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adjuvant-like effect of vaccinia virus 14k protein: a case study with malaria vaccine based on the circumsporozoite protein.development of subunit vaccines for malaria that elicit a strong, long-term memory response is an intensive area of research, with the focus on improving the immunogenicity of a circumsporozoite (cs) protein-based vaccine. in this study, we found that a chimeric protein, formed by fusing vaccinia virus protein 14k (a27) to the cs of plasmodium yoelii, induces strong effector memory cd8(+) t cell responses in addition to high-affinity abs when used as a priming agent in the absence of any adjuvan ...201222615208
antiviral activities and putative identification of compounds in microbial extracts from the hawaiian coastal waters.marine environments are a rich source of significant bioactive compounds. the hawaiian archipelago, located in the middle of the pacific ocean, hosts diverse microorganisms, including many endemic species. thirty-eight microbial extracts from hawaiian coastal waters were evaluated for their antiviral activity against four mammalian viruses including herpes simplex virus type one (hsv-1), vesicular stomatitis virus (vsv), vaccinia virus and poliovirus type one (poliovirus-1) using in vitro cell c ...201222611351
genome-wide analysis of polymorphisms associated with cytokine responses in smallpox vaccine recipients.the role that genetics play in response to infection or disease is becoming increasingly clear as we learn more about immunogenetics and host-pathogen interactions. here we report a genome-wide analysis of the effects of host genetic variation on cytokine responses to vaccinia virus stimulation in smallpox vaccine recipients. our data show that vaccinia stimulation of immune individuals results in secretion of inflammatory and th1 cytokines. we identified multiple snps significantly associated w ...201222610502
innate immune response of human plasmacytoid dendritic cells to poxvirus infection is subverted by vaccinia e3 via its z-dna/rna binding domain.plasmacytoid dendritic cells (pdcs) play important roles in antiviral innate immunity by producing type i interferon (ifn). in this study, we assess the immune responses of primary human pdcs to two poxviruses, vaccinia and myxoma virus. vaccinia, an orthopoxvirus, was used for immunization against smallpox, a contagious human disease with high mortality. myxoma virus, a leporipoxvirus, causes lethal disease in rabbits, but is non-pathogenic in humans. we report that myxoma virus infection of hu ...201222606294
the death receptor 3/tl1a pathway is essential for efficient development of antiviral cd4⁺ and cd8⁺ t-cell immunity.death receptor 3 (dr3, tnfrsf25), the closest family relative to tumor necrosis factor receptor 1, promotes cd4(+) t-cell-driven inflammatory disease. we investigated the in vivo role of dr3 and its ligand tl1a in viral infection, by challenging dr3-deficient (dr3(ko)) mice and their dr3(wt) littermates with the β-herpesvirus murine cytomegalovirus or the poxvirus vaccinia virus. the phenotype and function of splenic t-cells were analyzed using flow cytometry and molecular biological techniques. ...201222593543
analysis of variola and vaccinia virus neutralization assays for smallpox vaccines.possible smallpox reemergence drives research for third-generation vaccines that effectively neutralize variola virus. a comparison of neutralization assays using different substrates, variola and vaccinia (dryvax and modified vaccinia ankara [mva]), showed significantly different 90% neutralization titers; dryvax underestimated while mva overestimated variola neutralization. third-generation vaccines may rely upon neutralization as a correlate of protection.201222593237
pd-1/pd-l1 blockade can enhance hiv-1 gag-specific t cell immunity elicited by dendritic cell-directed lentiviral vaccines.exhaustion of cd8(+) t cells and upregulation of programmed death 1 (pd-1), a negative regulator of t cell activation, are characteristic features of individuals chronically infected with human immunodeficiency virus type 1. in a previous study, we showed in mice that a dendritic cell-directed lentiviral vector (dclv) system encoding the human immunodeficiency virus (hiv)-1 gag protein was an efficient vaccine modality to induce a durable gag-specific t cell immune response. in this study, we de ...201222588271
synthesis and antiviral evaluation of c5-substituted-(1,3-diyne)-2'-deoxyuridines.starting from acetylated 5-ethynyl-2'-deoxyuridine (3), 14 hitherto unknown c5-substituted-(1,3-diyne)-2'-deoxyuridines (with cyclopropyl, hydroxymethyl, methylcyclopentane, p-(substituted)phenyl and disubstituted-phenyl substituents) have been synthesized via a nickel-copper catalyzed c-h activation between two terminal alkynes, in yields ranging from 19% to 67%. their antiviral activities were measured against a large number of dna and rna viruses including herpes simplex virus type 1 and type ...201222578783
position on donors and smallpox vaccine: a committee opinion.although there is presently no definitive evidence linking vaccinia virus transmission through reproductive cells, sart/asrm accordingly recommends that art practitioners consider deferring donors who have recently received smallpox vaccine or contracted symptomatic vaccinia virus infection through close contact with a vaccine recipient (until after the vaccine or infectious scab has spontaneously separated). good donor practice further suggests that donors who are not in good health, including ...201222537381
systems analysis of mva-c induced immune response reveals its significance as a vaccine candidate against hiv/aids of clade c.based on the partial efficacy of the hiv/aids thai trial (rv144) with a canarypox vector prime and protein boost, attenuated poxvirus recombinants expressing hiv-1 antigens are increasingly sought as vaccine candidates against hiv/aids. here we describe using systems analysis the biological and immunological characteristics of the attenuated vaccinia virus ankara strain expressing the hiv-1 antigens env/gag-pol-nef of hiv-1 of clade c (referred as mva-c). mva-c infection of human monocyte derive ...201222536391
entry of human papillomavirus type 16 by actin-dependent, clathrin- and lipid raft-independent endocytosis.infectious endocytosis of incoming human papillomavirus type 16 (hpv-16), the main etiological agent of cervical cancer, is poorly characterized in terms of cellular requirements and pathways. conflicting reports attribute hpv-16 entry to clathrin-dependent and -independent mechanisms. to comprehensively describe the cell biological features of hpv-16 entry into human epithelial cells, we compared hpv-16 pseudovirion (psv) infection in the context of cell perturbations (drug inhibition, sirna si ...201222536154
loss of cytoskeletal transport during egress critically attenuates ectromelia virus infection in vivo.egress of wrapped virus (wv) to the cell periphery following vaccinia virus (vacv) replication is dependent on interactions with the microtubule motor complex kinesin-1 and is mediated by the viral envelope protein a36. here we report that ectromelia virus (ectv), a related orthopoxvirus and the causative agent of mousepox, encodes an a36 homologue (ectv-mos-142) that is highly conserved despite a large truncation at the c terminus. deleting the ectv a36r gene leads to a reduction in the number ...201222532690
mutations conferring resistance to viral dna polymerase inhibitors in camelpox virus give different drug-susceptibility profiles in vaccinia virus.cidofovir or (s)-hpmpc is one of the three antiviral drugs that might be used for the treatment of orthopoxvirus infections. (s)-hpmpc and its 2,6-diaminopurine counterpart, (s)-hpmpdap, have been described to select, in vitro, for drug resistance mutations in the viral dna polymerase (e9l) gene of vaccinia virus (vacv). here, to extend our knowledge of drug resistance development among orthopoxviruses, we selected, in vitro, camelpox viruses (cmlv) resistant to (s)-hpmpdap and identified a sing ...201222532673
comparable polyfunctionality of ectromelia virus- and vaccinia virus-specific murine t cells despite markedly different in vivo replication and pathogenicity.vaccinia virus (vacv) stimulates long-term immunity against highly pathogenic orthopoxvirus infection of humans (smallpox) and mice (mousepox [ectromelia virus {ectv}]) despite the lack of a natural host-pathogen relationship with either of these species. previous research revealed that vacv is able to induce polyfunctional cd8(+) t-cell responses after immunization of humans. however, the degree to which the functional profile of t cells induced by vacv is similar to that generated during natur ...201222532670
vaccine efficacy against malaria by the combination of porcine parvovirus-like particles and vaccinia virus vectors expressing cs of plasmodium.with the aim to develop an efficient and cost-effective approach to control malaria, we have generated porcine parvovirus-like particles (ppv-vlps) carrying the cd8(+) t cell epitope (syvpsaeqi) of the circumsporozoite (cs) protein from plasmodium yoelii fused to the ppv vp2 capsid protein (ppv-pycs), and tested in prime/boost protocols with poxvirus vectors for efficacy in a rodent malaria model. as a proof-of concept, we have characterized the anti-cs cd8(+) t cell response elicited by these h ...201222529915
modulation of the myxoma virus plaque phenotype by vaccinia virus protein f11.vaccinia virus (vacv) produces large plaques consisting of a rapidly expanding ring of infected cells surrounding a lytic core, whereas myxoma virus (myxv) produces small plaques that resemble a focus of transformed cells. this is odd, because bioinformatics suggests that myxv carries homologs of nearly all of the genes regulating orthopoxvirus attachment, entry, and exit. so why does myxv produce foci? one notable difference is that myxv-infected cells produce few of the actin microfilaments th ...201222514354
theranostic potential of oncolytic vaccinia virus.biological cancer therapies, such as oncolytic, or replication-selective viruses have advantages over traditional therapeutics as they can employ multiple different mechanisms to target and destroy cancers (including direct cell lysis, immune activation and vascular collapse). this has led to their rapid recent clinical development. however this also makes their pre-clinical and clinical study complex, as many parameters may affect their therapeutic potential and so defining reason for treatment ...201222509200
mhc class i antigen processing distinguishes endogenous antigens based on their translation from cellular vs. viral mrna.to better understand the generation of mhc class i-associated peptides, we used a model antigenic protein whose proteasome-mediated degradation is rapidly and reversibly controlled by shield-1, a cell-permeant drug. when expressed from a stably transfected gene, the efficiency of antigen presentation is ~2%, that is, one cell-surface mhc class i-peptide complex is generated for every 50 folded source proteins degraded upon shield-1 withdrawal. by contrast, when the same protein is expressed by v ...201222509014
the dc receptor dngr-1 mediates cross-priming of ctls during vaccinia virus infection in mice.in order to prime t cells, dcs integrate signals emanating directly from pathogens and from their noxious action on the host. dngr-1 (clec9a) is a dc-restricted receptor that detects dead cells. therefore, we investigated the possibility that dngr-1 affects immunity to cytopathic viruses. dngr-1 was essential for cross-presentation of dying vaccinia virus-infected (vacv-infected) cells to cd8(+) t cells in vitro. following injection of vacv or vacv-infected cells into mice, dngr-1 detected the l ...201222505455
integrin β1 mediates vaccinia virus entry through activation of pi3k/akt signaling.vaccinia virus has a broad range of infectivity in many cell lines and animals. although it is known that the vaccinia mature virus binds to cell surface glycosaminoglycans and extracellular matrix proteins, whether additional cellular receptors are required for virus entry remains unclear. our previous studies showed that the vaccinia mature virus enters through lipid rafts, suggesting the involvement of raft-associated cellular proteins. here we demonstrate that one lipid raft-associated prote ...201222496232
poxvirus infection-associated downregulation of c-type lectin-related-b prevents nk cell inhibition by nk receptor protein-1b.innate immune recognition of virus-infected cells includes nk cell detection of changes to endogenous cell-surface proteins through inhibitory receptors. one such receptor system is the nk cell receptor protein-1b (nkr-p1b) and its ligand c-type lectin-related-b (clr-b). nkr-p1b and clr-b are encoded within the nk cell gene complex, a locus that has been linked to strain-dependent differences in susceptibility to infection by poxviruses. in this study, we report the impact of vaccinia virus (vv) ...201222491247
combined costimulatory and leukocyte functional antigen-1 blockade prevents transplant rejection mediated by heterologous immune memory alloresponses.recent evidence suggests that alloreactive memory t cells are generated by the process of heterologous immunity, whereby memory t cells arising in response to pathogen infection crossreact with donor antigens. because of their diminished requirements for costimulation during recall, these pathogen-elicited allocrossreactive memory t cells are of particular clinical importance, especially given the emergence of costimulatory blockade as a transplant immunosuppression strategy.201222475765
in vitro transcription and capping of gaussia luciferase mrna followed by hela cell transfection.in vitro transcription is the synthesis of rna transcripts by rna polymerase from a linear dna template containing the corresponding promoter sequence (t7, t3, sp6) and the gene to be transcribed (figure 1a). a typical transcription reaction consists of the template dna, rna polymerase, ribonucleotide triphosphates, rnase inhibitor and buffer containing mg(2+) ions. large amounts of high quality rna are often required for a variety of applications. use of in vitro transcription has been reported ...201222473375
stability of vaccinia-vectored recombinant oral rabies vaccine under field conditions: a 3-year study.rabies is an incurable zoonotic disease caused by rabies virus, a member of the rhabdovirus family. it is transmitted through the bite of an infected animal. control methods, including oral rabies vaccination (orv) programs, have led to a reduction in the spread and prevalence of the disease in wildlife. this study evaluated the stability of raboral, a recombinant vaccinia virus vaccine that is used in oral rabies vaccination programs. the vaccine was studied in various field microenvironments i ...201122468025
poxvirus host cell entry.poxviruses are characterized by their large size, complex composition, and cytoplasmic life cycle. they produce two types of infectious particles: mature virions (mvs) and extracellular virions (evs). both mvs and evs of vaccinia virus, the model poxvirus, take advantage of host cell endocytosis for internalization: they activate macropinocytosis-the most suitable form of endocytosis for large particles. although largely dependent on the same cellular machinery, mv and ev entry differs with rega ...201122440962
a novel high-throughput vaccinia virus neutralization assay and preexisting immunity in populations from different geographic regions in china.pre-existing immunity to vaccinia tian tan virus (vtt) resulting from a large vaccination campaign against smallpox prior to the early 1980s in china, has been a major issue for application of vtt-vector based vaccines. it is essential to establish a sensitive and high-throughput neutralization assay to understand the epidemiology of vaccinia-specific immunity in current populations in china.201222438922
increased protection from vaccinia virus infection in mice genetically prone to lymphoproliferative disorders.mutations in the genes that encode fas or fas ligand (fasl) can result in poor restraints on lymphocyte activation and in increased susceptibility to autoimmune disorders. because these mutations portend a continuously activated immune state, we hypothesized that they might in some cases confer resistance to infection. to examine this possibility, the immune response to, morbidity caused by, and clearance of vaccinia virus (vacv) western reserve was examined in 5- to 7-week-old fas mutant (lpr) ...201222438562
molecular genetic and biochemical characterization of the vaccinia virus i3 protein, the replicative single-stranded dna binding protein.vaccinia virus, the prototypic poxvirus, efficiently and faithfully replicates its ∼200-kb dna genome within the cytoplasm of infected cells. this intracellular localization dictates that vaccinia virus encodes most, if not all, of its own dna replication machinery. included in the repertoire of viral replication proteins is the i3 protein, which binds to single-stranded dna (ssdna) with great specificity and stability and has been presumed to be the replicative ssdna binding protein (ssb). we s ...201222438556
formation of orthopoxvirus cytoplasmic a-type inclusion bodies and embedding of virions are dynamic processes requiring microtubules.in cells infected with some orthopoxviruses, numerous mature virions (mvs) become embedded within large, cytoplasmic a-type inclusions (atis) that can protect infectivity after cell lysis. atis are composed of an abundant viral protein called atip, which is truncated in orthopoxviruses such as vaccinia virus (vacv) that do not form atis. to study ati formation and occlusion of mvs within atis, we used recombinant vacvs that express the cowpox full-length atip or we transfected plasmids encoding ...201222438543
use of interferon-γ enzyme-linked immunospot assay to characterize novel t-cell epitopes of human papillomavirus.a protocol has been developed to overcome the difficulties of isolating and characterizing rare t cells specific for pathogens, such as human papillomavirus (hpv), that cause localized infections. the steps involved are identifying region(s) of hpv proteins that contain t-cell epitope(s) from a subject, selecting for the peptide-specific t cells based on interferon-γ (ifn-γ) secretion, and growing and characterizing the t-cell clones (fig. 1). subject 1 was a patient who was recently diagnosed w ...201222434036
targeting pediatric cancer stem cells with oncolytic virotherapy.cancer stem cells (cscs), also termed "cancer-initiating cells" or "cancer progenitor cells," which have the ability to self-renew, proliferate, and maintain the neoplastic clone, have recently been discovered in a wide variety of pediatric tumors. these cscs are thought to be responsible for tumorigenesis and tumor maintenance, aggressiveness, and recurrence due to inherent resistance to current treatment modalities such as chemotherapy and radiation. oncolytic virotherapy offers a novel, targe ...201222430386
in vitro characterization of a nineteenth-century therapy for smallpox.in the nineteenth century, smallpox ravaged through the united states and canada. at this time, a botanical preparation, derived from the carnivorous plant sarracenia purpurea, was proclaimed as being a successful therapy for smallpox infections. the work described characterizes the antipoxvirus activity associated with this botanical extract against vaccinia virus, monkeypox virus and variola virus, the causative agent of smallpox. our work demonstrates the in vitro characterization of sarracen ...201222427855
uv-inactivated vaccinia virus (vv) in a multi-envelope dna-vv-protein (dvp) hiv-1 vaccine protects macaques from lethal challenge with heterologous shiv.the pandemic of hiv-1 has continued for decades, yet there remains no licensed vaccine. previous research has demonstrated the effectiveness of a multi-envelope, multi-vectored hiv-1 vaccine in a macaque-shiv model, illustrating a potential means of combating hiv-1. specifically, recombinant dna, vaccinia virus (vv) and purified protein (dvp) delivery systems were used to vaccinate animals with dozens of antigenically distinct hiv-1 envelopes for induction of immune breadth. the vaccinated anima ...201222425790
cd27 stimulation promotes the frequency of il-7 receptor-expressing memory precursors and prevents il-12-mediated loss of cd8(+) t cell memory in the absence of cd4(+) t cell help.fully functional cd8(+) t cell memory is highly dependent upon cd4(+) t cell support. cd4(+) t cells play a critical role in inducing the expression of cd70, the ligand for cd27, on dendritic cells. in this study, we demonstrate that cd27 stimulation during primary cd8(+) t cell responses regulates the ability to mount secondary cd8(+) t cell responses. cd27 stimulation during vaccinia and dendritic cell immunization controls the expression of the il-7r (cd127), which has been shown to be necess ...201222422886
the amino terminus of the vaccinia virus e3 protein is necessary to inhibit the interferon response.vaccinia virus (vacv) encodes a multifunctional protein, e3l, that is necessary for interferon (ifn) resistance in cells in culture. interferon resistance has been mapped to the well-characterized carboxy terminus of e3l, which contains a conserved double-stranded rna binding domain. the amino terminus of e3l has a z-form nucleic acid binding domain, which has been shown to be dispensable for replication and ifn resistance in hela and rk13 cells; however, a virus expressing e3l deleted of the am ...201222419806
removal of vaccinia virus genes that block interferon type i and ii pathways improves adaptive and memory responses of the hiv/aids vaccine candidate nyvac-c in mice.poxviruses encode multiple inhibitors of the interferon (ifn) system, acting at different levels and blocking the induction of host defense mechanisms. two viral gene products, b19 and b8, have been shown to act as decoy receptors of type i and type ii ifns, blocking the binding of ifn to its receptor. since ifn plays a major role in innate immune responses, in this investigation we asked to what extent the viral inhibitors of the ifn system impact the capacity of poxvirus vectors to activate im ...201222419805
six-transmembrane topology for golgi anti-apoptotic protein (gaap) and bax inhibitor 1 (bi-1) provides model for the transmembrane bax inhibitor-containing motif (tmbim) family.the golgi anti-apoptotic protein (gaap) is a hydrophobic golgi protein that regulates intracellular calcium fluxes and apoptosis. gaap is highly conserved throughout eukaryotes and some strains of vaccinia virus (vacv) and camelpox virus. based on sequence, phylogeny, and hydrophobicity, gaaps were classified within the transmembrane bax inhibitor-containing motif (tmbim) family. tmbim members are anti-apoptotic and were predicted to have seven-transmembrane domains (tmds). however, topology pre ...201222418439
synthesis and antiviral activity of 6-deoxycyclopropavir, a new prodrug of cyclopropavir.synthesis of 6-deoxycyclopropavir (10), a prodrug of cyclopropavir (1) and its in vitro and in vivo antiviral activity is described. 2-amino-6-chloropurine methylenecyclopropane 13 was transformed to its 6-iodo derivative 14 which was reduced to prodrug 10. it is converted to cyclopropavir (1) by the action of xanthine oxidase and this reaction can also occur in vivo. compound 10 lacked significant in vitro activity against human cytomegalovirus (hcmv), human herpes virus 1 and 2 (hsv-1 and hsv- ...201222417649
reverse genetics of sars-related coronavirus using vaccinia virus-based recombination.severe acute respiratory syndrome (sars) is a zoonotic disease caused by sars-related coronavirus (sars-cov) that emerged in 2002 to become a global health concern. although the original outbreak was controlled by classical public health measures, there is a real risk that another sars-cov could re-emerge from its natural reservoir, either in its original form or as a more virulent or pathogenic strain; in which case, the virus would be difficult to control in the absence of any effective antivi ...201222412934
orthopoxvirus targets for the development of new antiviral agents.investments in the development of new drugs for orthopoxvirus infections have fostered new avenues of research, provided an improved understanding of orthopoxvirus biology and yielded new therapies that are currently progressing through clinical trials. these broad-based efforts have also resulted in the identification of new inhibitors of orthopoxvirus replication that target many different stages of viral replication cycle. this review will discuss progress in the development of new anti-poxvi ...201222406470
delivery of echinococcus granulosus antigen eg95 to mice and sheep using recombinant vaccinia virus.the tapeworm echinococcus granulosus is the causative agent of hydatid disease and affects sheep, cattle, dogs and humans worldwide. it has a two-stage life cycle existing as worms in the gut of infected dogs (definitive host) and as cysts in herbivores and humans (intermediate host). the disease is debilitating and can be life threatening where the cysts interfere with organ function. interruption of the hydatid life cycle in the intermediate host by vaccination may be a way to control the dise ...201222404504
the myristate moiety and amino terminus of vaccinia virus l1 constitute a bipartite functional region needed for entry.vaccinia virus (vacv) l1 is a myristoylated envelope protein which is required for cell entry and the fusion of infected cells. l1 associates with members of the entry-fusion complex (efc), but its specific role in entry has not been delineated. we recently demonstrated (foo ch, et al., virology 385:368-382, 2009) that soluble l1 binds to cells and blocks entry, suggesting that l1 serves as the receptor-binding protein for entry. our goal is to identify the structural domains of l1 which are ess ...201222398293
vaccinia virus a6 is essential for virion membrane biogenesis and localization of virion membrane proteins to sites of virion assembly.poxvirus acquires its primary envelope through a process that is distinct from those of other enveloped viruses. the molecular mechanism of this process is poorly understood, but several poxvirus proteins essential for the process have been identified in studies of vaccinia virus (vacv), the prototypical poxvirus. previously, we identified vacv a6 as an essential factor for virion morphogenesis by studying a temperature-sensitive mutant with a lesion in a6. here, we further studied a6 by constru ...201222398288
skin infection generates non-migratory memory cd8+ t(rm) cells providing global skin immunity.protective t-cell memory has long been thought to reside in blood and lymph nodes, but recently the concept of immune memory in peripheral tissues mediated by resident memory t (t(rm)) cells has been proposed. here we show in mice that localized vaccinia virus (vacv) skin infection generates long-lived non-recirculating cd8(+) skin t(rm) cells that reside within the entire skin. these skin t(rm) cells are potent effector cells, and are superior to circulating central memory t (t(cm)) cells at pr ...201222388819
improving the mva vaccine potential by deleting the viral gene coding for the il-18 binding protein.modified vaccinia ankara (mva) is an attenuated strain of vaccinia virus (vacv) currently employed in many clinical trials against hiv/aids and other diseases. mva still retains genes involved in host immune response evasion, enabling its optimization by removing some of them. the aim of this study was to evaluate cellular immune responses (cir) induced by an il-18 binding protein gene (c12l) deleted vector (mvaδc12l).201222384183
preferential replication of systemically delivered oncolytic vaccinia virus in focally irradiated glioma xenografts.radiotherapy is part of the standard of care in high-grade gliomas but its outcomes remain poor. integrating oncolytic viruses with standard anticancer therapies is an area of active investigation. the aim of this study was to determine how tumor-targeted ionizing radiation (ir) could be combined with systemically delivered oncolytic vaccinia virus.201222379115
validation of an immunoperoxidase monolayer assay for total anti-vaccinia virus antibody titration.vaccinia virus (vacv) has been associated with zoonotic exanthemic outbreaks affecting bovids and human beings, with significant public health and economic impacts. rapid and reliable diagnostic methods are needed to detect and epidemiologically monitor antibodies to vacv. the current study describes the development of an immunoperoxidase monolayer assay (ipma) for detection of total vacv antibodies in bovine serum. the assay was validated by comparison with a plaque reduction neutralization tes ...201222379052
effect of serum type and concentration on the expression ofβ-galactosidase in recombinant vaccinia virus infected hela s3 cells.the effect of serum type and concentration on recombinant protein expression in vaccinia virus infected hela s3 cells was studied in both static and suspension culture. a model heterologous protein,β-galactosidase (β-gal), was used. calf and horse sera in the range of 0.5-10%(v/v) were investigated. in static culture, the calf serum concentration did not show any significant influence on the β-gal production which was almost completed within 24h postinfection (pi). higher horse serum concentrati ...199522359015
biological characterization and next-generation genome sequencing of the unclassified cotia virus span232 (poxviridae).cotia virus (cotv) span232 was isolated in 1961 from sentinel mice at cotia field station, são paulo, brazil. attempts to classify cotv within a recognized genus of the poxviridae have generated contradictory findings. studies by different researchers suggested some similarity to myxoma virus and swinepox virus, whereas another investigation characterized cotv span232 as a vaccinia virus strain. because of the lack of consensus, we have conducted an independent biological and molecular character ...201222345477
the lipid raft-associated protein cd98 is required for vaccinia virus endocytosis.mature vaccinia virus (vaccinia mv) infects a broad range of animals in vivo and cell cultures in vitro; however, the cellular receptors that determine vaccinia mv tropism and entry pathways are poorly characterized. here, we performed quantitative proteomic analyses of lipid raft-associated proteins upon vaccinia mv entry into hela cells. we found that a type ii membrane glycoprotein, cd98, is enriched in lipid rafts upon vaccinia mv infection compared to mock-infected hela cells. the knockdown ...201222345471
c7l family of poxvirus host range genes inhibits antiviral activities induced by type i interferons and interferon regulatory factor 1.vaccinia virus (vacv) k1l and c7l function equivalently in many mammalian cells to support vacv replication and antagonize antiviral activities induced by type i interferons (ifns). while k1l is limited to orthopoxviruses, genes that are homologous to c7l are found in diverse mammalian poxviruses. in this study, we showed that the c7l homologues from sheeppox virus and swinepox virus could rescue the replication defect of a vacv mutant deleted of both k1l and c7l (vk1l(-)c7l(-)). interestingly, ...201222345458
effective oncolytic vaccinia therapy for human sarcomas.approximately one fourth of bone and soft-tissue sarcomas recur after prior treatment. glv-1h68 is a recombinant, replication-competent vaccinia virus that has been shown to have oncolytic effects against many human cancer types. we sought to determine whether glv-1h68 could selectively target and lyse a panel of human bone and soft-tissue sarcoma cell lines in vitro and in vivo.201122341347
electrochemical differentiation of epitope-specific aptamers.dna aptamers are promising immunoshielding agents that could protect oncolytic viruses (ovs) from neutralizing antibodies (nabs) and increase the efficiency of cancer treatment. in the present article, we introduce a novel technology for electrochemical differentiation of epitope-specific aptamers (edea) without selecting aptamers against individual antigenic determinants. for this purpose, we selected dna aptamers that can bind noncovalently to an intact oncolytic virus, vaccinia virus (vacv), ...201222324738
a report of 2 cases of myopericarditis after vaccinia virus (smallpox) immunization.to counter the possibility of smallpox being used as a biological weapon, in 2002 the us government restarted a smallpox vaccination campaign. myopericarditis is a possible cardiac complication of smallpox vaccination. we report 2 cases of vaccine-associated myopericarditis in military recruits who were treated at our facility. chest pain, shortness of breath, and electrocardiographic changes of pericarditis, with a recent history of smallpox vaccination, were useful in making the diagnosis of p ...201122324207
neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention.human respiratory syncytial virus (hrsv) is the most important viral agent of pediatric respiratory infections worldwide. the only specific treatment available today is a humanized monoclonal antibody (palivizumab) directed against the f glycoprotein, administered prophylactically to children at very high risk of severe hrsv infections. palivizumab, as most anti-f antibodies so far described, recognizes an epitope that is shared by the two conformations in which hrsv_f can fold, the metastable p ...201222323598
disruption of tnf-α/tnfr1 function in resident skin cells impairs host immune response against cutaneous vaccinia virus infection.one strategy adopted by vaccinia virus (vv) to evade the host immune system is to encode homologs of tnf receptors (tnfrs) that block tnf-α function. the response to vv skin infection under conditions of tnf-α deficiency, however, has not been reported. we found that tnfr1-/- mice developed larger primary lesions, numerous satellite lesions, and higher skin virus levels after vv scarification. following their recovery, vv-scarified tnfr1-/- mice were fully protected against challenge with a leth ...201222318381
superior induction of t cell responses to conserved hiv-1 regions by electroporated alphavirus replicon dna compared to that with conventional plasmid dna vaccine.vaccination using "naked" dna is a highly attractive strategy for induction of pathogen-specific immune responses; however, it has been only weakly immunogenic in humans. previously, we constructed dna-launched semliki forest virus replicons (drep), which stimulate pattern recognition receptors and induce augmented immune responses. also, in vivo electroporation was shown to enhance immune responses induced by conventional dna vaccines. here, we combine these two approaches and show that in vivo ...201222318135
oncolytic virus and anti-4-1bb combination therapy elicits strong antitumor immunity against established cancer.oncolytic virotherapy using vaccinia virus (vv) has shown some encouraging antitumor responses in mouse models and patients, but the breadth of efficacy in clinical trials has been somewhat limited. given that antitumor effects have correlated with increased host immune responses, we hypothesized that improved therapeutic outcomes may be achieved by using oncolytic virus (ov) in combination with a potent immune agonist reagent. in this study, we carried out a preclinical evaluation of a genetica ...201222315352
oncolytic virotherapy for hematological malignancies.hematological malignancies such as leukemias, lymphomas, multiple myeloma (mm), and the myelodysplastic syndromes (mdss) primarily affect adults and are difficult to treat. for high-risk disease, hematopoietic stem cell transplant (hct) can be used. however, in the setting of autologous hct, relapse due to contamination of the autograft with cancer cells remains a major challenge. ex vivo manipulations of the autograft to purge cancer cells using chemotherapies and toxins have been attempted. be ...201122312362
viral infection triggers rapid differentiation of human blood monocytes into dendritic cells.surprisingly little is known about the interaction of human blood mononuclear cells with viruses. here, we show that monocytes are the predominant cell type infected when peripheral blood mononuclear cells are exposed to viruses ex vivo. remarkably, infection with vesicular stomatitis virus, vaccinia virus, and a variety of influenza a viruses (including circulating swine-origin virus) induces monocytes to differentiate within 18 hours into cd16(-)cd83(+) mature dendritic cells with enhanced cap ...201222310910
aptamer-based viability impedimetric sensor for viruses.the development of aptamer-based viability impedimetric sensor for viruses (aptavisens-v) is presented. highly specific dna aptamers to intact vaccinia virus were selected using cell-selex technique and integrated into impedimetric sensors via self-assembly onto a gold microelectrode. remarkably, this aptasensor is highly selective and can successfully detect viable vaccinia virus particles (down to 60 virions in a microliter) and distinguish them from nonviable viruses in a label-free electroch ...201222303883
the mc159 protein from the molluscum contagiosum poxvirus inhibits nf-κb activation by interacting with the iκb kinase complex.molluscum contagiosum virus (mcv) causes persistent neoplasms in healthy and immunocompromised people. its ability to persist likely is due to its arsenal of viral immunoevasion proteins. for example, the mcv mc159 protein inhibits tnf-r1-induced nf-κb activation and apoptosis. the mc159 protein is a viral flip and, as such, possesses two tandem death effector domains (deds). we show in this article that, in human embryonic kidney 293 t cells, the expression of wild-type mc159 or a mutant mc159 ...201222301546
the difference in il-1beta , mip-1alpha, il-8 and il-18 production between the infection of pma activated u937 cells with recombinant vaccinia viruses inserted 2004 h5n1 influenza ha genes and ns genes.the severity of avian influenza h5n1 disease is correlated with the ability of the virus to induce an over production of proinflammatory cytokines from innate immune cells. however, the role of each virus gene is unknown. to elaborate the function of each virus gene, the recombinant vaccinia virus inserted ha and ns gene from the 2004 h5n1 virus were used in the study.201122299315
role of metalloproteases in vaccinia virus epitope processing for transporter associated with antigen processing (tap)-independent human leukocyte antigen (hla)-b7 class i antigen presentation.the transporter associated with antigen processing (tap) translocates the viral proteolytic peptides generated by the proteasome and other proteases in the cytosol to the endoplasmic reticulum lumen. there, they complex with nascent human leukocyte antigen (hla) class i molecules, which are subsequently recognized by the cd8(+) lymphocyte cellular response. however, individuals with nonfunctional tap complexes or tumor or infected cells with blocked tap molecules are able to present hla class i ...201222298786
eczema vaccinatum.eczema vaccinatum (ev) is a complication of smallpox vaccination that can occur in persons with eczema/atopic dermatitis (ad), in which vaccinia virus disseminates to cause an extensive rash and systemic illness. because persons with eczema are deferred from vaccination, only a single, accidentally transmitted case of ev has been described in the medical literature since military vaccination was resumed in the united states in 2002. to enhance understanding of ev, we review its history during th ...201222291103
susceptibility to vaccinia virus infection and spread in mice is determined by age at infection, allergen sensitization and mast cell status.patients, especially young children, with atopic dermatitis are at an increased risk of developing eczema vaccinatum, a severe reaction to the smallpox vaccine, either through direct vaccination or indirect contact with a person recently vaccinated.201222286752
recruitment of host translation initiation factor eif4g by the vaccinia virus ssdna-binding protein i3.poxviruses are large double-stranded dna viruses that replicate exclusively in the cytoplasm of infected cells within discrete compartments termed viral factories. recent work has shown that the prototypical poxvirus, vaccinia virus (vacv) sequesters components of the eukaryotic translation initiation complex eif4f within viral factories while also stimulating formation of eif4f complexes. however, the forces that govern these events remain unknown. here, we show that maximal eif4f formation req ...201222280895
glomulin: a permissivity factor for vaccinia virus infection.in earlier studies we provided evidence that vaccinia virus (vacv) phosphorylation-activation of host cell signaling effectors is critical for subsequent viral replication. in this report, using mass spectrometry-based proteomics, we have identified 387 host cell proteins that co-immunoprecipitate with vacv in infected, permissive pm1.ccr5 human t cells. among these, glomulin was distinguishable based on its known interaction with a tyrosine kinase receptor, c-met, its ability to become tyrosine ...201222280104
vaccinia mature virus fusion regulator a26 protein binds to a16 and g9 proteins of the viral entry fusion complex and dissociates from mature virions at low ph.vaccinia mature virus enters cells through either endocytosis or plasma membrane fusion, depending on virus strain and cell type. our previous results showed that vaccinia virus mature virions containing viral a26 protein enter hela cells preferentially through endocytosis, whereas mature virions lacking a26 protein enter through plasma membrane fusion, leading us to propose that a26 acts as an acid-sensitive fusion suppressor for mature virus (s. j. chang, y. x. chang, r. izmailyan r, y. l. tan ...201222278246
species specificity of protein kinase r antagonism by cytomegalovirus trs1 genes.the host antiviral protein kinase r (pkr) has rapidly evolved during primate evolution, likely in response to challenges posed by many different viral antagonists, such as the trs1 gene of cytomegaloviruses (cmvs). in turn, viral antagonists have adapted to changes in pkr. as a result of this "arms race," modern trs1 alleles in cmvs may function differently in cells derived from alternative species. we have previously shown that human cmv trs1 (hutrs1) blocks the pkr pathway and rescues replicat ...201222278235
clinical assessment of a recombinant simian adenovirus chad63: a potent new vaccine vector.background. vaccine development in human plasmodium falciparum malaria has been hampered by the exceptionally high levels of cd8(+) t cells required for efficacy. use of potently immunogenic human adenoviruses as vaccine vectors could overcome this problem, but these are limited by preexisting immunity to human adenoviruses.methods. from 2007 to 2010, we undertook a phase i dose and route finding study of a new malaria vaccine, a replication-incompetent chimpanzee adenovirus 63 (chad63) encoding ...201222275401
a new stable and reliable method for labeling nucleic acids of fully replicative viruses.efficiently labeling nucleic acids of fully replicative viruses is a challenge. in this work, a 'molecular light switch' complex [ru(phen)(2)(dppz)](2+), where phen = 1,10-phenanthroline and dppz = dipyrido[3,2-a:2',3'-c]phenazine, has been exploringly used to label vaccinia virus nucleic acid. the labeled virions exhibited strong and stable fluorescence and could be imaged at the single-virion level. moreover, they were fully infectious and can be used to study the behaviors of invasion into th ...201222273842
ox40 ligand and programmed cell death 1 ligand 2 expression on inflammatory dendritic cells regulates cd4 t cell cytokine production in the lung during viral disease.cd4 th differentiation is influenced by costimulatory molecules expressed on conventional dendritic cells (dcs) in regional lymph nodes and results in specific patterns of cytokine production. however, the function of costimulatory molecules on inflammatory (cd11b(+)) dcs in the lung during recall responses is not fully understood, but it is important for development of novel interventions to limit immunopathological responses to infection. using a mouse model in which vaccination with vaccinia ...201222266281
vaccinia virus zoonotic infection, são paulo state, brazil. 201222260819
a novel genetically modified oncolytic vaccinia virus in experimental models is effective against a wide range of human cancers.background: replication-competent oncolytic viruses have shown great promise as a potential cancer treatment. this study aimed to determine whether a novel vaccinia virus, glv-1h151, with genetic modifications enhancing cancer specificity and enabling virus detection, is effective against a range of human cancers and is safe when administered in preclinical models. methods: glv-1h151 was modified with deletion of ...201222258815
failure of smallpox vaccine to take in vaccinia-naive individuals is related to differences in antibody profiles before vaccination, not after.successful vaccination against smallpox with conventional vaccinia virus is usually determined by the development of a vesicular skin lesion at the site of vaccinia inoculation, called a 'take'. although previous vaccination is known to be associated with attenuation of the take, the immunology that underlies a no-take in vaccinia-naïve individuals is not well understood. we hypothesized that antibody profiling of individuals before and after receiving vaccinia would reveal differences between t ...201222258709
coated microneedle arrays for transcutaneous delivery of live virus vaccines.vaccines are sensitive biologics that require continuous refrigerated storage to maintain their viability. the vast majority of vaccines are also administered using needles and syringes. the need for cold chain storage and the significant logistics surrounding needle-and-syringe vaccination is constraining the success of immunization programs. recombinant live viral vectors are a promising platform for the development of vaccines against a number of infectious diseases, however these viruses mus ...201122245683
vaccinia viruses isolated from skin infection in horses produced cutaneous and systemic disease in experimentally infected rabbits.the susceptibility of rabbits to two isolates of vaccinia virus (vacv) recovered from cutaneous disease in horses in southern brazil was investigated. rabbits were inoculated in the ear skin with both vacv isolates, either in single or mixed infection. all inoculated animals presented local skin lesions characterized by hyperaemia, papules, vesicles, pustules and ulcers. infectious virus was detected in the lungs and intestine of rabbits that died during acute disease. histological examination o ...201222244689
mutagenesis of the palmitoylation site in vaccinia virus envelope glycoprotein b5.the outer envelope of vaccinia virus extracellular virions is derived from intracellular membranes that, at late times in infection, are enriched in several virus-encoded proteins. although palmitoylation is common in vaccinia virus envelope proteins, little is known about the role of palmitoylation in the biogenesis of the enveloped virus. we have studied the palmitoylation of b5, a 42 kda type i transmembrane glycoprotein comprised of a large ectodomain and a short (17 aa) cytoplasmic tail. mu ...201222238237
cyclophilin a is required for efficient human cytomegalovirus dna replication and reactivation.human cytomegalovirus (hcmv) is a large dna virus belonging to the subfamily betaherpesvirinae. haematopoietic cells of the myeloid lineage have been shown to harbour latent hcmv. however, following terminal differentiation of these cells, virus is reactivated, and in an immunocompromised host acute infection can occur. it is currently unknown which viral and cellular factors are involved in regulating the switch between lytic and latent infections. cyclophilin a (cypa) is a cellular protein tha ...201222238232
functional hyper-il-6 from vaccinia virus-colonized tumors triggers platelet formation and helps to alleviate toxicity of mitomycin c enhanced virus therapy.abstract: background: combination of oncolytic vaccinia virus therapy with conventional chemotherapy has shown promise for tumor therapy. however, side effects of chemotherapy including thrombocytopenia, still remain problematic. methods: here, we describe a novel approach to optimize combination therapy of oncolytic virus and chemotherapy utilizing virus-encoding hyper il-6, glv-1h90, to reduce chemotherapy-associated side effects. results: we showed that the hyper-il-6 cytokine was successful ...201222236378
immune modulation in primary vaccinia virus zoonotic human infections.in 2010, the who celebrated the 30th anniversary of the smallpox eradication. ironically, infections caused by viruses related to smallpox are being increasingly reported worldwide, including monkeypox, cowpox, and vaccinia virus (vacv). little is known about the human immunological responses elicited during acute infections caused by orthopoxviruses. we have followed vacv zoonotic outbreaks taking place in brazil and analyzed cellular immune responses in patients acutely infected by vacv. resul ...201122229039
vaccinia virus-induced smallpox postvaccinal encephalitis in case of blood-brain barrier damage.smallpox vaccination is the only currently effective mean to combat the threat of variola virus used as a bioterrorism agent, although it is responsible for a rare but serious complication, the postvaccinal encephalitis (pve). development of safer vaccines therefore is a high priority as the pve physiopathology is not well understood to date. if vaccinia virus (vacv) is responsible for pve by central nervous system (cns) dissemination, trans-migration of the vacv across the blood-brain barrier ( ...201222227123
in vitro inhibition of monkeypox virus production and spread by interferon-β.the orthopoxvirus genus contains numerous virus species that are capable of causing disease in humans, including variola virus (the etiological agent of smallpox), monkeypox virus, cowpox virus, and vaccinia virus (the prototypical member of the genus). monkeypox is a zoonotic disease that is endemic in the democratic republic of the congo and is characterized by systemic lesion development and prominent lymphadenopathy. like variola virus, monkeypox virus is a high priority pathogen for therape ...201222225589
cutting edge: protective effect of cx3cr1+ dendritic cells in a vaccinia virus pulmonary infection model.the protective host immune response to viral infections requires both effective innate and adaptive immune responses. cross-talk between the two responses is coordinated by the chemokine network and professional apcs such as dendritic cells (dcs). in mice, subpopulations of myeloid dcs in peripheral tissues such as lungs and in blood express cx3cr1 depending on the inflammation state. we thus examined the host response of mice deficient in the chemokine receptor cx3cr1 to an intranasal vaccinia ...201222219332
oncolytic virotherapy in veterinary medicine: current status and future prospects for canine patients.oncolytic viruses refer to those that are able to eliminate malignancies by direct targeting and lysis of cancer cells, leaving non-cancerous tissues unharmed. several oncolytic viruses including adenovirus strains, canine distemper virus and vaccinia virus strains have been used for canine cancer therapy in preclinical studies. however, in contrast to human studies, clinical trials with oncolytic viruses for canine cancer patients have not been reported. an 'ideal' virus has yet to be identifie ...201222216938
a viral vectored prime-boost immunization regime targeting the malaria pfs25 antigen induces transmission-blocking activity.the ookinete surface protein pfs25 is a macrogamete-to-ookinete/ookinete stage antigen of plasmodium falciparum, capable of exerting high-level anti-malarial transmission-blocking activity following immunization with recombinant protein-in-adjuvant formulations. here, this antigen was expressed in recombinant chimpanzee adenovirus 63 (chad63), human adenovirus serotype 5 (adhu5) and modified vaccinia virus ankara (mva) viral vectored vaccines. two immunizations were administered to mice in a het ...201122216279
cross-presentation and genome-wide screening reveal candidate t cells antigens for a herpes simplex virus type 1 vaccine.herpes simplex virus type 1 (hsv-1) not only causes painful recurrent oral-labial infections, it can also cause permanent brain damage and blindness. there is currently no hsv-1 vaccine. an effective vaccine must stimulate coordinated t cell responses, but the large size of the genome and the low frequency of hsv-1-specific t cells have hampered the search for the most effective t cell antigens for inclusion in a candidate vaccine. we have now developed what we believe to be novel methods to eff ...201222214845
Casein kinase 2 regulates vaccinia virus actin tail formation.Casein kinase 2 (CK2) is a pleiotropic serine/threonine kinase that regulates numerous cellular processes and is essential to the infectious cycle of several viruses. Here we investigated the potential role of CK2 in vaccinia virus (VACV) infection. We used the CK2 inhibitor TBB and found that CK2 inactivation impaired VACV dissemination and actin tail formation. We used RNAi and confirmed that CK2 depletion impaired VACV actin tail formation. Furthermore, we designed a recombinant virus that al ...201122209233
sex differences in murine susceptibility to systemic viral infections.increased susceptibility to autoimmunity in females is often viewed as the consequence of enhanced immunoreactivity providing superior protection against infections. we paradoxically observed greater mortality in female compared to male mice during systemic viral infections with three large double-stranded dna viruses (herpes simplex virus type i [hsv], murine cytomegalovirus [mcmv], and vaccinia virus [vv]). indeed, female mice were 27-fold more susceptible to infection with hsv than male mice. ...201122209097
identification of a pyridopyrimidinone inhibitor of orthopoxviruses from a diversity-oriented synthesis library.orthopoxviruses include the prototypical vaccinia virus, the emerging infectious agent monkeypox, and potential biothreat variola (smallpox). there is currently no fda-approved treatment for humans infected with orthopoxviruses. we screened a diversity oriented synthesis library for new scaffolds with antiviral activity against vaccinia. this screen identified a non-nucleoside analog that blocked post-replicative intermediate and late gene expression. viral genome replication was unaffected, and ...201122205744
Distinct Humoral and Cellular Immunity Induced by Alternating Prime-boost Vaccination Using Plasmid DNA and Live Viral Vector Vaccines Expressing the E Protein of Dengue Virus Type 2.Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2).201122194710
Granzyme B Inhibits Vaccinia Virus Production through Proteolytic Cleavage of Eukaryotic Initiation Factor 4 Gamma 3.Cytotoxic T lymphocytes (CTLs) are the major killer of virus-infected cells. Granzyme B (GrB) from CTLs induces apoptosis in target cells by cleavage and activation of substrates like caspase-3 and Bid. However, while undergoing apoptosis, cells are still capable of producing infectious viruses unless a mechanism exists to specifically inhibit viral production. Using proteomic approaches, we identified a novel GrB target that plays a major role in protein synthesis: eukaryotic initiation factor ...201122194691
The membrane fusion step of vaccinia virus entry is cooperatively mediated by multiple viral proteins and host cell components.For many viruses, one or two proteins allow cell attachment and entry, which occurs through the plasma membrane or following endocytosis at low pH. In contrast, vaccinia virus (VACV) enters cells by both neutral and low pH routes; four proteins mediate cell attachment and twelve that are associated in a membrane complex and conserved in all poxviruses are dedicated to entry. The aim of the present study was to determine the roles of cellular and viral proteins in initial stages of entry, specifi ...201122194690
inhibition of apoptosis and nf-κb activation by vaccinia protein n1 occur via distinct binding surfaces and make different contributions to virulence.vaccinia virus (vacv) protein n1 is an intracellular virulence factor and belongs to a family of vacv b-cell lymphoma (bcl)-2-like proteins whose members inhibit apoptosis or activation of pro-inflammatory transcription factors, such as interferon (ifn) regulatory factor-3 (irf-3) and nuclear factor-κb (nf-κb). unusually, n1 inhibits both apoptosis and nf-κb activation. to understand how n1 exerts these different functions, we have mutated residues in the bcl-2-like surface groove and at the int ...201122194685
contact transmission of vaccinia virus from smallpox vaccinees in the united states, 2003-2011.since 2002, approximately 40,000 us civilians and 2.1 million military personnel have been vaccinated against smallpox. the vaccine contains live vaccinia virus that can be transferred through physical contact. this report summarizes numbers, rates, and characteristics of contact vaccinia cases that presented between december 2002 and march 2011. cases were identified from reports in adverse event reporting systems and peer-reviewed literature. one hundred fifteen cases of vaccinia transmission ...201122192851
Generation of recombinant Orf virus using an enhanced green fluorescent protein reporter gene as a selectable marker.ABSTRACT: BACKGROUND: Reporter genes are often used as a selectable marker for generation of recombinant viruses in order to investigate the mechanism of pathogenesis and to obtain candidate vaccine viruses. Routine selection of the recombinant parapoxvirus is time-consuming and labor intensive. Therefore, developing a novel method for selection is critical. RESULTS: In this study, we developed a rapid method to generate recombinant Orf viruses (ORFV) based on the enhanced green fluorescent p ...201122192523
the oncolytic poxvirus jx-594 selectively replicates in and destroys cancer cells driven by genetic pathways commonly activated in cancers.oncolytic viruses are generally designed to be cancer selective on the basis of a single genetic mutation. jx-594 is a thymidine kinase (tk) gene-inactivated oncolytic vaccinia virus expressing granulocyte-macrophage colony-stimulating factor (gm-csf) and lac-z transgenes that is designed to destroy cancer cells through replication-dependent cell lysis and stimulation of antitumoral immunity. jx-594 has demonstrated a favorable safety profile and reproducible tumor necrosis in a variety of solid ...201122186794
inhibition of cowpox virus and monkeypox virus infection by mitoxantrone.mitoxantrone, an fda-approved therapeutic for the treatment of cancer and multiple sclerosis, was previously reported to exhibit antiviral activity against vaccinia virus. to determine whether this activity extends to other orthopoxviruses, mitoxantrone was tested against cowpox and monkeypox. mitoxantrone demonstrated an ec(50) of 0.25μm against cowpox and 0.8μm against monkeypox. intraperitoneal treatment of cowpox virus-challenged c57bl/6 mice with 0.5mg/kg mitoxantrone resulted in 25% surviv ...201122182595
Cryo X-ray nano-tomography of vaccinia virus infected cells.We have performed full-field cryo X-ray microscopy in the water window photon energy range on vaccinia virus (VACV) infected cells to produce tomographic reconstructions. PtK2 cells were infected with a GFP-expressing VACV strain and frozen by plunge fast freezing. The infected cells were selected by light fluorescence microscopy of the GFP marker and subsequently imaged in the X-ray microscope under cryogenic conditions. Tomographic tilt series of X-ray images were used to yield three-dimension ...201122178221
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