bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Recombinant Human Monoclonal Antibody to Human Metapneumovirus Fusion Protein That Neutralizes Virus In Vitro and Is Effective Therapeutically In Vivo
Authors: Williams John V. , Chen Zhifeng , Cseke Gabriella , Wright David W. , Keefer Christopher J. , Tollefson Sharon J. , Hessell Ann , Podsiad Amy , Shepherd Bryan E. , Sanna Pietro Paolo , Burton Dennis R. , Crowe James E. , Williamson R. Anthony 
Published: May 2007

ABSTRACTHuman metapneumovirus (hMPV) is a recently discovered paramyxovirus that is a major cause of lower-respiratory-tract disease. hMPV is associated with more severe disease in infants and persons with underlying medical conditions. Animal studies have shown that the hMPV fusion (F) protein alone is capable of inducing protective immunity. Here, we report the use of phage display technology to generate a fully human monoclonal antibody fragment (Fab) with biological activity against hMPV. Phage antibody libraries prepared from human donor tissues were selected against recombinant hMPV F protein with multiple rounds of panning. Recombinant Fabs then were expressed in bacteria, and supernatants were screened by enzyme-linked immunosorbent assay and immunofluorescent assays. A number of Fabs that bound to hMPV F were isolated, and several of these exhibited neutralizing activity in vitro. Fab DS7 neutralized the parent strain of hMPV with a 60% plaque reduction activity of 1.1 μg/ml and bound to hMPV F with an affinity of 9.8 ×10−10M, as measured by surface plasmon resonance. To test the in vivo activity of Fab DS7, groups of cotton rats were infected with hMPV and given Fab intranasally 3 days after infection. Nasal turbinates and lungs were harvested on day 4 postinfection and virus titers determined. Animals treated with Fab DS7 exhibited a >1,500-fold reduction in viral titer in the lungs, with a modest 4-fold reduction in the nasal tissues. There was a dose-response relationship between the dose of DS7 and virus titer. Human Fab DS7 may have prophylactic or therapeutic potential against severe hMPV infection.
A Functional Heat Shock Protein 90 Chaperone Is Essential for Efficient Flock House Virus RNA Polymerase Synthesis in
Drosophila
Cells
Authors: Castorena Kathryn M. , Weeks Spencer A. , Stapleford Kenneth A. , Cadwallader Amy M. , Miller David J. 
Published: May 2007

ABSTRACT

The molecular chaperone heat shock protein 90 (Hsp90) is involved in multiple cellular processes including protein maturation, complex assembly and disassembly, and intracellular transport. We have recently shown that a disruption of Hsp90 activity in cultured
Drosophila melanogaster
cells suppresses Flock House virus (FHV) replication and the accumulation of protein A, the FHV RNA-dependent RNA polymerase. In the present study, we investigated whether the defect in FHV RNA polymerase accumulation induced by Hsp90 suppression was secondary to an effect on protein A synthesis, degradation, or intracellular membrane association. Treatment with the Hsp90-specific inhibitor geldanamycin selectively reduced FHV RNA polymerase synthesis by 80% in
Drosophila
S2 cells stably transfected with an inducible protein A expression plasmid. The suppressive effect of geldanamycin on protein A synthesis was not attenuated by proteasome inhibition, nor was it sensitive to changes in either the mRNA untranslated regions or protein A intracellular membrane localization. Furthermore, geldanamycin did not promote premature protein A degradation, nor did it alter the extremely rapid kinetics of protein A membrane association. These results identify a novel role for Hsp90 in facilitating viral RNA polymerase synthesis in
Drosophila
cells and suggest that FHV subverts normal cellular pathways to assemble functional replication complexes.
Chicken Heat Shock Protein 90 Is a Component of the Putative Cellular Receptor Complex of Infectious Bursal Disease Virus
Authors: Lin Ta-Wei , Lo Chi-Wen , Lai Su-Yuan , Fan Ruey-Jane , Lo Chao-Jung , Chou Yu-mei , Thiruvengadam Rekha , Wang Andrew H.-J. , Wang Min-Ying 
Published: May 2007

ABSTRACT

Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. The capsid protein VP2 of IBDV plays an important role in virus binding and cell recognition. VP2 forms a subviral particle (SVP) with immunogenicity similar to that of the IBDV capsid. In the present study, we first showed that SVP could inhibit IBDV infection to an IBDV-susceptible cell line, DF-1 cells, in a dose-dependent manner. Second, the localizations of the SVP on the surface of DF-1 cells were confirmed by fluorescence microscopy, and the specific binding of the SVP to DF-1 cells occurred in a dose-dependent manner. Furthermore, the attachment of SVP to DF-1 cells was inhibited by an SVP-induced neutralizing monoclonal antibody against IBDV but not by denatured-VP2-induced polyclonal antibodies. Third, the cellular factors in DF-1 cells involved in the attachment of SVP were purified by affinity chromatography using SVP bound on the immobilized Ni
2+
ions. A dominant factor was identified as being chicken heat shock protein 90 (Hsp90) (cHsp90) by mass spectrometry. Results of biotinylation experiments and indirect fluorescence assays indicated that cHsp90 is located on the surface of DF-1 cells. Virus overlay protein binding assays and far-Western assays also concluded that cHsp90 interacts with IBDV and SVP, respectively. Finally, both Hsp90 and anti-Hsp90 can inhibit the infection of DF-1 cells by IBDV. Taken together, for the first time, our results suggest that cHsp90 is part of the putative cellular receptor complex essential for IBDV entry into DF-1 cells.
Major Neutralizing Sites on Vaccinia Virus Glycoprotein B5 Are Exposed Differently on Variola Virus Ortholog B6
Authors: Aldaz-Carroll Lydia , Xiao Yuhong , Whitbeck J. Charles , de Leon Manuel Ponce , Lou Huan , Kim Mikyung , Yu Jessica , Reinherz Ellis L. , Isaacs Stuart N. , Eisenberg Roselyn J. , Cohen Gary H. 
Published: May 2007

ABSTRACT
Immunization against smallpox (variola virus) with Dryvax, a live vaccinia virus (VV), was effective, but now safety is a major concern. To overcome this issue, subunit vaccines composed of VV envelope proteins from both forms of infectious virions, including the extracellular enveloped virion (EV) protein B5, are being developed. However, since B5 has 23 amino acid differences compared with its B6 variola virus homologue, B6 might be a better choice for such a strategy. Therefore, we compared the properties of both proteins using a panel of monoclonal antibodies (MAbs) to B5 that we had previously characterized and grouped according to structural and functional properties. The B6 gene was obtained from the Centers for Disease Control and Prevention, and the ectodomain was cloned and expressed in baculovirus as previously done with B5, allowing us to compare the antigenic properties of the proteins. Polyclonal antibodies to B5 or B6 cross-reacted with the heterologous protein, and 16 of 26 anti-B5 MAbs cross-reacted with B6. Importantly, 10 anti-B5 MAbs did not cross-react with B6. Of these, three have important anti-VV biologic properties, including their ability to neutralize EV infectivity and block comet formation. Here, we found that one of these three MAbs protected mice from a lethal VV challenge by passive immunization. Thus, epitopes that are present on B5 but not on B6 would generate an antibody response that would not recognize B6. Assuming that B6 contains similar variola virus-specific epitopes, our data suggest that a subunit vaccine using the variola virus homologues might exhibit improved protective efficacy against smallpox.
Epstein-Barr Virus Induces MCP-1 Secretion by Human Monocytes via TLR2
Authors: Gaudreault Eric , Fiola Stéphanie , Olivier Martin , Gosselin Jean 
Published: May 2007

ABSTRACTEpstein-Barr virus (EBV) is a gammaherpesvirus infecting the majority of the human adult population in the world. TLR2, a member of the Toll-like receptor (TLR) family, has been implicated in the immune responses to different viruses including members of the herpesvirus family, such as human cytomegalovirus, herpes simplex virus type 1, and varicella-zoster virus. In this report, we demonstrate that infectious and UV-inactivated EBV virions lead to the activation of NF-κB through TLR2 using HEK293 cells cotransfected with TLR2-expressing vector along with NF-κB-Luc reporter plasmid. NF-κB activation in HEK293-TLR2 cells (HEK293 cells transfected with TLR2) by EBV was not enhanced by the presence of CD14. The effect of EBV was abrogated by pretreating HEK293-TLR2 cells with blocking anti-TLR2 antibodies or by preincubating viral particles with neutralizing anti-EBV antibodies 72A1. In addition, EBV infection of primary human monocytes induced the release of MCP-1 (monocyte chemotactic protein 1), and the use of small interfering RNA targeting TLR2 significantly reduced such a chemokine response to EBV. Taken together, these results indicate that TLR2 may be an important pattern recognition receptor in the immune response directed against EBV infection.
Coupled Translation of the Second Open Reading Frame of M2 mRNA Is Sequence Dependent and Differs Significantly within the Subfamily
Pneumovirinae
Authors: Gould Phillip Spencer , Easton Andrew John 
Published: May 2007

ABSTRACT
Coupled translation, first described in the M2 gene of pneumovirus respiratory syncytial virus (RSV), is an alternative mechanism of translational initiation in which the ribosomes which translate the first (M2-1) open reading frame (ORF) move a short distance upstream after termination and reinitiate translation from a second (M2-2) overlapping ORF. Here, we show that the same mechanism occurs in two closely related viruses, avian pneumovirus (APV) and pneumonia virus of mice (PVM), although with markedly different efficiencies. To identify the reasons for the variation in efficiency of coupled expression between RSV and APV, we used chimeric M2-1 genes containing different lengths of the M2-1 ORF from each virus. An essential component allowing coupled expression in the chimeras was a segment from the RSV M2-1 coding region containing a high degree of secondary structure. Additional sequences at the 5′ end of the RSV M2-1 ORF also promoted coupled translation when the region with high levels of secondary structure was present. These data indicate that at least two distant parts of the mRNA transcript, together with a suitable overlapping region, are involved in the coupling process. Replacement of the last 102 nucleotides of the RSV M2-1 ORF with the equivalent APV sequence showed identical levels of coupled translation. Thus, the overlapping region can direct the ribosome back onto the start codon of the second ORF while the upstream coding sequence of the M2-1 ORF determines the levels of coupled expression.
Memory CD4
+
T-Lymphocyte Loss and Dysfunction during Primary Simian Immunodeficiency Virus Infection
Authors: Sun Yue , Permar Sallie R. , Buzby Adam P. , Letvin Norman L. 
Published: May 2007

ABSTRACT

It has long been appreciated that CD4
+
T lymphocytes are dysfunctional in human immunodeficiency virus (HIV)/simian immunodeficiency virus (SIV)-infected individuals, and it has recently been shown that HIV/SIV infections are associated with a dramatic early destruction of memory CD4
+
T lymphocytes. However, the relative contributions of CD4
+
T-lymphocyte dysfunction and loss to immune dysregulation during primary HIV/SIV infection have not been fully elucidated. In the current study, we evaluated CD4
+
T lymphocytes and their functional repertoire during primary SIVmac251 infection in rhesus monkeys. We show that the extent of loss of memory CD4
+
T lymphocytes and staphylococcal enterotoxin B-stimulated cytokine production by total CD4
+
T lymphocytes during primary SIVmac251 infection is tightly linked in a cohort of six rhesus monkeys to set point plasma viral RNA levels, with greater loss and dysfunction being associated with higher steady-state viral replication. Moreover, in exploring the mechanism underlying this phenomenon, we demonstrate that the loss of functional CD4
+
T lymphocytes during primary SIVmac251 infection is associated with both a selective depletion of memory CD4
+
T cells and a loss of the functional capacity of the memory CD4
+
T lymphocytes that escape viral destruction.
Expression of Extremely Low Levels of Thymidine Kinase from an Acyclovir-Resistant Herpes Simplex Virus Mutant Supports Reactivation from Latently Infected Mouse Trigeminal Ganglia
Authors: Besecker Michael I. , Furness Caroline L. , Coen Donald M. , Griffiths Anthony 
Published: May 2007

ABSTRACT
A single-cytosine-deletion in the herpes simplex virus gene encoding thymidine kinase (TK) was previously found in an acyclovir-resistant clinical isolate. A laboratory strain engineered to carry this mutation did not generate sufficient TK activity for detection by plaque autoradiography, which detected 0.25% wild-type activity. However, a drug sensitivity assay suggested that extremely low levels of TK are generated by this virus. The virus was estimated to express 0.09% of wild-type TK activity via a ribosomal frameshift 24 nucleotides upstream of the mutation. Remarkably, this appeared to be sufficient active TK to support a low level of reactivation from latently infected mouse trigeminal ganglia.
Pretreatment Sequence Diversity Differences in the Full-Length Hepatitis C Virus Open Reading Frame Correlate with Early Response to Therapy
Authors: Donlin Maureen J. , Cannon Nathan A. , Yao Ermei , Li Jia , Wahed Abdus , Taylor Milton W. , Belle Steven H. , Di Bisceglie Adrian M. , Aurora Rajeev , Tavis John E. 
Published: May 2007

ABSTRACT
Pegylated alpha interferon and ribavirin therapy for hepatitis C virus (HCV) genotype 1 infection fails for half of Caucasian American patients (CA) and more often for African Americans (AA). The reasons for these low response rates are unknown. HCV is highly genetically variable, but it is unknown how this variability affects response to therapy. To assess effects of viral diversity on response to therapy, the complete pretreatment genotype 1 HCV open reading frame was sequenced using samples from 94 participants in the Virahep-C study. Sequences from patients with >3.5 log declines in viral RNA levels by day 28 (marked responders) were more variable than those from patients with declines of <1.4 log (poor responders) in NS3 and NS5A for genotype 1a and in core and NS3 for genotype 1b. These correlations remained when all T-cell epitopes were excluded, indicating that these differences were not due to differential immune selection. When the sequences were compared by race of the patients, higher diversity in CA patients was found in E2 and NS2 but only for genotype 1b. Core, NS3, and NS5A can block the action of alpha interferon in vitro; hence, these genetic patterns are consistent with multiple amino acid variations independently impairing the function of HCV proteins that counteract interferon responses in humans, resulting in HCV strains with variable sensitivity to therapy. No evidence was found for novel HCV strains in the AA population, implying that AA patients may be infected with a higher proportion of the same resistant strains that are found in CA patients.
A Herpes Simplex Virus gD-YFP Fusion Glycoprotein Is Transported Separately from Viral Capsids in Neuronal Axons
Authors: Snyder Aleksandra , Bruun Birgitte , Browne Helena M. , Johnson David C. 
Published: May 2007

ABSTRACT
Two models describing how alphaherpesviruses exit neurons differ with respect to whether nucleocapsids and envelope glycoproteins travel toward axon termini separately or as assembled enveloped virions. Recently, a pseudorabies virus glycoprotein D (gD)-green fluorescent protein fusion was found to colocalize with viral capsids, supporting anterograde transport of enveloped virions. Previous antibody staining experiments demonstrated that herpes simplex virus (HSV) glycoproteins and capsids are separately transported in axons. Here, we generated an HSV expressing a gD-yellow fluorescent protein (YFP) fusion and found that gD-YFP and capsids were transported separately in neuronal axons. Anti-gD antibodies colocalized with gD-YFP, indicating that gD-YFP behaves like wild-type HSV gD.
The role of physical examinations in studies of musculoskeletal disorders of the elbow
Authors: Kryger Ann Isabel , Lassen Christina Funch , Andersen Johan Hviid 
Published: May 2007

Objectives:To present data on pain and physical findings from the elbow region, and to discuss the role of diagnostic criteria in epidemiological studies of epicondylitis.Methods:From a cohort of computer workers a subgroup of 1369 participants, who reported at least moderate pain in the neck and upper extremities, were invited to a standardised physical examination. Two independent physical examinations were performedone blinded and one not blinded to the medical history. Information concerning musculoskeletal symptoms was obtained by a baseline questionnaire and a similar questionnaire completed on the day of examination.Results:349 participants met the authors criteria for being an arm case and 249 were elbow cases. Among the 1369 participants the prevalence of at least mild palpation tenderness and indirect tenderness at the lateral epicondyle was 5.8. The occurrence of physical findings increased markedly by level of pain score. Only about one half with physical findings fulfilled the authors pain criteria for having lateral epicondylitis. A large part with physical findings reported no pain at all in the elbow in any of the two questionnaires, 28 and 22, respectively. Inter-examiner reliability between blinded and not blinded examination was found to be low kappa value 0.340.40.Conclusion:Very few with at least moderate pain in the elbow region met common specific criteria for lateral epicondylitis. The occurrence of physical findings increased markedly by level of pain score and the associations were strongest with pain intensity scores given just before the examination. Physical signs were commonly found in subjects with no pain complaints. No further impact was achieved if the physical examination was not blinded to the medical history. Furthermore, the authors propose that pain, clinical signs and disability are studied as separate outcomes, and that the diagnoses of lateral epicondylitis should be used only for cases with classical signs of inflammation reflected by severe pain, which for example conveys some disability.
Field protection effectiveness of chemical protective suits and gloves evaluated by biomonitoring
Authors: Chang F K , Chen M L , Cheng S F , Shih T S , Mao I F 
Published: May 2007

Objectives:To determine the effectiveness of protective suits and gloves by biomonitoring.Methods:Fifteen male spray painters at a ship coating factory were studied for two weeks. Workers wore no protective clothing during the first week and wore protective suits and gloves during the second week. Sampling was conducted on four consecutive working days each week. Ethyl benzene and xylene in the air were collected by using 3M 3500 organic vapour monitors. Urine was collected before and after each work shift.Results:Urinary mandelic acid MA and methyl hippuric acid MHA levels were divided by the personal exposure concentrations of ethyl benzene and xylene, respectively. Mean SE corrected MA and MHA concentrations in the first week were 1.07 0.18 and 2.66 0.68 mg/g creatinine/mg/m3, and concentrations in the second week were 0.50 0.12 and 1.76 0.35 mg/g creatinine/mg/m3in the second week, respectively. Both MA and MHA concentrations in the second week when spray painters wore protective suits and gloves were lower than in the first week, respectively p<0.001, p0.011. Mean decrease in MA and MHA biomarkers were 69 and 49, respectively.Conclusion:This study successfully evaluated the effectiveness of chemical protective suits and gloves by using biomarkers as urinary MA and MHA. This method is feasible for determining the performance of workers wearing personal protective equipment. Moreover, the experimental results suggest that dermal exposure may be the major contributor to total body burden of solvents in spray painters without protective suits and gloves.
Mineral dust exposure in young Indian adults: an effect on lung growth?
Authors: Green D A , McAlpine G , Semple S , Cowie H , Seaton A 
Published: May 2007

Background/objective:Long-term occupational exposure to mineral dusts including quartz is known to cause pneumoconiosis but little is known about the effects of such exposure during the period of lung development in childhood. The objective of this study was to investigate the effects on young people of exposure to mineral dusts.Methods:We carried out a cross-sectional study of a group of 81 Indian stone crushers (average age 21) and a control group of 72 nearby slum dwellers (average age 21) in order to assess dust exposures, respiratory symptoms and lung function.Results:Stone crushers’ corrected 8-hour time weighted average concentrations of total inhalable dust, respirable dust and respirable crystalline silica were 143 mg/m3, 39.7 mg/m3and 2.29 mg/m3respectively, which are 14, 10 and 23 times the UK workplace exposure limits, respectively. Both groups also had high exposures to outdoor and general domestic particulate matter. Compared to controls, stone crushers had significantly more symptoms, lower levels of forced vital capacity (FVC) and forced expiratory time (FET) and higher levels of forced expiratory volume in one second (FEV1)/FVC ratio and mid-expiratory flow rate (MEF). There were no differences in FEV1levels. Symptoms and lung function were not associated with time lived on site or time spent in job. A selection of chest radiographs showed no silicosis by International Labour Office (ILO) criteria.Conclusion:Individuals exposed to mixed mineral dust in childhood and early adult life had excess symptoms and reduced vital capacity without airflow obstruction compared with control subjects. We interpret this as evidence of stunting of lung growth caused by exposure to respirable dust.
Loss of Betaglycan Expression in Ovarian Cancer: Role in Motility and Invasion
Authors: Hempel Nadine , How Tam , Dong Mei , Murphy Susan K. , Fields Timothy A. , Blobe Gerard C. 
Published: May 2007

Abstract
The transforming growth factor-β (TGF-β) superfamily members, TGF-β, activin, and inhibin, all have prominent roles in regulating normal ovarian function. Betaglycan, or the type III TGF-β receptor, is a coreceptor that regulates TGF-β, activin, and inhibin signaling. Here, we show that betaglycan expression is frequently decreased or lost in epithelial derived ovarian cancer at both the mRNA and protein level, with the degree of loss correlating with tumor grade. Treatment of ovarian cancer cell lines with the methyltransferase inhibitor 5-aza-2-deoxycytidine and the histone deacetylase inhibitor trichostatin A resulted in significant synergistic induction of betaglycan message levels and increased betaglycan protein expression, indicating that epigenetic silencing may play a role in the loss of betaglycan expression observed in ovarian cancer. Although restoring betaglycan expression in Ovca429 ovarian cancer cells is not sufficient to restore TGF-β–mediated inhibition of proliferation, betaglycan significantly inhibits ovarian cancer cell motility and invasiveness. Furthermore, betaglycan specifically enhances the antimigratory effects of inhibin and the ability of inhibin to repress matrix metalloproteinase levels in these cells. These results show, for the first time, epigenetic regulation of betaglycan expression in ovarian cancer, and a novel role for betaglycan in regulating ovarian cancer motility and invasiveness. [Cancer Res 2007;67(11):5231–8]