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the vaccinia virus k7 protein promotes histone methylation associated with heterochromatin formation.it has been well established that many vaccinia virus proteins suppress host antiviral pathways by targeting the transcription of antiviral proteins, thus evading the host innate immune system. however, whether viral proteins have an effect on the host's overall cellular transcription is less understood. in this study we investigated the regulation of heterochromatin during vaccinia virus infection. heterochromatin is a highly condensed form of chromatin that is less transcriptionally active and ...201728257484
survival of tissue-resident memory t cells requires exogenous lipid uptake and metabolism.tissue-resident memory t (trm) cells persist indefinitely in epithelial barrier tissues and protect the host against pathogens. however, the biological pathways that enable the long-term survival of trm cells are obscure. here we show that mouse cd8(+) trm cells generated by viral infection of the skin differentially express high levels of several molecules that mediate lipid uptake and intracellular transport, including fatty-acid-binding proteins 4 and 5 (fabp4 and fabp5). we further show that ...201728219080
divergent roles of β- and γ-actin isoforms during spread of vaccinia virus.actin is a major component of the cytoskeleton and is present as two isoforms in non-muscle cells: β- and γ-cytoplasmic actin. these isoforms are strikingly conserved, differing by only four n-terminal amino acids. during spread from infected cells, vaccinia virus (vacv) particles induce localized actin nucleation that propel virus to surrounding cells and facilitate cell-to-cell spread of infection. here we show that virus-tipped actin comets are composed of β- and γ-actin. we employed isoform- ...201728218453
ac dielectrophoretic manipulation and electroporation of vaccinia virus using carbon nanoelectrode arrays.this paper reports the capture and detection of vaccinia virus particles based on ac dielectrophoresis (dep) and electrochemical impedance measurements employing an embedded vertically aligned carbon nanofiber (vacnf) nanoelectrode array (nea) versus a macroscopic indium-tin-oxide (ito) transparent electrode in a "points-and-lid" configuration. the nano-dep device was fabricated by bonding two su-8 covered electrodes patterned using photolithography. the bottom electrode contains a 200 × 200 μm( ...201728211116
establishing elements of a synthetic biology platform for vaccinia virus production: biobrick™ design, serum-free virus production and microcarrier-based cultivation of cv-1 cells.vaccinia virus (vacv) is an established vector for vaccination and is beginning to prove effective as an oncolytic agent. industrial production of vacv stands to benefit in future from advances made by synthetic biology in genome engineering and standardisation. the cv-1 cell line can be used for vacv propagation and has been used extensively with the crispr/cas9 system for making precise edits of the vacv genome. here we take first steps toward establishing a scalable synthetic biology platform ...201728203643
proteomic screen for cellular targets of the vaccinia virus f10 protein kinase reveals that phosphorylation of mdia regulates stress fiber formation.vaccinia virus, a complex dsdna virus, is unusual in replicating exclusively within the cytoplasm of infected cells. although this prototypic poxvirus encodes >200 proteins utilized during infection, a significant role for host proteins and cellular architecture is increasingly evident. the viral b1 kinase and h1 phosphatase are known to target cellular proteins as well as viral substrates, but little is known about the cellular substrates of the f10 kinase. f10 is essential for virion morphogen ...201728183815
investigating viruses during the transformation of molecular biology.this reflections article describes my early work on viral enzymes and the discovery of mrna capping, how my training in medicine and biochemistry merged as i evolved into a virologist, the development of viruses as vaccine vectors, and how scientific and technological developments during the 1970s and beyond set the stage for the interrogation of nearly every step in the reproductive cycle of vaccinia virus (vacv), a large dna virus with about 200 genes. the reader may view this article as a wor ...201728154190
entirely plasmid-based reverse genetics system for rotaviruses.rotaviruses (rvs) are highly important pathogens that cause severe diarrhea among infants and young children worldwide. the understanding of the molecular mechanisms underlying rv replication and pathogenesis has been hampered by the lack of an entirely plasmid-based reverse genetics system. in this study, we describe the recovery of recombinant rvs entirely from cloned cdnas. the strategy requires coexpression of a small transmembrane protein that accelerates cell-to-cell fusion and vaccinia vi ...201728137864
online size-exclusion and ion-exchange chromatography on a saxs beamline.biological small angle x-ray scattering (biosaxs) is a powerful technique in molecular and structural biology used to determine solution structure, particle size and shape, and surface-to-volume ratio of macromolecules. the technique is applicable to a very wide variety of solution conditions spanning a broad range of concentrations, ph values, ionic strengths, temperatures, additives, etc., but the sample is required to be monodisperse. this caveat led to the implementation of liquid chromatogr ...201728117806
detection of vaccinia virus in urban domestic cats, brazil.we investigated possible vaccinia virus (vacv) in urban house cats in brazil. serum samples from 6 cats were positive for vacv by pcr, indicating likely vacv circulation among house cats in urban areas of brazil. this finding highlights the importance of epidemiologic surveillance to avoid outbreaks among urban human populations.201728098542
coherent brightfield microscopy provides the spatiotemporal resolution to study early stage viral infection in live cells.viral infection starts with a virus particle landing on a cell surface followed by penetration of the plasma membrane. due to the difficulty of measuring the rapid motion of small-sized virus particles on the membrane, little is known about how a virus particle reaches an endocytic site after landing at a random location. here, we use coherent brightfield (cobri) microscopy to investigate early stage viral infection with ultrahigh spatiotemporal resolution. by detecting intrinsic scattered light ...201728067508
infectio: a generic framework for computational simulation of virus transmission between cells.viruses spread between cells, tissues, and organisms by cell-free and cell-cell mechanisms, depending on the cell type, the nature of the virus, or the phase of the infection cycle. the mode of viral transmission has a large impact on disease development, the outcome of antiviral therapies or the efficacy of gene therapy protocols. the transmission mode of viruses can be addressed in tissue culture systems using live-cell imaging. yet even in relatively simple cell cultures, the mechanisms of vi ...201727303704
viruses as nanomedicine for cancer.oncolytic virotherapy, a type of nanomedicine in which oncolytic viruses (ovs) are used to selectively infect and lyse cancer cells, is an emerging field in cancer therapy. some ovs exhibit a specific tropism for cancer cells, whereas others require genetic modification to enhance their binding with and entry into cancer cells. ovs both kill tumor cells and induce the host's immune response against tumor cells. armed with antitumor cellular molecules, antibodies, and/or in combination with antic ...201727703350
a novel anti-viral role for stat3 in ifn-α signalling responses.the cytokine, interferon (ifn)-α, induces a wide spectrum of anti-viral mediators, via the janus kinase/signal transducer and activator of transcription (jak/stat) pathway. stat1 and stat2 are well characterised to upregulate ifn-stimulated gene (isg) expression; but even though stat3 is also activated by ifn-α, its role in anti-viral isg induction is unclear. several viruses, including hepatitis c and mumps, reduce cellular stat3 protein levels, via the promotion of ubiquitin-mediated proteasom ...201727988795
molecular evidence of orthopoxvirus dna in capybara (hydrochoerus hydrochaeris) stool samples.vaccinia virus (vacv) is responsible for outbreaks in brazil and has immense potential as an emerging virus. vacv can be found naturally circulating in india, pakistan and south america, where it causes infections characterised by exanthematic lesions in buffaloes, cattle and humans. the transmission cycle of brazilian vacv has still not been fully characterised; one of the most important gaps in knowledge being the role of wild animals. capybaras, which are restricted to the americas, are the w ...201727771792
identification and functional analysis of the gtpv bidirectional promoter region.the goat pox chick embryo-attenuated virus (gtpv) has been developed as an effective vaccine that can elicit protective immune responses. it possesses a large genome and a robust ability to express exogenous genes. thus, this virus is an ideal vector for recombinant live vaccines for infectious diseases in ruminant animals. in this study, we identified a novel bidirectional promoter region of gtpv through screening named pbvv(±). pbvv(±) is located between etf-l and vitf-3, which are transcribed ...201727771746
ox40 cooperates with icos to amplify follicular th cell development and germinal center reactions during infection.cognate interactions between t follicular helper (tfh) cells and b cells are essential for promoting protective ab responses. whereas costimulatory receptors such as icos are accepted as being important for the induction of tfh cell fate decision, other molecules may play key roles in amplifying or maintaining the tfh phenotype. in this study, with vaccinia virus infection in mice, we show that ox40 was expressed on tfh cells that accumulated at the t/b borders in the white pulp of the spleen an ...201727895177
detection of vaccinia virus during an outbreak of exanthemous oral lesions in brazilian equids.in august 2014, an outbreak of oral exanthematous disease in equids was reported in brazil, affecting 11 donkeys and 3 mules.201726875684
enhancing expression of functional human sodium iodide symporter and somatostatin receptor in recombinant oncolytic vaccinia virus for in vivo imaging of tumors.oncolytic virus (ov) therapy has emerged as a novel tool in our therapeutic arsenals for fighting cancer. as a live biologic agent, ov has the ability to target and selectively amplify at the tumor sites. we have reported that a vaccinia-based ov (pexa-vec) has shown good efficacy in preclinical models and in clinical trials. to give an additional tool to clinicians to allow both treatment of the tumor and improved visualization of tumor margins, we developed new viral-based platforms with 2 spe ...201727635026
development of recombinant canarypox viruses expressing immunogens.canarypox viruses (cnpv) are excellent candidates to develop recombinant vector vaccines due to both their capability to induce protective immune responses and their incompetence to replicate in mammalian cells (safety profile). in addition, cnpv and the derived recombinants can be manipulated under biosafety level 1 conditions. there is no commercially available system to obtain recombinant cnpv; however, the methodology and tools required to develop recombinant vaccinia virus (vv), prototype o ...201728374241
virus inactivation under the photodynamic effect of phthalocyanine zinc(ii) complexes.various metal phthalocyanines have been studied for their capacity for photodynamic effects on viruses. two newly synthesized water-soluble phthalocyanine zn(ii) complexes with different charges, cationic methylpyridyloxy-substituted zn(ii)- phthalocyanine (znpcme) and anionic sulfophenoxy-substituted zn(ii)-phthalocyanine (znpcs), were used for photoinactivation of two dna-containing enveloped viruses (herpes simplex virus type 1 and vaccinia virus), two rna-containing enveloped viruses (bovine ...201727845890
a multiplex pcr for viruses associated with exanthematic and vesicular disease in cattle.exanthematic and papulo-vesicular lesions in the udder and teats of milking cows are fairly common in some brazilian dairies, especially those with poor sanitary conditions and hand milking. the orthopoxvirus vaccinia virus (vacv) and the parapoxviruses pseudocowpox virus (pcpv) and bovine popular stomatitis virus (bpsv) have been frequently associated with such conditions. elsewhere, bovine herpesvirus 2 (bohv-2) has also been associated with similar clinical signs. thus, we herein describe a c ...201727793645
antiviral activity and metal ion-binding properties of some 2-hydroxy-3-methoxyphenyl acylhydrazones.here we report on the results obtained from an antiviral screening, including herpes simplex virus, vaccinia virus, vesicular stomatitis virus, coxsackie b4 virus or respiratory syncytial virus, parainfluenza-3 virus, reovirus-1 and punta toro virus, of three 2-hydroxy-3-methoxyphenyl acylhydrazone compounds in three cell lines (i.e. human embryonic lung fibroblast cells, human cervix carcinoma cells, and african green monkey kidney cells). interesting antiviral ec50 values are obtained against ...201829209895
maternal hiv-1 env vaccination for systemic and breast milk immunity to prevent oral shiv acquisition in infant macaques.mother-to-child transmission (mtct) of human immunodeficiency virus type 1 (hiv-1) contributes to an estimated 150,000 new infections annually. maternal vaccination has proven safe and effective at mitigating the impact of other neonatal pathogens and is one avenue toward generating the potentially protective immune responses necessary to inhibit hiv-1 infection of infants through breastfeeding. in the present study, we tested the efficacy of a maternal vaccine regimen consisting of a modified v ...201829359183
vaccinia virus in feces and urine of wild rodents from são paulo state, brazil.the origin of vaccinia virus (vacv) outbreaks in brazil remains unknown, but since the isolation of vacv in mus musculus mice during a zoonotic outbreak affecting cattle and milkers, peridomestic rodents have been suggested to be a link between cows and wild animals. considering that experimentally infected mice eliminate viral particles in their feces, we investigated the presence of vacv in the feces and urine of wild rodents that were captured in the forest areas surrounding milking farms in ...201829360742
vaccinia virus in blood samples of humans, domestic and wild mammals in brazil.outbreaks of vaccinia virus (vacv) affecting cattle and humans have been reported in brazil in the last 15 years, but the origin of outbreaks remains unknown. although vacv dna have been already detected in mice (mus musculus), opossums (didelphis albiventris) and dogs during vacv zoonotic outbreaks, no transmission to cattle or humans from any of these were reported during brazilian outbreaks. in this work, we assessed the pcr positivity to vacv in blood samples of cows and other domestic mamma ...201829346277
bovine vaccinia: insights into the disease in cattle.bovine vaccinia (bv), caused byvaccinia virus(vacv), is a zoonosis characterized by exanthematous lesions in the teats of dairy cows and the hands of milkers and is an important public health issue. severe vacv-induced lesions in the teats and udder of cows and buffaloes could lead to mastitis and other secondary infections, thereby reducing productivity and resulting in economic losses to the dairy industry. in brazil, bv re-emerged in the late 1990s and is now endemic in most of the brazilian ...201829522489
prime and boost vaccination elicit a distinct innate myeloid cell immune response.understanding the innate immune response to vaccination is critical in vaccine design. here, we studied blood innate myeloid cells after first and second immunization of cynomolgus macaques with the modified vaccinia virus ankara. the inflammation at the injection site was moderate and resolved faster after the boost. the blood concentration of inflammation markers increased after both injections but was lower after the boost. the numbers of neutrophils, monocytes, and dendritic cells were trans ...201829449630
buffalopox.buffalopox is a contagious viral disease affecting milch buffaloes (bubalus bubalis) and, rarely, cows. the disease has zoonotic implications, as outbreaks are frequently associated with human infections, particularly in the milkers. buffalopox is associated with high morbidity (80%). the clinical symptoms of the disease are characterized by wartline lesions on the udder, teats, inguinal region, base of the ears, and over the parotid. in the severe form, generalized rash is observed. although th ...201829323851
the immune response of rhesus macaques to novel vaccines comprising hepatitis b virus s, pres1, and core antigens.therapeutic vaccines represent a unique approach to hepatitis b virus (hbv) treatment and have the potential to induce long-term control of infection. this study explored the immune responses of rhesus macaques to novel vaccines comprising the s, pres1, and core antigens of the hbv that showed promise as prophylactic and therapeutic approaches in a mouse model. the tested vaccines included two dna vaccines (pvrc-ss1, pvrc-cs1), an hbv particle subunit (hbss1) vaccine and the recombinant vaccinia ...201829778513
dynamic transcriptome profiling dataset of vaccinia virus obtained from long-read sequencing techniques.poxviruses are large dna viruses that infect humans and animals. vaccinia virus (vacv) has been applied as a live vaccine for immunization against smallpox, which was eradicated by 1980 as a result of worldwide vaccination. vacv is the prototype of poxviruses in the investigation of the molecular pathogenesis of the virus. short-read sequencing methods have revolutionized transcriptomics; however, they are not efficient in distinguishing between the rna isoforms and transcript overlaps. long-rea ...201830476066
short communication: parapoxvirus and orthopoxvirus coinfection in milk of naturally infected cows.several studies have shown the occurrence of poxvirus infections associated with exanthematic lesions in cattle from many brazilian states. coinfection between viruses belonging to 2 genera, orthopoxvirus (opxv) and parapoxvirus (ppv), was already identified from the lesions of affected cows and humans. the dna and infectious viral particles of vaccinia virus, an opxv, have been detected in milk of naturally and experimentally infected cows. however, to date no reports have described the detecti ...201830007812
experimental infection of dromedaries with middle east respiratory syndrome-coronavirus is accompanied by massive ciliary loss and depletion of the cell surface receptor dipeptidyl peptidase 4.middle east respiratory syndrome (mers) represents an important respiratory disease accompanied by lethal outcome in one-third of human patients. recent data indicate that dromedaries represent an important source of infection, although information regarding viral cell tropism and pathogenesis is sparse. in the current study, tissues of eight dromedaries receiving inoculation of mers-coronavirus (mers-cov) after recombinant modified-vaccinia-virus-ankara (mva-s)-vaccination (n = 4), mva-vaccinat ...201829950581
tracking modified vaccinia virus ankara in the chicken embryo: in vivo tropism and pathogenesis of egg infections.the modified vaccinia virus ankara (mva) is a highly attenuated vaccinia virus serving as a promising vector vaccine platform to develop vaccines against infectious diseases. in contrast to the well-established replication deficiency and safety of mva in mammals, much less is known about mva infection in avian hosts. here, we used a recombinant mva expressing fluorescent reporter proteins under transcriptional control of specific viral early and late promoters to study in vivo tropism, distribut ...201830149505
removal of the c6 vaccinia virus interferon-β inhibitor in the hepatitis c vaccine candidate mva-hcv elicited in mice high immunogenicity in spite of reduced host gene expression.hepatitis c virus (hcv) represents a major global health problem for which a vaccine is not available. modified vaccinia virus ankara (mva)-hcv is a unique hcv vaccine candidate based in the modified vaccinia virus ankara (mva) vector expressing the nearly full-length genome of hcv genotype 1a that elicits cd8⁺ t-cell responses in mice. with the aim to improve the immune response of mva-hcv and because of the importance of interferon (ifn) in hcv infection, we deleted in mva-hcv the vaccinia vir ...201830096846
immunogenicity of propagation-restricted vesicular stomatitis virus encoding ebola virus glycoprotein in guinea pigs.vesicular stomatitis virus (vsv) expressing the ebola virus (ebov) glycoprotein (gp) in place of the vsv glycoprotein g (vsv/ebov-gp) is a promising ebov vaccine candidate which has already entered clinical phase 3 studies. although this chimeric virus was tolerated overall by volunteers, it still caused viremia and adverse effects such as fever and arthritis, suggesting that it might not be sufficiently attenuated. in this study, the vsv/ebov-gp vector was further modified in order to achieve a ...201829869979
whole genome characterization of orthopoxvirus (opv) abatino, a zoonotic virus representing a putative novel clade of old world orthopoxviruses.orthopoxviruses (opvs) are diffused over the complete eurasian continent, but previously described strains are mostly from northern europe, and few infections have been reported from italy. here we present the extended genomic characterization of opv abatino, a novel opv isolated in italy from an infected tonkean macaque, with zoonotic potential. phylogenetic analysis based on 102 conserved opv genes (core gene set) showed that opv abatino is most closely related to the ectromelia virus species ...201830301229
successful establishment of a reverse genetic system for qx-type infectious bronchitis virus and technical improvement of the rescue procedure.in this study, a pathogenic avian infectious bronchitis virus (ibv) qx-type strain yn was successfully rescued by vaccinia virus based reverse genetic technology. ten fragments contiguously spanning the complete ibv genome were amplified and cloned into the vaccinia virus genome by homologous recombination. the full-length genomic cdna was transcribed in vitro, and its transcript was transfected into bhk-21/n cells that could stably express ibv n protein. at 48 h post transfection, the culture m ...201931430502
spi-1 is a missing host-range factor required for replication of the attenuated modified vaccinia ankara (mva) vaccine vector in human cells.modified vaccinia virus ankara (mva) is the leading poxvirus vector for development of vaccines against diverse infectious diseases. this distinction is based on high expression of proteins and good immunogenicity despite an inability to assemble infectious progeny in human cells, which together promote efficacy and safety. nevertheless, the basis for the host-range restriction is unknown despite past systematic attempts to identify the relevant missing viral gene(s). the search for host-range f ...201931145755
chicken embryonic-stem cells are permissive to poxvirus recombinant vaccine vectors.the discovery of mammalian pluripotent embryonic stem cells (esc) has revolutionised cell research and regenerative medicine. more recently discovered chicken esc (cesc), though less intensively studied, are increasingly popular as vaccine substrates due to a dearth of avian cell lines. information on the comparative performance of cesc with common vaccine viruses is limited. using rna-sequencing, we compared cesc transcriptional programmes elicited by stimulation with chicken type i interferon ...201930897824
mva-vectored pentameric complex (pc) and gb vaccines improve pregnancy outcome after guinea pig cmv challenge, but only gb vaccine reduces vertical transmission.(1) background: a congenital cytomegalovirus (ccmv) vaccine is a major research priority, but the essential glycoprotein target(s) remain unclear. we compared cmv gb (gpgb), gh/gl (gp75/gl), and pentameric complex (gppc, composed of gh/gl/gp129/gp131/gp133) vaccines in a guinea pig cmv (gpcmv) congenital infection model. (2) methods: modified vaccinia virus ankara (mva) vaccines expressing gpcmv glycoproteins were used to immunize gpcmv-seronegative, female hartley guinea pigs (three-dose series ...201931739399
recombinant modified vaccinia virus ankara (mva) vaccines efficiently protect cockatiels against parrot bornavirus infection and proventricular dilatation disease.parrot bornaviruses (pabvs) are the causative agents of proventricular dilatation disease (pdd), a chronic and often fatal neurologic disorder in psittaciformes. the disease is widely distributed in private parrot collections and threatens breeding populations of endangered species. thus, immunoprophylaxis strategies are urgently needed. in previous studies we demonstrated a prime-boost vaccination regime using modified vaccinia virus ankara (mva) and newcastle disease virus (ndv) constructs exp ...201931817690
selection of vaccinia virus-neutralizing antibody from a phage-display human-antibody library.although smallpox was eradicated in 1980, it is still considered a potential agent of biowarfare and bioterrorism. smallpox has the potential for high mortality rates along with a major public health impact, eventually causing public panic and social disruption. passive administration of neutralizing monoclonal antibodies (mabs) is an effective intervention for various adverse reactions caused by vaccination and the unpredictable nature of emerging and bioterrorist-related infections. currently, ...201930856707
therapeutic monoclonal antibodies for ebola virus infection derived from vaccinated humans.we describe therapeutic monoclonal antibodies isolated from human volunteers vaccinated with recombinant adenovirus expressing ebola virus glycoprotein (ebov gp) and boosted with modified vaccinia virus ankara. among 82 antibodies isolated from peripheral blood b cells, almost half neutralized gp pseudotyped influenza virus. the antibody response was diverse in gene usage and epitope recognition. although close to germline in sequence, neutralizing antibodies with binding affinities in the nano- ...201930943399
identification of cp77 as the third orthopoxvirus samd9 and samd9l inhibitor with unique specificity for a rodent samd9l.orthopoxviruses (opxvs) have a broad host range in mammalian cells, but chinese hamster ovary (cho) cells are nonpermissive for vaccinia virus (vacv). here, we revealed a species-specific difference in host restriction factor samd9l as the cause for the restriction and identified orthopoxvirus cp77 as a unique inhibitor capable of antagonizing chinese hamster samd9l (chsamd9l). two known vacv inhibitors of samd9 and samd9l (samd9&l), k1 and c7, can bind human and mouse samd9&l, but neither can b ...201930918078
nk cell immune responses differ after prime and boost vaccination.a better understanding of innate responses induced by vaccination is critical for designing optimal vaccines. here, we studied the diversity and dynamics of the nk cell compartment after prime-boost immunization with the modified vaccinia virus ankara using cynomolgus macaques as a model. mass cytometry was used to deeply characterize blood nk cells. the nk cell subphenotype composition was modified by the prime. certain phenotypic changes induced by the prime were maintained over time and, as a ...201930794328
common vole (microtus arvalis) and bank vole (myodes glareolus) derived permanent cell lines differ in their susceptibility and replication kinetics of animal and zoonotic viruses.pathogenesis and reservoir host adaptation of animal and zoonotic viruses are poorly understood due to missing adequate cell culture and animal models. the bank vole (myodes glareolus) and common vole (microtus arvalis) serve as hosts for a variety of zoonotic pathogens. for a better understanding of virus association to a putative animal host, we generated two novel cell lines from bank voles of different evolutionary lineages and two common vole cell lines and assayed their susceptibility, rep ...201931513859
activation of rnase l in egyptian rousette bat-derived roni/7 cells is dependent primarily on oas3 and independent of mavs signaling.bats are reservoirs for many rna viruses that are highly pathogenic in humans yet relatively apathogenic in the natural host. it has been suggested that differences in innate immunity are responsible. the antiviral oas-rnase l pathway is well characterized in humans, but there is little known about its activation and antiviral activity in bats. during infection, oass, upon sensing double-stranded rna (dsrna), produce 2'-5' oligoadenylates (2-5a), leading to activation of rnase l which degrades v ...201931719180
human immunodeficiency virus c.1086 envelope gp140 protein boosts following dna/modified vaccinia virus ankara vaccination fail to enhance heterologous anti-v1v2 antibody response and protection against clade c simian-human immunodeficiency virus challenge.the rv144 human immunodeficiency virus type 1 (hiv-1) vaccine trial showed a strong association between anti-gp70 v1v2 scaffold (v1v2) and anti-v2 hot spot peptide (v2 hs) antibody responses and reduced risk of hiv infection. accordingly, a primary goal for hiv vaccines is to enhance the magnitude and breadth of v1v2 and v2 hs antibody responses in addition to neutralizing antibodies. here, we tested the immunogenicity and efficacy of hiv-1 c.1086 gp140 boosts administered sequentially after pri ...201931341049
bridging vaccine-induced hiv-1 neutralizing and effector antibody responses in rabbit and rhesus macaque animal models.studies in animal models are essential prerequisites for clinical trials of candidate hiv vaccines. small animals, such as rabbits, are used to evaluate promising strategies prior to further immunogenicity and efficacy testing in nonhuman primates. our goal was to determine how hiv-specific vaccine-elicited antibody responses, epitope specificity, and fc-mediated functions in the rabbit model can predict those in the rhesus macaque (rm) model. detailed comparisons of the hiv-1-specific igg respo ...201930842326
clade c hiv-1 envelope vaccination regimens differ in their ability to elicit antibodies with moderate neutralization breadth against genetically diverse tier 2 hiv-1 envelope variants.the goals of preclinical hiv vaccine studies in nonhuman primates are to develop and test different approaches for their ability to generate protective immunity. here, we compared the impact of 7 different vaccine modalities, all expressing the hiv-1 1086.c clade c envelope (env), on (i) the magnitude and durability of antigen-specific serum antibody responses and (ii) autologous and heterologous neutralizing antibody capacity. these vaccination regimens included immunization with different comb ...201930651354
novel modified vaccinia virus ankara vector expressing anti-apoptotic gene b13r delays apoptosis and enhances humoral responses.modified vaccinia virus ankara (mva), an attenuated poxvirus, has been developed as a potential vaccine vector for use against cancer and multiple infectious diseases, including human immunodeficiency virus (hiv). mva is highly immunogenic and elicits strong cellular and humoral responses in preclinical models and humans. however, there is potential to further enhance the immunogenicity of mva, as mva-infected cells undergo rapid apoptosis, leading to faster clearance of recombinant antigens and ...201930541829
priming with a potent hiv-1 dna vaccine frames the quality of immune responses prior to a poxvirus and protein boost.the use of heterologous immunization regimens and improved vector systems has led to increases in immunogenicity of hiv-1 vaccine candidates in nonhuman primates. in order to resolve interrelations between different delivery modalities, three different poxvirus boost regimens were compared. three groups of rhesus macaques were each primed with the same dna vaccine encoding gag, pol, nef, and gp140. the groups were then boosted with either the vaccinia virus strain nyvac or a variant with improve ...201930429343
replication-competent nyvac-kc yields improved immunogenicity to hiv-1 antigens in rhesus macaques compared to nonreplicating nyvac.as part of the continuing effort to develop an effective hiv vaccine, we generated a poxviral vaccine vector (previously described) designed to improve on the results of the rv144 phase iii clinical trial. the construct, nyvac-kc, is a replication-competent, attenuated recombinant of the vaccinia virus strain nyvac. nyvac is a vector that has been used in many previous clinical studies but is replication deficient. here, we report a side-by-side comparison of replication-restricted nyvac and rep ...201930429340
cd8 t cells show protection against highly pathogenic simian immunodeficiency virus (siv) after vaccination with siv gene-expressing bcg prime and vaccinia virus/sendai virus vector boosts.toward development of a dual vaccine for human immunodeficiency virus (hiv)-1 and tuberculosis infections, we developed a urease-deficient bacillus calmette-guérin (bcg) strain tokyo172 (bcgδurease) to enhance its immunogenicity. bcgδurease expressing simian immunodeficiency virus (siv) gag induced bcg antigen-specific cd4+ and cd8+ t cells more efficiently, and more gag-specific cd8+ t cells. we evaluated its protective efficacy against siv infection in cynomolgus monkeys of asian origin, shown ...202033087465
innate and secondary humoral responses are improved by increasing the time between mva vaccine immunizations.comprehending the mechanisms behind the impact of vaccine regimens on immunity is critical for improving vaccines. indeed, the time-interval between immunizations may influence b and t cells, as well as innate responses. we compared two vaccine schedules using cynomolgus macaques immunized with an attenuated vaccinia virus. two subcutaneous injections 2 weeks apart led to an impaired secondary antibody response and similar innate myeloid responses to both immunizations. in contrast, a delayed bo ...202032218996
the vaccinia virus based sementis copenhagen vector vaccine against zika and chikungunya is immunogenic in non-human primates.the sementis copenhagen vector (scv) is a new vaccinia virus-derived, multiplication-defective, vaccine technology assessed herein in non-human primates. indian rhesus macaques (macaca mulatta) were vaccinated with a multi-pathogen recombinant scv vaccine encoding the structural polyproteins of both zika virus (zikv) and chikungunya virus (chikv). after one vaccination, neutralising antibody responses to zikv and four strains of chikv, representative of distinct viral genotypes, were generated. ...202032550013
evaluation of heterologous prime-boost vaccination strategies using chimpanzee adenovirus and modified vaccinia virus for tb subunit vaccination in rhesus macaques.tuberculosis (tb) still is the principal cause of death from infectious disease and improved vaccination strategies are required to reduce the disease burden and break tb transmission. here, we investigated different routes of administration of vectored subunit vaccines based on chimpanzee-derived adenovirus serotype-3 (chad3) for homologous prime-boosting and modified vaccinia virus ankara (mva) for heterologous boosting with both vaccine vectors expressing the same antigens from mycobacterium ...202032435513
a new oncolytic vacciniavirus augments antitumor immune responses to prevent tumor recurrence and metastasis after surgery.local recurrence and remote metastasis are major challenges to overcome in order to improve the survival of patients with cancer after surgery. oncolytic viruses are a particularly attractive option for prevention of postsurgical disease as they offer a non-toxic treatment option that can directly target residual tumor deposits and beneficially modulate the systemic immune environment that is suppressed post surgery and allows residual disease escape from control. here, we report that a novel va ...202032217766
molecular and genomic characterization of a novel equine molluscum contagiosum-like virus.cases of pox-like lesions in horses and donkeys have been associated with poxviruses belonging to different genera of the family poxviridae. these include the orthopoxviruses vaccinia virus (vacv), horsepoxvirus (hpxv) and cowpoxvirus (cpxv), as well as a potentially novel parapoxvirus and molluscum contagiosum virus (mocv). however, with the exception of vacv, hpxv and cpxv, the genomic characterization of the causative agents remains largely elusive with only single short genome fragments avai ...202031922947
molecular and biological characterization of a cervidpoxvirus isolated from moose with necrotizing dermatitis.cervidpoxvirus is one of the more recently designated genera within the subfamily chordopoxvirinae, with deerpox virus (dpv) as the only recognized species to date. in this study, the authors describe spontaneous disease and infection in the north american moose (alces americanus) by a novel cervidpoxvirus, here named moosepox virus (mpv). three 4-month-old moose calves developed a multifocal subacute-to-chronic, necrotizing, suppurative-to-granulomatous dermatitis that affected the face and the ...202032096438
repair of a previously uncharacterized second host-range gene contributes to full replication of modified vaccinia virus ankara (mva) in human cells.modified vaccinia virus ankara (mva), a widely used vaccine vector for expression of genes of unrelated pathogens, is safe, immunogenic, and can incorporate large amounts of added dna. mva was derived by extensively passaging the chorioallantois vaccinia virus ankara (cva) vaccine strain in chicken embryo fibroblasts during which numerous mutations and deletions occurred with loss of replicative ability in most mammalian cells. restoration of the deleted c12l gene, encoding serine protease inhib ...202032019881
exposure of free-ranging capybaras (hydrochoerus hydrochaeris) to the vaccinia virus.the aim of this study was to evaluate the possibility of free-ranging animals/hunting dogs as sources of infection in the vaccinia virus (vacv) transmission chain. serological, cell culture and molecular assays were conducted in 56 free-ranging animals and 22 hunting dogs. elisa/neutralizing assays showed that two (2.5%) capybaras (hydrochoerus hydrochaeris) had anti-opv positive antibodies, while all samples tested negative through pcr/cell culture. after being hit by cars on roads, capybaras t ...202031574216
field trial vaccination against cowpox in two alpaca herds.in europe, cowpox virus (cpxv) infection in south american camelids occurs as a so-called spill-over infection. although infected animals generally have a mild form of the disease and survive, cases of fatal generalised cpxv infection have also been described. prevention by prophylactic vaccination is the only way to protect animals from disease. in the present study, modified vaccinia virus ankara (mva) vaccine, which has been successfully used in many animal species, was used in a prime-boost ...202032093320
sars_cov2 rbd gene transcription cannot be driven by cmv promoter.cytomegalovirus (cmv) promoter drives various gene expression and yields sufficient protein for further functional investigation. receptor binding domain (rbd) on spike protein of the sars_cov2 is the most critical portal for virus infection. thus native conformational rbd protein may facilitate biochemical identification of rbd and provide valuable support of drug and vaccine design for curing covid-19. we attempted to express rbd under cmv promoter in vitro, but failed. rbd-specific mrna canno ...202133711560
educational approach to prevent the burden of vaccinia virus infections in a bovine vaccinia endemic area in brazil.bovine vaccinia (bv), caused by vaccinia virus (vacv), is a zoonotic disease characterized by exanthematous lesions on the teats of dairy cows and the hands of milkers, and is an important public health issue in brazil and south america. bv also results in economic losses to the dairy industry, being a burden to the regions involved in milk production. in the past 20 years, much effort has been made to increase the knowledge regarding bv epidemiology, etiologic agents, and interactions with the ...202133922509
genomic diversity of vaccinia virus strain cantagalo isolated in southeastern brazil during the early years of the outbreak, 1999-2006.outbreaks of a vesiculopustular disease in dairy cattle and milkers have been frequently reported in brazil since 1999 when the vaccinia virus strain cantagalo was first isolated in the state of rio de janeiro. however, the genomic diversity of the viral isolates associated with these outbreaks is not well known, particularly in the southeastern states that represent the focal point of virus spread to other regions. here, we report the genomic sequences and an analysis of the polymorphic site pr ...202133566940
time-course transcriptome analysis of host cell response to poxvirus infection using a dual long-read sequencing approach.in this study, we applied two long-read sequencing (lrs) approaches, including single-molecule real-time and nanopore-based sequencing methods to investigate the time-lapse transcriptome patterns of host gene expression as a response to vaccinia virus infection. transcriptomes determined using short-read sequencing approaches are incomplete because these platforms are inefficient or fail to distinguish between polycistronic rnas, transcript isoforms, transcriptional start sites, as well as trans ...202134167576
development and characterization of a highly sensitive nanoluciferase-based immunoprecipitation system for the detection of anti-influenza virus ha antibodies.antibody detection is crucial for monitoring host immune responses to specific pathogen antigens (ags) and evaluating vaccine efficacies. the luciferase immunoprecipitation system (lips) was developed for sensitive detection of ag-specific antibodies in sera from various species. in this study, we describe nanolips, an improved lips assay based on nanoluciferase (nluc), and employ the assay for monitoring antibody responses following influenza virus infection or vaccination. we generated recombi ...202133980684
a systemically deliverable vaccinia virus with increased capacity for intertumoral and intratumoral spread effectively treats pancreatic cancer.pancreatic cancer remains one of the most lethal cancers and is refractory to immunotherapeutic interventions. oncolytic viruses are a promising new treatment option, but current platforms demonstrate limited efficacy, especially for inaccessible and metastatic cancers that require systemically deliverable therapies. we recently described an oncolytic vaccinia virus (vv), vvlδtkδn1l, which has potent antitumor activity, and a regime to enhance intravenous delivery of vv by pharmacological inhibi ...202133500259
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