsies and bronchoalveolar lavage cell counts were obtained from ten healthy volunteers and twenty treatment resistant chronic cough patients (10 selected for lavage lymphocytosis). A screen for known respiratory pathogens was performed on biopsy tissue. Chronic cough patients also underwent cough reflex sensitivity testing using citric acid. Results There was no significant difference in incidence of infection between healthy volunteers and chronic cough patients (p = 0.115) or non-lymphocytic and lymphocytic groups (p = 0.404). BAL cell percentages were not significantly different between healthy volunteers and chronic cough patients without lymphocytosis. Lymphocytic patients however had a significantly raised percentage of lymphocytes (p Conclusions This study indicates latent infection in the lung is unlikely to play an important role in chronic cough, but a role for undetected or undetectable pathogens in either the lung or a distal site could not be ruled out. Trials registration Current Controlled Trials ISRCTN62337037 & ISRCTN40147207
- Published
- 2013
- Full Text
- View/download PDF
130. Role of tachykinin 1 and 4 gene-derived neuropeptides and the neurokinin 1 receptor in adjuvant-induced chronic arthritis of the mouse
- Author
-
Éva Borbély, Alexandra Berger, István János Tóth, Péter Nagy, Zsófia Hajna, James P. Stewart, Zsuzsanna Helyes, János Szolcsányi, Katalin Sándor, Christopher J. Paige, Erika Pintér, László Kereskai, Andreas Zimmer, and John P. Quinn
- Subjects
Male ,Mouse ,Freund's Adjuvant ,Interleukin-1beta ,Arthritis ,lcsh:Medicine ,Substance P ,Autoimmunity ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Genetics of the Immune System ,Edema ,Receptor ,lcsh:Science ,Mice, Knockout ,0303 health sciences ,Neurogenic inflammation ,Multidisciplinary ,Animal Models ,Neurotransmitters ,Receptors, Neurokinin-1 ,Plethysmography ,Hyperalgesia ,Medicine ,medicine.symptom ,Research Article ,Signal Transduction ,Agonist ,medicine.medical_specialty ,medicine.drug_class ,Immunology ,Rheumatoid Arthritis ,Tarsus, Animal ,Autoimmune Diseases ,03 medical and health sciences ,Model Organisms ,Rheumatology ,Internal medicine ,Tachykinins ,Tachykinin receptor 1 ,medicine ,Genetics ,Animals ,Protein Precursors ,Biology ,030304 developmental biology ,Inflammation ,business.industry ,lcsh:R ,medicine.disease ,Arthritis, Experimental ,Mice, Inbred C57BL ,Endocrinology ,chemistry ,Gene Expression Regulation ,Freund's adjuvant ,Genetics of Disease ,Clinical Immunology ,Joints ,lcsh:Q ,Gene Function ,business ,Animal Genetics ,030217 neurology & neurosurgery ,Neuroscience - Abstract
Objective Substance P, encoded by the Tac1 gene, is involved in neurogenic inflammation and hyperalgesia via neurokinin 1 (NK1) receptor activation. Its non-neuronal counterpart, hemokinin-1, which is derived from the Tac4 gene, is also a potent NK1 agonist. Although hemokinin-1 has been described as a tachykinin of distinct origin and function compared to SP, its role in inflammatory and pain processes has not yet been elucidated in such detail. In this study, we analysed the involvement of tachykinins derived from the Tac1 and Tac4 genes, as well as the NK1 receptor in chronic arthritis of the mouse. Methods Complete Freund's Adjuvant was injected intraplantarly and into the tail of Tac1(-/-), Tac4(-/-), Tacr1(-/-) (NK1 receptor deficient) and Tac1(-/-/)Tac4(-/-) mice. Paw volume was measured by plethysmometry and mechanosensitivity using dynamic plantar aesthesiometry over a time period of 21 days. Semiquantitative histopathological scoring and ELISA measurement of IL-1β concentrations of the tibiotarsal joints were performed. Results Mechanical hyperalgesia was significantly reduced from day 11 in Tac4(-/-) and Tacr1(-/-) animals, while paw swelling was not altered in any strain. Inflammatory histopathological alterations (synovial swelling, leukocyte infiltration, cartilage destruction, bone damage) and IL-1β concentration in the joint homogenates were significantly smaller in Tac4(-/-) and Tac1(-/-/)Tac4(-/-) mice. Conclusions Hemokinin-1, but not substance P increases inflammation and hyperalgesia in the late phase of adjuvant-induced arthritis. While NK1 receptors mediate its antihyperalgesic actions, the involvement of another receptor in histopathological changes and IL-1β production is suggested.
- Published
- 2013
131. Absence of splenic latency in murine gammaherpesvirus 68-infected B cell-deficient mice
- Author
-
Anthony Nash, James P. Stewart, Edward J. Usherwood, Deborah J. Allen, and Kevin A. Robertson
- Subjects
CD4-Positive T-Lymphocytes ,viruses ,Lymphocyte ,Spleen ,Antibodies, Viral ,Virus ,Mice ,Gammaherpesvirinae ,Latent Virus ,Virology ,Virus latency ,medicine ,Animals ,Lung ,B cell ,B-Lymphocytes ,biology ,Murid herpesvirus 4 ,Antiviral antibody ,Herpesviridae Infections ,medicine.disease ,biology.organism_classification ,Virus Latency ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Splenomegaly ,Immunology - Abstract
Murine gammaherpesvirus 68 (MHV-68) is a natural pathogen of mice which causes an acute lung infection and establishes a latent infection in B lymphocytes. In this paper we describe the infection in transgenic B cell-deficient (muMT) mice, to determine whether a latent infection can be established in a mouse lacking circulating B lymphocytes. Little difference was observed in the acute lung infection, although there was a slight delay in virus clearance in the muMT mice. This indicates that antiviral antibody is of little importance in the resolution of the lung infection. Neither free nor latent virus could be detected in the spleen in the muMT mice. In addition, these mice did not develop MHV-68-induced splenomegaly. These data suggest that within the lymphoid compartment B lymphocytes are the sole reservoir for MHV-68 infection in vivo, confirming earlier work which identified B cells as the site of latent infection based on cell fractionation studies. In addition, our study shows that CD4-driven lymphocyte expansion leading to splenomegaly is dependent on the presence of MHV-68-infected B cells in the spleen. Although no free virus was detected (using conventional biological assays) in the lung after the resolution of the acute infection, MHV-68 genome was detected in the lungs of both control and muMT mice by PCR analysis. This suggests that cells in the lung may act as a reservoir of latent virus which is independent of the B lymphocyte infection.
- Published
- 1996
132. Murine Gammaherpesvirus-68 Encodes Homologues of Thymidine Kinase and Glycoprotein H: Sequence, Expression, and Characterization of Pyrimidine Kinase Activity
- Author
-
Stuart D Pepper, Arrand, James P. Stewart, and Mike Mackett
- Subjects
Genetics ,Base Sequence ,viruses ,Molecular Sequence Data ,Gene Expression ,Biology ,Thymidine Kinase ,Molecular biology ,Homology (biology) ,Mice ,Open Reading Frames ,Open reading frame ,Gammaherpesvirinae ,Viral Envelope Proteins ,Thymidine kinase ,Virology ,Animals ,RNA, Viral ,Coding region ,Amino Acid Sequence ,Northern blot ,ORFS ,Kinase activity ,Gene ,Glycoproteins - Abstract
We have sequenced a 4.5-kb fragment of DNA spanning the junction of the Bam HI D and E fragments of murine gammaherpesvirus-68 (MHV-68). This sequence was found to code for two major open reading frames (orfs) of 1934 and 2192 bp which showed significant homology to the thymidine kinase (TK) and glycoprotein H (gH) sequences of other gammaherpesviruses. Upstream from the TK gene another orf was found which showed amino acid sequence homology to the HSV1 UL24 gene. Analysis of the 1934-bp orf revealed the presence of all six of the recognized sites that are conserved between herpesvirus TKs although, uniquely among sequenced herpesvirus TK enzymes, MHV-68 lacks the consensus nucleotide binding site GXXGXGK, the second glycine being replaced by alanine. The MHV-68 TK has a predicted M r of 68,443, while the gH is predicted to have a M r of 82,890. Northern blot analysis showed an early TK message of 2.6 kb and a late gH-specific message of 2.5 kb. Both TK and gH probes detected a 4.3-kb late message, implying that this late message spans gH and TK. The TK coding sequence was expressed using an in vitro transcription translation system and was shown to encode functional TK activity.
- Published
- 1996
133. Antigenic and Sequence Variation in the C-Terminal Unique Domain of the Epstein-Barr Virus Nuclear Antigen EBNA-1
- Author
-
James P. Stewart, Mark N. Wrightham, Cliona M. Rooney, Clare E. Sample, Nusrat J. Janjua, John R. Arrand, and Stuart D Pepper
- Subjects
Herpesvirus 4, Human ,Recombinant Fusion Proteins ,T-Lymphocytes ,viruses ,Molecular Sequence Data ,Biology ,Antibodies, Viral ,medicine.disease_cause ,Major histocompatibility complex ,Epitope ,Virus ,Epitopes ,Antigen ,hemic and lymphatic diseases ,Virology ,HLA-A2 Antigen ,MHC class I ,medicine ,Humans ,Cytotoxic T cell ,Amino Acid Sequence ,Antigens, Viral ,Cell Line, Transformed ,B-Lymphocytes ,Base Sequence ,Antibodies, Monoclonal ,Genetic Variation ,Sequence Analysis, DNA ,Antigenic Variation ,Molecular biology ,Epstein–Barr virus ,DNA-Binding Proteins ,Epstein-Barr Virus Nuclear Antigens ,DNA, Viral ,biology.protein ,Antibody - Abstract
The Epstein-Barr virus (EBV) nuclear antigen EBNA-1 is essential for viral genome maintenance in vitro and may be the only EBV protein expressed by the majority of latently infected cells in vivo. EBNA-1 may therefore be critical to the evasion of host immunity which allows persistent infection. EBNA-1 includes a polymorphic internal repeat domain of unknown significance and unique regions which mediate all known functional activities and which have hitherto been assumed to be conserved between strains. Monoclonal antibodies were generated using a construct based on EBNA-1 of the prototype B95-8 strain, deleted for the repeat domain. These antibodies showed a limited profile of recognition of EBNA-1 in common laboratory EBV+ cell lines by immunoprecipitation and immunostaining. The observed antigenic heterogeneity also extended to spontaneously transformed B lymphoblastoid cell lines (LCLs) representing viral isolates circulating within US and UK populations. DNA fragments spanning the C-terminal unique domain of EBNA-1 from eleven spontaneous LCLs were amplified by polymerase chain reaction for sequencing, which directly demonstrated extensive and unexpected variability between diverse type 1 EBV isolates. The resulting polymorphism affects most of the putative MHC Class I binding epitopes which could be identified within this region using published sequence motifs, and influences MHC binding by variants of at least one such peptide in the processing mutant cell line T2. These findings could be related to the apparent lack of recognition of EBNA-1 by cytotoxic T lymphocytes.
- Published
- 1995
134. Distribution of Ovine Herpesvirus-2 Transcript and Protein in Naturally Infected Sheep
- Author
-
D.M. Amin, Anja Kipar, and James P. Stewart
- Subjects
General Veterinary ,Distribution (pharmacology) ,Biology ,Virology ,Ovine herpesvirus 2 ,Pathology and Forensic Medicine - Published
- 2016
135. BPIFA1: A Protein with an Important Role in the Inflammatory Response to Viral Infection in the Respiratory Tract
- Author
-
Colin D. Bingle, Nathifa Moyo, James P. Stewart, Mark Tompkins, Khondoker M. Akram, Gail E. Chapman, Tessa R. Walsh, Ralph A. Tripp, Lynne Bingle, and Gail Leeming
- Subjects
medicine.anatomical_structure ,General Veterinary ,business.industry ,Inflammatory response ,Immunology ,medicine ,A protein ,business ,Viral infection ,Pathology and Forensic Medicine ,Respiratory tract - Published
- 2016
136. Experimental infection of laboratory-bred bank voles (Myodes glareolus) with murid herpesvirus 4
- Author
-
David J. Hughes, James P. Stewart, Gail Leeming, Anja Kipar, and Jeffery T. Sample
- Subjects
Rhadinovirus ,Sinus histiocytosis ,Myodes glareolus ,Spleen ,Rodent Diseases ,Mice ,Virology ,medicine ,Animals ,Mice, Inbred BALB C ,biology ,Arvicolinae ,Murid herpesvirus 4 ,Alveolar epithelial cell ,Animal Structures ,General Medicine ,Herpesviridae Infections ,Models, Theoretical ,biology.organism_classification ,Bank vole ,Wood mouse ,Tumor Virus Infections ,medicine.anatomical_structure ,Genus Apodemus ,Murinae - Abstract
MuHV-4 is a natural pathogen of rodents of the genus Apodemus (e.g., wood mice, yellow-necked mice) and Myodes glareolus (bank voles). We report experimental MuHV-4 infection of bank voles in comparison with infection of A. sylvaticus (wood mice) and BALB/c mice. Like in wood mice, the level of productive replication in the lungs of bank voles was significantly lower than in BALB/c mice. In contrast to other hosts, however, the level of latent infection in the lung and spleen of bank voles was extremely low. These findings, together with those of previous studies, suggest that bank voles are an occasional and inefficient host for MuHV-4.
- Published
- 2012
137. Differential Expression of Viral and Human Interleukin-10 (IL-10) by Primary B Cell Tumors and B Cell Lines
- Author
-
John R. Arrand, James P. Stewart, Cliona M. Rooney, and Frederick G. Behm
- Subjects
Herpesvirus 4, Human ,Biopsy ,Molecular Sequence Data ,Naive B cell ,Biology ,medicine.disease_cause ,Models, Biological ,Cell Line ,Viral Proteins ,Virology ,Virus latency ,medicine ,Humans ,Autocrine signalling ,B cell ,B-Lymphocytes ,Base Sequence ,Cell Differentiation ,medicine.disease ,Burkitt Lymphoma ,Epstein–Barr virus ,Molecular biology ,Interleukin-10 ,Virus Latency ,medicine.anatomical_structure ,Lytic cycle ,Viral replication ,Cell culture ,DNA Probes - Abstract
Human and viral interleukin-10 (IL-10) possess growth factor activity for human B cells and may act as autocrine growth factors in B cell malignancies. To study this possibility we have measured viral (v) and human (h) IL-10 expression in EBV-positive and negative B lineage tumors and tumor cell lines. Previous studies demonstrated IL-10 expression in cell lines and now we describe the pattern of IL-10 expression in primary Burkitt's lymphoma (BL) tumor biopsies and in BL lines of defined phenotypes. vIL-10 was expressed during the lytic (productive) phase of EBV infection, but not during virus latency. Although hIL-10 was expressed in the majority of B cell lines, it was not expressed in two BL biopsy specimens. Expression of hIL-10 did not correlate with the presence of EBV, but was associated with the differentiation state of the B cell line. Thus, vIL-10 may enhance the persistence of B cells infected at sites of virus replication, and while hIL-10 may be a factor in the growth both in vivo and in vitro of some BLs and EBV-transformed B cells, it is not an absolute requirement.
- Published
- 1994
138. Chemokine binding protein M3 of murine gammaherpesvirus 68 modulates the host response to infection in a natural host
- Author
-
Elaine Bennett, Deborah Howarth, David J. Hughes, Jeffery T. Sample, R. Papoula-Pereira, Brian F. Flanagan, James P. Stewart, Joanne A. Cummerson, Gail Leeming, Anja Kipar, University of Zurich, and Früh, Klaus
- Subjects
Chemokine ,viruses ,2405 Parasitology ,Microbiology/Innate Immunity ,Mice ,0302 clinical medicine ,Cricetinae ,Virus latency ,Virology/Virulence Factors and Mechanisms ,Biology (General) ,Lung ,0303 health sciences ,2404 Microbiology ,virus diseases ,Herpesviridae Infections ,3. Good health ,Microbiology/Immunity to Infections ,Virus Latency ,Lymphatic system ,medicine.anatomical_structure ,Virology/Animal Models of Infection ,Chemokines ,Research Article ,Lymphoid Tissue ,QH301-705.5 ,Immunology ,10184 Institute of Veterinary Pathology ,Context (language use) ,Spleen ,Pathology/Immunology ,Bronchi ,Biology ,Virology/Immune Evasion ,Microbiology ,Cell Line ,03 medical and health sciences ,Viral Proteins ,Gammaherpesvirinae ,1311 Genetics ,Virology ,Immunology/Immunity to Infections ,Infectious Diseases/Viral Infections ,1312 Molecular Biology ,Genetics ,medicine ,Animals ,Molecular Biology ,030304 developmental biology ,2403 Immunology ,Virology/Persistence and Latency ,Germinal center ,RC581-607 ,medicine.disease ,Chemokine binding ,Viral replication ,2406 Virology ,biology.protein ,570 Life sciences ,biology ,Parasitology ,Murinae ,Immunologic diseases. Allergy ,030215 immunology - Abstract
Murine γ-herpesvirus 68 (MHV-68) infection of Mus musculus-derived strains of mice is an attractive model of γ-herpesvirus infection. Surprisingly, however, ablation of expression of MHV-68 M3, a secreted protein with broad chemokine-binding properties in vitro, has no discernable effect during experimental infection via the respiratory tract. Here we demonstrate that M3 indeed contributes significantly to MHV-68 infection, but only in the context of a natural host, the wood mouse (Apodemus sylvaticus). Specifically, M3 was essential for two features unique to the wood mouse: virus-dependent inducible bronchus-associated lymphoid tissue (iBALT) in the lung and highly organized secondary follicles in the spleen, both predominant sites of latency in these organs. Consequently, lack of M3 resulted in substantially reduced latency in the spleen and lung. In the absence of M3, splenic germinal centers appeared as previously described for MHV-68-infected laboratory strains of mice, further evidence that M3 is not fully functional in the established model host. Finally, analyses of M3's influence on chemokine and cytokine levels within the lungs of infected wood mice were consistent with the known chemokine-binding profile of M3, and revealed additional influences that provide further insight into its role in MHV-68 biology., Author Summary Infection of inbred strains of laboratory mice (Mus musculus) with the rodent γ-herpesvirus MHV-68 continues to be developed as an attractive experimental model of γ-herpesvirus infection. In this regard, the MHV-68 protein M3 has been shown to selectively bind and inhibit chemokines involved in the antiviral immune response, a property expected to contribute significantly to virus infection and host colonization. However, inactivation of the M3 gene has no discernable consequence on infection in this animal host. Prompted by recent evidence that natural hosts of MHV-68 are members of the genus Apodemus, and that MHV-68 infection in laboratory-bred wood mice (Apodemus sylvaticus) differs significantly from that which has been described in standard strains of laboratory mice, we addressed whether M3 functions in a host-specific manner. Indeed, we find that M3 is responsible for host-specific differences observed for MHV-68 infection, that its influence on infection within wood mice is consistent with its chemokine-binding properties, and that in its absence, persistent latent infection - a hallmark of herpesvirus infections - is attenuated. This highlights the importance of host selection when investigating specific roles of pathogenesis-related viral genes, and advances our understanding of this model and its potential application to human γ-herpesvirus infections.
- Published
- 2011
139. Ganciclovir Antiviral Therapy in Advanced Idiopathic Pulmonary Fibrosis: An Open Pilot Study
- Author
-
S.S. Lok, Jim J. Egan, Ashley Woodcock, Huzaifa Adamali, and James P. Stewart
- Subjects
Pulmonary and Respiratory Medicine ,Ganciclovir ,lcsh:RC705-779 ,medicine.medical_specialty ,Vital capacity ,Article Subject ,business.industry ,Disease progression ,Antiviral therapy ,General Medicine ,lcsh:Diseases of the respiratory system ,medicine.disease ,Gastroenterology ,Surgery ,Pathogenesis ,Idiopathic pulmonary fibrosis ,FEV1/FVC ratio ,Internal medicine ,medicine ,Prednisolone ,Clinical Study ,business ,medicine.drug - Abstract
Hypothesis. Repeated epithelial cell injury secondary to viruses such as Epstein Barr and subsequent dysfunctional repair may be central to the pathogenesis of IPF. In this observational study, we evaluated whether a combination of standard and anti-viral therapy might have an impact on disease progression.Methods. Advanced IPF patients who failed standard therapy and had serological evidence of previous EBV, received ganciclovir (iv) at 5 mg/kg twice daily. Forced vital capacity (FVC), shuttle walk test, DTPA scan and prednisolone dose were measured before and 8 weeks post-treatment.Results. Fourteen patients were included. After ganciclovir, eight patients showed improvement in FVC and six deteriorated. The median reduction of prednisolone dose was 7.5 mg (44%). Nine patients were classified “responders” of whom four showed an improvement in all four criteria, while three of the five “non-responders” showed no response in any of the criteria. Responders showed reduction in prednisolone dosage (P=.02) and improved DTPA clearance (P=.001).Conclusion. This audit outcome suggests that 2-week course of ganciclovir (iv) may attenuate disease progression in a subgroup of advanced IPF patients. These observations do not suggest that anti-viral treatment is a substitute for the standard care, however, suggests the need to explore the efficacy of ganciclovir as adjunctive therapy in IPF.
- Published
- 2011
- Full Text
- View/download PDF
140. The Epstein-Barr Virus Candidate Vaccine Antigen gp340/220 Is Highly Conserved between Virus Types A and B
- Author
-
Michael Mackett, Stuart D Pepper, Janice F. Lees, James P. Stewart, Jane E. Arrand, and John R. Arrand
- Subjects
Herpesvirus 4, Human ,Antigenicity ,Molecular Sequence Data ,Fluorescent Antibody Technique ,Biology ,medicine.disease_cause ,Polymerase Chain Reaction ,Epitope ,Virus ,Cell Line ,Conserved sequence ,Viral Matrix Proteins ,Antigen ,hemic and lymphatic diseases ,Virology ,medicine ,Humans ,Coding region ,Amino Acid Sequence ,Antigens, Viral ,Gene ,Conserved Sequence ,Genetics ,Base Sequence ,Sequence Homology, Amino Acid ,Antibodies, Monoclonal ,Viral Vaccines ,Epstein–Barr virus ,DNA, Viral - Abstract
Anti-Epstein-Barr Virus (EBV) vaccines are being developed which are based on the gp340/220 membrane antigen (MA) gene products from the B95-8 strain. Some proteins are known to be immunologically quite different between type-A (1) and type-B (2) strains of EBV and therefore from a vaccine point of view it was critical to evaluate the degree of conservation of gp340/220. The complete MA coding sequence was determined for two B-type viruses, AG876 and P3HR-1, for comparison with the A-type B95-8. A variable region within MA was sequenced from several other strains. In addition the other open reading frames within the MA-containing BamHI-L fragment of AG876 were sequenced and compared. The results show that there is a high degree of homology between all strains examined. Although some differences were found within the MA coding sequence the only major site of variation was within the repeat region and no consistent A/B changes were found. Monoclonal antibodies generated against A-type MA and representing six epitope groups along the length of the gp340 molecule were found to recognize B-type gp340, thereby demonstrating functional homology. We conclude that, as a vaccine antigen, B95-8gp340/220 should be equally effective against both types of EBV.
- Published
- 1993
141. Expression of the Epstein-Barr virus latent membrane protein in nasopharyngeal carcinoma biopsy specimens
- Author
-
John R. Arrand and James P. Stewart
- Subjects
China ,Herpesvirus 4, Human ,medicine.drug_class ,Biopsy ,viruses ,Fluorescent Antibody Technique ,Biology ,medicine.disease_cause ,Monoclonal antibody ,Virus ,Herpesviridae ,Pathology and Forensic Medicine ,Viral Matrix Proteins ,Antigen ,hemic and lymphatic diseases ,otorhinolaryngologic diseases ,medicine ,Humans ,Gammaherpesvirinae ,Antigens, Viral ,medicine.diagnostic_test ,Antibodies, Monoclonal ,Nasopharyngeal Neoplasms ,medicine.disease ,biology.organism_classification ,Epstein–Barr virus ,Virology ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,stomatognathic diseases ,Cell Transformation, Neoplastic ,Epstein-Barr Virus Nuclear Antigens ,Nasopharyngeal carcinoma ,Cancer research - Abstract
It has been known for some time that the Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC). The tumor cells are known to harbor EBV in a latent state. Latently-infected B cells that have become growth transformed by EBV in vitro express some 10 antigens, two of which (Epstein-Barr nuclear antigen 2 [EBNA2] and the latent membrane protein [LMP]) are associated with cellular transformation. We evaluated the expression of these two EBV antigens in NPC by probing tissue sections with monoclonal antibodies. We found that EBNA2 was not expressed and that LMP was expressed in seven of nine biopsy specimens. It is therefore postulated that either there are subsets of NPC or that LMP may be involved only in certain stages of tumor formation.
- Published
- 1993
142. Erratum to: Ljungan virus is endemic in rodents in the UK
- Author
-
Anne-Marie Salisbury, Winifred Dove, Bo Niklasson, James P. Stewart, and Michael Begon
- Subjects
medicine.medical_specialty ,Medical microbiology ,Ljungan virus ,Virology ,medicine ,General Medicine ,Biology - Published
- 2014
143. A Gammaherpesvirus Complement Regulatory Protein Promotes Initiation of Infection by Activation of Protein Kinase Akt/PKB
- Author
-
Heiko Adler, James P. Stewart, Barbara Adler, Beatrix Steer, and Stipan Jonjić
- Subjects
Science ,viruses ,Biology ,Virus ,Mice ,Open Reading Frames ,Viral Proteins ,Gammaherpesvirinae ,protein kinase Akt/PKB ,ORF4 ,gammaherpesvirus RCA protein ,Virology ,Animals ,Insulin ,Virology/Virulence Factors and Mechanisms ,Phosphorylation ,Protein kinase A ,Protein kinase B ,Glycosaminoglycan binding ,Multidisciplinary ,Reverse Transcriptase Polymerase Chain Reaction ,Kinase ,Genetic Complementation Test ,BIOMEDICINE AND HEALTHCARE. Basic Medical Sciences ,immune response ,protein VCP ,Virology/Host Invasion and Cell Entry ,Flow Cytometry ,Molecular biology ,Complement system ,Kinetics ,Virology/Viral Replication and Gene Regulation ,Microscopy, Fluorescence ,NIH 3T3 Cells ,Medicine ,Ribonucleosides ,Signal transduction ,BIOMEDICINA I ZDRAVSTVO. Temeljne medicinske znanosti ,Proto-Oncogene Proteins c-akt ,Research Article ,Signal Transduction - Abstract
BackgroundViruses have evolved to evade the host's complement system. The open reading frames 4 (ORF4) of gammaherpesviruses encode homologs of regulators of complement activation (RCA) proteins, which inhibit complement activation at the level of C3 and C4 deposition. Besides complement regulation, these proteins are involved in heparan sulfate and glycosaminoglycan binding, and in case of MHV-68, also in viral DNA synthesis in macrophages.Methodology/principal findingsHere, we made use of MHV-68 to study the role of ORF4 during infection of fibroblasts. While attachment and penetration of virions lacking the RCA protein were not affected, we observed a delayed delivery of the viral genome to the nucleus of infected cells. Analysis of the phosphorylation status of a variety of kinases revealed a significant reduction in phosphorylation of the protein kinase Akt in cells infected with ORF4 mutant virus, when compared to cells infected with wt virus. Consistent with a role of Akt activation in initial stages of infection, inhibition of Akt signaling in wt virus infected cells resulted in a phenotype resembling the phenotype of the ORF4 mutant virus, and activation of Akt by addition of insulin partially reversed the phenotype of the ORF4 mutant virus. Importantly, the homologous ORF4 of KSHV was able to rescue the phenotype of the MHV-68 ORF4 mutant, indicating that ORF4 is functionally conserved and that ORF4 of KSHV might have a similar function in infection initiation.Conclusions/significanceIn summary, our studies demonstrate that ORF4 contributes to efficient infection by activation of the protein kinase Akt and thus reveal a novel function of a gammaherpesvirus RCA protein.
- Published
- 2010
144. The interleukin-10 homolog encoded by Eepstein-Barr virus enhances the reactivation of virus-specific cytotoxic T cell and HLA-unrestricted killer cell responses
- Author
-
Cliona M. Rooney and James P. Stewart
- Subjects
Cytotoxicity, Immunologic ,Herpesvirus 4, Human ,Biology ,medicine.disease_cause ,Virus ,Cell Line ,Natural killer cell ,Immune system ,HLA Antigens ,hemic and lymphatic diseases ,Virology ,medicine ,Humans ,Cytotoxic T cell ,B cell ,B-Lymphocytes ,Sequence Homology, Amino Acid ,ZAP70 ,Natural killer T cell ,Epstein–Barr virus ,Recombinant Proteins ,Interleukin-10 ,Killer Cells, Natural ,medicine.anatomical_structure ,Interleukin-2 ,T-Lymphocytes, Cytotoxic - Abstract
We determined what influence the Epstein-Barr virus (EBV)-encoded homologue of IL-10 (viral or vIL-10) had on immune responses important for the control of EBV infection. We produced recombinant vIL-10 in a B cell line. A 17-kDa recombinant protein was secreted and had the same molecular weight as vIL-10 secreted by EBV-infected B cells. Functional activity of recombinant vIL-10 was shown by the inhibition of interferon-gamma production by activated leukocytes. Cytotoxic T cells and HLA-unrestricted activated killer cells are both important arms of the immune response to EBV. vIL-10, either expressed by B cell stimulators or added exogenously, enhanced the in vitro reactivation of allo- and EBV-specific cytotoxic T cells. vIL-10 also enhanced the activation of HLA-unrestricted killer cells by EBV-transformed B cells. In contrast, the interleukin-2-mediated activation of these killers was unaffected. Since vIL-10 is expressed during the lytic cycle of EBV, we conclude that the expression of vIL-10 may enhance immune responses to EBV-infected cells during periods of virus replication in vivo. In this way, the virus may limit its own replication and maintain the apathogenic virus carrier state that is characteristic of EBV.
- Published
- 1992
145. Characterization of a novel wood mouse virus related to murid herpesvirus 4
- Author
-
Claude Chastel, Charles Cunningham, Marie Adele Rajandream, Anja Kipar, Malcolm J. Bennett, Michael A. Quail, Rory J. Bowden, David J. Hughes, Jeffery T. Sample, Andrew J. Davison, Stacey Efstathiou, James P. Stewart, Mandy Sanders, Bart Barrell, and Steven G. Milligan
- Subjects
Rhadinovirus ,Field vole ,viruses ,Molecular Sequence Data ,Genome, Viral ,Genome ,Virus ,Viral Matrix Proteins ,03 medical and health sciences ,Gammaherpesvirinae ,Virology ,Animals ,Amino Acid Sequence ,Gene ,030304 developmental biology ,Genetics ,0303 health sciences ,biology ,Base Sequence ,030306 microbiology ,Arvicolinae ,Animal ,Murid herpesvirus 4 ,Nucleic acid sequence ,biology.organism_classification ,3. Good health ,DNA, Viral ,Murinae - Abstract
Two novel gammaherpesviruses were isolated, one from a field vole (Microtus agrestis) and the other from wood mice (Apodemus sylvaticus). The genome of the latter, designated wood mouse herpesvirus (WMHV), was completely sequenced. WMHV had the same genome structure and predicted gene content as murid herpesvirus 4 (MuHV4; murine gammaherpesvirus 68). Overall nucleotide sequence identity between WMHV and MuHV4 was 85 % and most of the 10 kb region at the left end of the unique region was particularly highly conserved, especially the viral tRNA-like sequences and the coding regions of genes M1 and M4. The partial sequence (71 913 bp) of another gammaherpesvirus, Brest herpesvirus (BRHV), which was isolated ostensibly from a white-toothed shrew (Crocidura russula), was also determined. The BRHV sequence was 99.2 % identical to the corresponding portion of the WMHV genome. Thus, WMHV and BRHV appeared to be strains of a new virus species. Biological characterization of WMHV indicated that it grew with similar kinetics to MuHV4 in cell culture. The pathogenesis of WMHV in wood mice was also extremely similar to that of MuHV4, except for the absence of inducible bronchus-associated lymphoid tissue at day 14 post-infection and a higher load of latently infected cells at 21 days post-infection.
- Published
- 2009
146. Ovine herpesvirus 2 structural proteins in epithelial cells and M-cells of the appendix in rabbits with malignant catarrhal fever
- Author
-
Kurt Tobler, Claudia S. Meier-Trummer, James P. Stewart, David M. Haig, Daniel L. Glauser, Jane Hart, Mathias Ackermann, Iris Campbell, Felix Ehrensperger, Monika Hilbe, University of Zurich, and Ackermann, M
- Subjects
3400 General Veterinary ,10184 Institute of Veterinary Pathology ,In situ hybridization ,Biology ,Appendix ,medicine.disease_cause ,Microbiology ,Herpesviridae ,Virus ,Antigen ,medicine ,Animals ,General Veterinary ,2404 Microbiology ,Epithelial Cells ,General Medicine ,Viral tegument ,Herpesviridae Infections ,Virology ,Molecular biology ,In vitro ,Viral replication ,Capsid ,Malignant Catarrh ,570 Life sciences ,biology ,Rabbits ,10244 Institute of Virology - Abstract
Sheep-associated malignant catarrhal fever (MCF), caused by Ovine herpesvirus 2 (OvHV-2), is a usually fatal disease of various ruminants and swine. A system for propagation of OvHV-2 in vitro has not yet been identified, although persistently infected cells have been derived from diseased animals and used to establish an animal model in rabbits. OvHV-2 structural proteins have not been detected in diseased animals and the pathogenesis of OvHV-2 infection is poorly understood. Recently, the genomic sequence of OvHV-2 has been determined, which allowed to predict the amino acid sequences of putative OvHV-2 structural proteins. Based on those predictions, we have generated antisera against two putative structural proteins (ORF43 and ORF63) of OvHV-2 in order to detect sites of active virus replication in experimentally OvHV-2-infected rabbits with signs of MCF. Although histological lesions typical of MCF were detected in multiple tissues, those sera detected viral capsid and tegument antigens exclusively in the appendix but not in other tissues of rabbits with MCF. More specifically, those viral proteins were detected in epithelial cells as well as in M-cells. However, in situ hybridization revealed that ORF63 mRNA was present in epithelial cells of infected rabbits but not in M-cells. Our data suggest that active OvHV-2 replication takes place in certain tissues of animals with MCF and that M-cells may play a role in the athogenesis of MCF.
- Published
- 2009
147. The interaction of the gammaherpesvirus 68 orf73 protein with cellular BET proteins affects the activation of cell cycle promoters
- Author
-
Thomas Christalla, Ronald Frank, James P. Stewart, Thomas F. Schulz, Daniel Pliquet, Matthias Ottinger, and Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
- Subjects
BRD4 ,Protein family ,Transcription, Genetic ,viruses ,Immunology ,Molecular Sequence Data ,Biology ,Protein Serine-Threonine Kinases ,Microbiology ,Retinoblastoma Protein ,Cell Line ,Retinoblastoma-like protein 1 ,Mice ,Viral Proteins ,Cyclin D2 ,Virology ,Animals ,Humans ,Amino Acid Sequence ,Promoter Regions, Genetic ,Herpesviridae ,Cell Nucleus ,Binding Sites ,General transcription factor ,Cell Cycle ,virus diseases ,TAF7 ,Protein-Serine-Threonine Kinases ,Molecular biology ,Chromatin ,Bromodomain ,Virus-Cell Interactions ,GATAD2B ,Insect Science ,Mutation ,Sequence Alignment ,Protein Binding - Abstract
Many gammaherpesviruses promote lymphocyte proliferation and can persist in the lymphocyte compartment as well as in a number of other cell types, including endothelial and epithelial cells and fibroblasts (16, 17, 19, 56, 59). To ensure episome maintenance in these dividing cells, gammaherpesviruses express proteins that facilitate the replication of latent viral episomes and ensure the segregation of these viral genomes into progeny cells during cell division. The Epstein-Barr virus (EBV) EBNA-1 protein fulfills these essential functions (49, 62), as do the proteins encoded by open reading frame 73 (ORF73) of the gamma-2 herpesviruses (rhadinoviruses) Kaposi's sarcoma-associated herpesvirus (KSHV), herpesvirus saimiri (HVS), and murine gammaherpesvirus 68 (MHV-68) (4, 20, 28, 29, 43). The orf73 protein of KSHV, latency associated nuclear antigen 1 (LANA-1), is well characterized and has multiple functions. LANA-1 mediates replication and episome maintenance, acts as a transcriptional repressor and activator, and deregulates the cell division cycle (3, 24, 50). Some of these functions are mediated via the interaction of LANA-1 with a number of cellular proteins, including p53 (23), the retinoblastoma protein (48), MeCP2 (35), mSin3 (36), and multiple members of the BET (bromodomain and extra terminal domain) family of proteins (45, 47, 65). The MHV-68 orf73 protein, the KSHV LANA-1 homolog, is critical for the establishment and maintenance of a latent infection in mice (20, 43). MHV-68 orf73 is expressed in latently infected cells as well as during lytic infection (2, 15, 19, 51). The molecular details of how the MHV-68 protein functions are still largely unexplored. BET proteins interact via their bromodomains with acetylated histones (13, 30, 33) and are highly conserved, with members in plants, yeast, Drosophila melanogaster, and up to mammals (18). An additional characteristic feature of BET proteins is the highly conserved extra terminal (ET) domain (see Fig. Fig.1),1), which serves as a protein-protein interaction module in Brd2, Brd3, and Brd4 with KSHV LANA-1 (45, 47, 65) and between the yeast (Saccharomyces cerevisiae) BET protein Bdf1 and the TAF7 subunit of the general transcription factor TFIID (39). Mammalian BET proteins are encoded by four genes, Brd2, Brd3, Brd4, and Brd6, out of which Brd2, also called RING3, and Brd4 are the best characterized. Brd2 is a transcriptional regulator that plays a role in cell cycle regulation (11, 12, 33, 55). Brd2 overexpression in B lymphocytes in vivo in mice has been shown to induce lymphoma (26). Brd4 interacts with pTEFb (transcriptional elongation factor b) and thereby promotes RNA polymerase II (Pol II)-dependent transcriptional elongation (7, 32, 61). Brd4 overexpression and depletion both result in deregulated cell cycle progression (14, 38, 45). Recently, Brd4 has been shown to play an important role in the G1/S transition through its ability to stimulate transcription of G1/S-specific genes, among them, cyclin D1 and cyclin D2 (42). Furthermore, the C-terminal domain of the long isoform of Brd4 (Brd4L) (amino acids [aa] 1 to 1362) serves as a chromatin tether for different papillomaviruses via its interaction with their E2 proteins (1, 6, 63, 64). Brd4 is critical for papillomavirus E2 transcriptional activation (31, 40, 54) and may play a role in its transcriptional repression function (53, 60). The same C-terminal domain of Brd4L has recently been shown to interact with the transcriptional elongation factor pTEF and to inhibit human immunodeficiency virus type 1 (HIV-1) Tat-mediated, pTEF-dependent transcription (7). We have shown previously that KSHV LANA-1 interacts with Brd2 and Brd4 via their conserved ET domains (45, 57, 58). This interaction contributes to the association of LANA-1 with cellular heterochromatin and modulates the transcriptional activator role of the short isoform of Brd4 (Brd4S) (aa 1 to 722), which lacks the pTEFb interaction domain and by itself activates the cyclin E promoter (7, 45, 57, 58). FIG. 1. The MHV-68 orf73 protein interacts with the carboxy termini of the cellular BET proteins Brd2/RING3, Brd4S, and Brd3/ORFX. (A) Schematic depiction of the human BET proteins. BR1, bromodomain 1; BR2, bromodomain 2. (B) High degree of sequence conservation ... In this study we show that Brd2, Brd3, and Brd4 also interact with the MHV-68 orf73 protein. By identifying and mutating a binding site for Brd4 and Brd2 in the MHV-68 orf73 protein, we show that the orf73/BET interaction is crucial for the ability to activate the cyclin D1, D2, and E promoters. The results pinpoint the binding site for two BET proteins in a rhadinoviral orf73 protein and indicate that, similarly to papillomavirus E2, a rhadinoviral orf73 protein utilizes a member of the BET protein family to exert its transcriptional activation function.
- Published
- 2009
148. Induction of Tachykinin Production in Airway Epithelia in Response to Viral Infection
- Author
-
Helen Cox, Catherine Payne, Sylvia A. Vasiliou, John P. Quinn, James P. Stewart, and Anja Kipar
- Subjects
Chemokine ,Respiratory System ,lcsh:Medicine ,Substance P ,Inflammation ,Mice, Transgenic ,Biology ,Epithelium ,Proinflammatory cytokine ,Immunoenzyme Techniques ,chemistry.chemical_compound ,Mice ,Immune system ,Gammaherpesvirinae ,Immunology/Immunity to Infections ,Tachykinins ,medicine ,Animals ,heterocyclic compounds ,Protein Precursors ,lcsh:Science ,Medicine(all) ,Neurogenic inflammation ,Mice, Inbred BALB C ,Multidisciplinary ,Lung ,Agricultural and Biological Sciences(all) ,Biochemistry, Genetics and Molecular Biology(all) ,Neuroscience/Sensory Systems ,Macrophages ,lcsh:R ,Herpesviridae Infections ,beta-Galactosidase ,Peptide Fragments ,medicine.anatomical_structure ,chemistry ,Immunology ,Immunology/Immune Response ,biology.protein ,Virology/Animal Models of Infection ,lcsh:Q ,medicine.symptom ,Respiratory tract ,Research Article - Abstract
Background: The tachykinins are implicated in neurogenic inflammation and the neuropeptide substance P in particular has been shown to be a proinflammatory mediator. A role for the tachykinins in host response to lung challenge has been previously demonstrated but has been focused predominantly on the release of the tachykinins from nerves innervating the lung. We have previously demonstrated the most dramatic phenotype described for the substance P encoding gene preprotachykinin-A (PPT-A) to date in controlling the host immune response to the murine gammaherpesvirus 68, in the lung.Methodology/ Principal Findings: In this study we have utilised transgenic mice engineered to co-ordinately express the beta-galactosidase marker gene along with PPT-A to facilitate the tracking of PPT-A expression. Using a combination of these mice and conventional immunohistology we now demonstrate that PPT-A gene expression and substance P peptide are induced in cells of the respiratory tract including tracheal, bronchiolar and alveolar epithelial cells and macrophages after viral infection. This induction was observed 24h post infection, prior to observable inflammation and the expression of pro-inflammatory chemokines in this model. Induced expression of the PPT-A gene and peptide persisted in the lower respiratory tract through day 7 post infection.Conclusions/ Significance: Non-neuronal PPT-A expression early after infection may have important clinical implications for the progression or management of lung disease or infection aside from the well characterised later involvement of the tachykinins during the inflammatory response.
- Published
- 2008
149. Serological survey of virus infection among wild house mice (Mus domesticus) in the UK
- Author
-
Jane L. Hurst, James P. Stewart, Stuart D. Becker, and Malcolm J. Bennett
- Subjects
education.field_of_study ,General Veterinary ,biology ,viruses ,Murid herpesvirus 4 ,Population ,virus diseases ,Animals, Wild ,biology.organism_classification ,Antibodies, Viral ,Virology ,Sendai virus ,Virus ,United Kingdom ,Rodent Diseases ,Mice ,Mouse hepatitis virus ,Seroepidemiologic Studies ,Virus Diseases ,Animals ,Animal Science and Zoology ,Orthopoxvirus ,House mice ,education ,Minute virus of mice - Abstract
The serological prevalence of 13 murine viruses was surveyed among 103 wild-caught and 51 captive-bred house mice ( Mus domesticus), originating from several trapping locations in northwest England, using blood samples obtained during routine health screening of an established wild mouse colony. A high proportion of recently caught wild mice were seropositive for mouse hepatitis virus (86%), mouse cytomegalovirus (79%), mouse thymic virus (78%), mouse adenovirus (68%), mouse parvovirus (59%) and minute virus of mice (41%). Seroprevalences of lymphocytic choriomeningitis virus (LCMV), orthopoxvirus, reovirus-3 and murid herpesvirus 4 (MuHV-4, also called murine γ-herpesvirus [MHV-68]) were low (3–13%), and no animals were seropositive to Sendai virus, pneumonia virus or polyomavirus. Seroprevalence in wild-caught animals that had been in captivity for over six months was generally consistent with the range found in recently caught wild animals, while seroprevalence was generally much lower in captive-bred mice despite no attempt to prevent viral spread. A notable exception to this was LCMV, which appeared to have spread efficiently through the captive population (both captive-bred and wild-caught animals). Given the known viral life cycles in laboratory mice, it appears that viral persistence in the host was an important contributing factor in the spread of infection in captivity.
- Published
- 2007
150. Complete sequence and analysis of the ovine herpesvirus 2 genome
- Author
-
James P. Stewart, David M. Haig, Jane Hart, Gamini Jayawardane, Mathias Ackermann, Hugh W. Reid, George C. Russell, University of Zurich, and Stewart, J P
- Subjects
Rhadinovirus ,Genes, Viral ,Sequence analysis ,Molecular Sequence Data ,Genome, Viral ,Biology ,Genome ,Homology (biology) ,Cell Line ,Open Reading Frames ,Complete sequence ,Exon ,Virology ,Animals ,ORFS ,Gene ,Genetics ,Sheep ,Sequence Analysis, DNA ,Open reading frame ,DNA, Viral ,2406 Virology ,570 Life sciences ,biology ,Cattle ,10244 Institute of Virology - Abstract
Ovine herpesvirus 2 (OvHV-2) is endemic in sheep populations worldwide and causes malignant catarrhal fever (MCF), a lymphoproliferative disease, in cattle, bison and deer. OvHV-2 has been placed in the gammaherpesvirus subfamily and is related closely to Alcelaphine herpesvirus 1 (AlHV-1). Here, the cloning, sequencing and analysis of the complete OvHV-2 genome derived from a lymphoblastoid cell line from an affected cow (BJ1035) are reported. The unique portion of the genome consists of 130 930 bp, with a mean G+C content of 52 mol%. The unique DNA is flanked by multiple copies of terminal repeat elements 4205 bp in length, with a mean G+C content of 72 mol%. Analysis revealed 73 open reading frames (ORFs), the majority (62) of which showed homology to other gammaherpesvirus genes. A further subset of nine ORFs is shared with only the related AlHV-1. Three ORFs are entirely unique to OvHV-2, including a spliced homologue of cellular interleukin-10 that retains the exon structure of the cellular gene. The sequence of OvHV-2 is a critical first step in the study of the pathogenesis and treatment of MCF.
- Published
- 2007
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.