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activity of a fowlpox virus late gene promoter in vaccinia and fowlpox virus recombinants.characterization of a late promoter of fowlpox virus (fpv) and a study of its activity in fpv and vaccinia virus (vv) was carried out. the 5'-mrna start site of the fpv late gene mapped to a taaat sequence near the translation start site (atg). a cloned dna fragment of fpv genome (pfl1) comprising of the 5'-end of the late gene was used to express the lacz gene of e. coli in fpv and vv recombinants. a comparative analysis of beta-galactosidase (bg) expression from the lacz gene under the control ...19902165765
production and characterisation of a monoclonal antibody to human papillomavirus type 16 using recombinant vaccinia virus.a monoclonal antibody was raised against the major capsid protein l1 of human papillomavirus type 16, using a recombinant vaccinia virus that expresses the l1 protein, as a target for screening. this antibody, designated camvir-1, reacted with a 56 kilodalton protein in cells infected with l1-vaccinia virus, and the protein was present in a predominantly nuclear location. the antibody also detects the hpv-16 l1 antigen in formalin fixed, paraffin wax embedded biopsy specimens and on routine cerv ...19902166093
purification and use of vaccinia virus messenger rna capping enzyme. 19902166212
guidelines of bundesgesundheitsamt (bga; german federal health office) and deutsche vereinigung zur bekämpfung der viruskrankheiten e.v. (dvv; german association for the control of virus diseases) for testing the effectiveness of chemical disinfectants against viruses. 19902166529
inhibition of antigen-induced and interleukin-2-induced proliferation of bovine peripheral blood leukocytes by inactivated bovine herpes virus 1.the mechanism by which bovine herpesvirus 1 (bhv-1) predisposes cattle to bacterial pneumonia was investigated by using an in vitro system to demonstrate immunosuppression. at a multiplicity of infection of 0.001, live or inactivated bhv-1 induced a 50% inhibition of the proliferative response of peripheral blood mononuclear leukocytes to antigen (vaccinia virus in vaccinia virus-immunized cattle which were bhv-1 negative) or interleukin-2. at this same multiplicity of infection, the mitogen-ind ...19902166810
the glycoprotein products of varicella-zoster virus gene 14 and their defective accumulation in a vaccine strain (oka).many characteristics of the putative protein encoded by varicella-zoster virus (vzv) open reading fram (orf) 14 indicate that it is a glycoprotein, which has been designated gpv. to identify the protein products of the gene, the coding sequences were placed under the control of the vaccinia virus p7.5 promoter and recombinant vaccinia viruses were constructed. heterogeneous polypeptides with molecular weights of 95,000 to 105,000 (95k to 105k polypeptides) were expressed in cells infected by a v ...19902166829
induction of neutralizing antibodies against bovine leukosis virus in rabbits by vaccination with recombinant vaccinia virus expressing bovine leukosis virus envelope glycoprotein.three kinds of recombinant vaccinia virus (rvv)--mo-ha/ati, lo1-ha/ati and mo-ha/7.5kd--expressing bovine leukosis virus (blv) envelope glycoprotein (gp60) were constructed. the blv envelope gene of rvv mo-ha/ati and lo1-ha/ati or of rvv mo-ha/7.5kd was expressed under control of the promoter of a-type inclusion body (ati) protein gene of cow-pox virus or vaccinia virus 7.5-kd protein gene, respectively. the vaccinia virus strain, lc16mo, was used as vector for rvv mo-ha/ati and mo-ha/7.5kd, and ...19902167028
ocular safety and efficacy of an hsv-1 gd vaccine during primary and latent infection.one potential complication of systemic herpes simplex virus (hsv) vaccination is that subsequent ocular infection may lead to increased immunogenic corneal scarring. therefore, v52, a genetically engineered vaccinia virus that expresses the hsv-1 glycoprotein gd, was tested for ocular safety and for protection against ocular challenge with a stromal-disease-producing strain (mckrae) of hsv-1. to maximize immune response, rabbits were vaccinated by a series of inoculations. v52-vaccinated rabbits ...19902167298
synthesis of functional mrna in mammalian cells by bacteriophage t3 rna polymerase.we found that the 5' nontranslated leader sequence from encephalomyocarditis virus (emcv) allowed transcripts that were synthesized by the t3 rna polymerase in mammalian cells to be translated in a cap-independent fashion. stable mouse cell lines that carry the t3 rna polymerase gene expressed the chloramphenicol acetyltransferase (cat) gene under the control of a phage promoter when the cat gene was fused to the emcv leader and introduced into the cells by transient dna uptake. the level of gen ...19902167433
[verification of the safety, inoculability, reactogenicity and antigenic properties of a live recombinant smallpox-hepatitis b vaccine in an experiment in volunteers].trials of the first soviet live recombinant smallpox-hepatitis b vaccine (shbv) in volunteers (20 men aged 18-20 years) showed its safety, good "take"-rate, and lower reactogenicity as compared with the standard smallpox vaccine (livp strain). smallpox virus-neutralizing antibodies in response to shbv were produced as well as in response to the smallpox vaccine. revaccination of human subjects with smallpox vaccine and shbv 45 days after the previous vaccination resulted in antibody booster to v ...19902167564
the immunogenicity of vp7, a rotavirus antigen resident in the endoplasmic reticulum, is enhanced by cell surface expression.the glycoprotein vp7, the major serotype antigen of rotaviruses, is localized to the endoplasmic reticulum (er) of the cell, where it is retained as a membrane-associated protein before assembly into mature virus particles. wild-type vp7 expressed by a recombinant vaccinia virus was also located internally and was poorly antigenic. using recombinant techniques, a correctly processed, secreted form of vp7 (s.c. stirzaker and g.w. both, cell 56:741-747, 1989) was modified by addition to its c term ...19902168965
the e1 glycoprotein of an avian coronavirus is targeted to the cis golgi complex.it was previously reported that the e1 protein of an avian coronavirus was targeted to the juxtanuclear region in cos cells expressing the protein from cloned cdna, suggesting that the protein contains information for targeting to the golgi complex. the first of three membrane-spanning domains was required for intracellular targeting, because a mutant e1 (delta m1,2) lacking this domain was delivered to the plasma membrane. we have used immunoelectron microscopy to localize the wild-type e1 prot ...19902169615
specific dna cleavage and binding by vaccinia virus dna topoisomerase i.cleavage of a defined linear duplex dna by vaccinia virus dna topoisomerase i was found to occur nonrandomly and infrequently. approximately 12 sites of strand scission were detected within the 5372 nucleotides of puc19 dna. these sites could be classified as having higher or lower affinity for topoisomerase based on the following criteria. higher affinity sites were cleaved at low enzyme concentration, were less sensitive to competition, and were most refractory to religation promoted by salt, ...19902170398
apolipoprotein a-i and its amphipathic helix peptide analogues inhibit human immunodeficiency virus-induced syncytium formation.the envelope (membrane) glycoprotein of hiv is essential for virus attachment and entry into host cells. additionally, when expressed on the plasma membrane of infected cells, the envelope protein is responsible for mediating cell-cell fusion which leads to the formation of multinucleated giant cells, one of the major cytopathic effects of hiv infections. the envelope glycoproteins of hiv contain regions that can fold into amphipathic alpha-helixes, and these regions have been suggested to play ...19902170446
increased antibody responses to human papillomavirus type 16 l1 protein expressed by recombinant vaccinia virus lacking serine protease inhibitor genes.the l1 gene of human papillomavirus type 16 (hpv-16) driven by the vaccinia virus major late 4b gene promoter has been inserted into three different sites of the vaccinia virus genome. insertion into the thymidine kinase (tk) gene was achieved by selection of tk- mutants in budr on tk- cells. insertion into two vaccinia virus serine protease inhibitor (serpin) genes was achieved by co-insertion of the escherichia coli xanthine guanine phosphoribosyltransferase gene linked to the vaccinia virus 7 ...19902170578
wallace p. rowe lecture. poxviruses of laboratory animals.currently the subfamily chordopoxvirinae, the poxviruses of vertebrates, is subdivided into eight genera, containing some 20-30 species; an inexact figure because the birdpox viruses have not yet been properly investigated taxonomically. i have discussed seven species belonging to three genera, all of which have caused infection and usually disease in mammals commonly used in the laboratory. the list could have been extended had i included chickens and swine as laboratory animals, for that would ...19902170745
complementation of a vesicular stomatitis virus glycoprotein g mutant with wild-type protein expressed from either a bovine papilloma virus or a vaccinia virus vector system.using a complementation assay, we have evaluated the potential of two eukaryotic expression systems to produce functional virus proteins. the first expression system was based on a bovine papilloma virus (bpv) eukaryotic expression vector which contained a copy of the gene for the membrane glycoprotein g of vesicular stomatitis virus (vsv). this vector was transfected into a mouse cell line, and transformed cell clones constitutively expressing vsv g protein were selected. these cell clones were ...19902171187
vaccinia virus host range genes.a gene encoding an 18-kda polypeptide (orf c7l) located in the vaccinia virus hindiii c fragment was shown to be functionally equivalent to previously described host range gene (orf k1l) spanning the hindiii k/m fragment junction. either c7l or k1l host range gene is necessary and sufficient by itself to allow replication of vaccinia virus on human cells. deletion of both c7l and k1l genes from the wild-type vaccinia genome is required to derive a virus deficient for replication on human cells. ...19902171207
murine cytotoxic t lymphocytes specific for herpes simplex virus type 1 recognize the immediate early protein icp4 but not icp0.vaccinia virus recombinants expressing the herpes simplex virus type 1 (hsv-1) genes encoding icp0 or icp4 were used to identify the precise target antigen(s) of murine anti-viral cytotoxic t lymphocytes (ctl) specific for the non-structural immediate early proteins. these studies revealed that hsv-1-specific ctl, restricted to class i major histocompatibility complex genes of the h-2k haplotype but not the h-2d or h-2b haplotypes, would lyse autologous cells expressing icp4. hsv-1-specific ctl ...19902172452
a complex between the mhc class i homologue encoded by human cytomegalovirus and beta 2 microglobulin.human cytomegalovirus (hcmv) is a ubiquitous pathogen that persists in the host and can cause severe disease in the immunocompromised individual or in the fetus. analysis of the nucleotide sequence of the virus genome has revealed the presence of an open reading frame whose predicted translation product has homology with the heavy chain of the major histocompatibility complex (mhc) class i molecule of higher eukaryotes, and the observed sequence homology was given additional significance by the ...19902172831
[effects of estrogen treatment on the antiviral properties of murine kupffer cells].adult female mice were administered 17 beta-oestradiol at pharmacological dosages by subcutaneous injections. histopathological examination revealed an increase in cells of the mononuclear lineage infiltrating the sinusoids of the liver. in vitro culture of kupffer cells demonstrated that the treatment did not alter their antiviral properties but did rather induce an activation of their non-specific immune properties. this activation might be beneficial or deleterious for the infected host and m ...19902172894
expression and secretion of pres containing hepatitis b surface antigen in vaccinia virus system.the expression and secretion of pres containing hepatitis b surface antigen in vaccinia virus system was investigated. the human tk- 143 cells were infected with the recombinant vaccinia viruses vtms-1 or vtls-1. cells infected with vtms-1, which contains the pres2 + s gene, produced pres2 containing middle hbsag proteins. similarly, cells produced pres1 containing large hbsag proteins upon infection with vtls-1, which carries the pres1 + pres2 + s gene. the expression products could be secreted ...19902173617
proteins encoded by the bovine papillomavirus e1 open reading frame: expression in heterologous systems and in virally transformed cells.the e1 open reading frame (orf) of bovine papillomavirus type 1 is required for the persistence of viral genomes as multicopy plasmid molecules in transformed rodent fibroblasts. e1 has been reported to contain two separate complementation groups (m and r, corresponding to n- and c-terminal domains, respectively) which regulate viral replication. however, e1 behaves as a single gene with respect to cell transformation and viral transcription. we examined the proteins translated from the entire o ...19902173778
recombinant fowlpox viruses inducing protective immunity against newcastle disease and fowlpox viruses.the haemagglutinin-neuraminidase gene of newcastle disease virus was inserted into a non-essential region of the fowlpox virus genome and expressed under control of the vaccinia virus 7.5 kda polypeptide gene promoter. immunization with the recombinant fowlpox virus elicited protective immunity in chickens against both virulent newcastle disease and fowlpox virus infection.19902174598
role of coexpression of il-2 and herpes simplex virus proteins in recombinant vaccinia virus vectors on levels of induced immunity.vaccinia virus recombinants co-expressing the genes for il-2 and hsv-1 gc or gd were used to study the potential adjuvant effects of il-2 on immunity to hsv-1. in addition, constructs were produced which, while expressing il-2, encoded for reduced levels of hsv-1 gc. studies with mice revealed that gc and gd could stimulate hsv-1 immunity as measured by neutralizing antibody (na), lymphoproliferation and viral clearance. il-2 enhancement of na and lymphoproliferation was observed only in those g ...19902175194
nucleotide sequence comparison of homologous genomic regions from variola, monkeypox, and vaccinia viruses.the nucleotide sequences of homologous regions from the genomes of variola major strain harvey, variola minor strains butler and garcia, and monkeypox strain denmark were determined. the nucleotide sequences were compared to the homologous region in vaccinia virus which contains part of one of the genes involved in determining host range. two major differences were detected; these corresponded to a deletion in the promoter region and the presence of a premature stop codon. it was concluded that ...19902176671
analysis of the fowlpox virus genome region corresponding to the vaccinia virus d6 to a1 region: location of, and variation in, non-essential genes in poxviruses.the dna sequence of the fowlpox virus genome corresponding to the vaccinia virus d6 to a1 region has been determined. translation of this sequence reveals fowlpoxvirus gene homologues corresponding to the d6, d7, d9, d10, d11, d12, d13 and a1 genes of vaccinia virus. in contrast, no gene homologue for the non-essential vaccinia virus d8 gene was present in fowlpox virus. instead, a gene transcribed from the opposite strand to the vaccinia virus d8 gene showing no homology to any previously seque ...19902177083
evaluation of thymidine kinase and neomycin phosphotransferase ii positive selection systems for recovery of genetically atypical and recombinant dna vaccine viruses.dominant phenotypic marker cell culture selection systems were evaluated for the recovery of thymidine kinase positive (tk+), thymidine kinase negative (tk-), and neomycin phosphotransferase ii positive (npt ii+) viruses. from 1 to 100 pfu of each marker-positive virus was diluted into 10(6) pfu of marker-negative background virus prior to selection. all three selection systems recovered 100 or fewer pfu of marked virus from the background population (10(-4) fractional level). in some instances, ...19902178350
mutagenic activation of 2-amino-3-methylimidazo[4,5-f]quinoline by complementary dna-expressed human liver p-450.eight forms of human liver microsomal p-450 were individually expressed in human hepatoma hep g2 cells with a vaccinia virus cdna expression system. using the ames test, each expressed p-450 was examined for its ability to activate to mutagenic products the compounds, 2-amino-3-methylimidazo[4,5-f]quinoline, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, and 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline, respectively. three forms of human p-450 sig ...19902180561
removal of cryptic poxvirus transcription termination signals from the human immunodeficiency virus type 1 envelope gene enhances expression and immunogenicity of a recombinant vaccinia virus.the in vivo role of the proposed poxvirus early transcription termination signal tttttnt was confirmed by analysis of the rna species made by recombinant vaccinia viruses. premature transcription termination occurred following each of two tttttnt sequences present naturally within the coding region of the human immunodeficiency virus type 1 envelope gene. alteration of the tttttnt sequences, without changing the encoded amino acids, resulted in production of full-length early mrnas, improved pro ...19902182912
hemadsorption and fusion inhibition activities of hemagglutinin analyzed by vaccinia virus mutants.vaccinia virus ihd-j strain induces hemagglutinin (ha) on the surface membrane of infected cells and does not elicit cell-cell fusion (f-). we isolated 21 independent hemadsorption-negative (had-) mutant viruses from ihd-j and five had+ revertants from one of these mutants. of the 21 mutants, 19 that synthesized either no or little ha at the cell surface caused cell-cell fusion (f+), whereas none of the five revertants that synthesized ha at the cell surface induced cell-cell fusion. furthermore ...19902183466
human immunodeficiency virus-like, nonreplicating, gag-env particles assemble in a recombinant vaccinia virus expression system.we report the assembly of human immunodeficiency virus (hiv)-like particles in african green monkey kidney cells coinfected with two recombinant vaccinia viruses, one carrying the hiv-1 gag and protease genes and the other the env gene. biochemical analysis of particles sedimented from culture supernatants of doubly infected cells revealed that they were composed of gag proteins, primarily p24, as well as the env proteins gp120 and gp41. thin-section immunoelectron microscopy showed that these p ...19902186175
variable proteolytic cleavage of gag precursor expressed after infection of several cell lines with an hiv-2 gag-pol recombinant vaccinia virus. 19902187498
vaccinia virus vectors: new strategies for producing recombinant vaccines.the development and continued refinement of techniques for the efficient insertion and expression of heterologous dna sequences from within the genomic context of infectious vaccinia virus recombinants are among the most promising current approaches towards effective immunoprophylaxis against a variety of protozoan, viral, and bacterial human pathogens. because of its medical relevance, this area is the subject of intense research interest and has evolved rapidly during the past several years. t ...19902187593
recognition of human t cell leukemia virus type i (htlv-i) gag and px gene products by mhc-restricted cytotoxic t lymphocytes induced in rats against syngeneic htlv-i-infected cells.we established rat t cell lines expressing human t cell leukemia virus type i (htlv-i) ag from inbred strains of rats, wka/h, da, and f344, to study ctl response against the htlv-i-infected cells. htlv-i-specific ag expressed in these rat cells were htlv-i gag ag, p19, p24, and p15, and px ag, p40tax and p27rex, but not env ag, as determined by immunofluorescence and immunoblot assays. by immunization of rats with syngeneic htlv-i-infected cells, ctl against syngeneic htlv-i-infected cells and a ...19902187927
protein synthesis initiation factor modifications during viral infections: implications for translational control.infection of tissue culture cells with certain viruses results in the shutoff of host cell protein synthesis. we have examined virally infected cell lysates using two-dimensional gel electrophoresis and immunoblotting to ascertain whether initiation factor protein modifications are correlated with translational repression. moderate increases in eukaryotic initiation factor (eif)-2 alpha phosphorylation are detected in reovirus- and adenovirus-infected cells, as reported previously (samuel et al. ...19902188834
intracellular processing and immunogenicity of the envelope proteins of human t-cell leukemia virus type i that are expressed from recombinant vaccinia viruses.two types of recombinant vaccinia viruses (vvs) expressing the env gene of the human t-cell leukemia virus type i (htlv-i) were reported previously. one recombinant vv, wr-proenv1, synthesized the authentic env protein. in the other recombinant vv, wr-env17, the env gene was inserted within the signal sequence of the vv hemagglutinin (ha) gene, so that the reading frame for the env gene was in phase with that for the ha gene. comparative studies were performed on the mode of expression and proce ...19902189258
early transcription factor subunits are encoded by vaccinia virus late genes.the vaccinia virus early transcription factor (vetf) was shown to be a virus-encoded heterodimer. the gene for the 82-kda subunit was identified as open reading frame (orf) a8l, based on the n-terminal sequence of factor purified by using dna-affinity magnetic beads. the 70-kda subunit of vetf was refractory to n-terminal analysis, and so n-terminal sequences were obtained for three internal tryptic peptides. all three peptides matched sequences within orf d6r. orfs a8l and d6r are located withi ...19902190222
inducible gene expression from vaccinia virus vectors.a system for inducible gene expression by vaccinia virus (vv) vectors utilizing the escherichia coli lac i repressor/operator system is described. a vv recombinant that expresses the lac i repressor protein from the constitutively active 7.5k promoter was constructed. gel retardation experiments showed that a protein present in extracts of cells infected with this virus, but not wild-type virus, bound to the lac operator and that this binding was inhibited by iptg. a series of vv recombinants we ...19902191497
yeast kex1 protease cleaves a prohormone processing intermediate in mammalian cells.a vaccinia virus vector was used to express the yeast kex1 gene, which encodes a prohormone carboxypeptidase specific for the removal of basic amino acids from prohormone processing intermediates, in mammalian cells. when produced in bsc-40 cells, kex1p was localized to the perinuclear region and conferred a large increase in enzymatic activity characteristic of this carboxypeptidase. expression of the kex1 gene together with the yeast kex2 gene, which encodes a prohormone endopeptidase specific ...19902193026
identification and nucleotide sequence of the gene encoding a surface antigen induced by vaccinia virus.the gene encoding the specific antigen (s antigen) induced on the surface of vaccinia virus-infected cells was mapped on the viral genome by the construction of recombinant viruses. the s gene is located in the 1.5-kbp xbai-accii fragment between 11.1 and 12.6 kbp from the right end of the viral genome. nucleotide sequencing of the fragment revealed an open reading frame (orf) of 351 codons capable of producing a polypeptide with a molecular weight (mw) of 40,701 da. mapping of the s mrna by pri ...19902196742
regulation of vaccinia virus transcription. 19902197987
poxvirus recombinant vaccines.the poxvirus family, subfamily chordopoxviridae, contains six genera. the basic criteria distinguishing membership in a genus is cross-neutralization and host-range predilection. members of the genus orthopoxvirus (vaccinia) and of the genus avipoxvirus (fowlpox) have been genetically engineered to express genes from heterologous pathogens, thus providing a means of assessing these recombinant viruses as live recombinant vaccines. vaccinia virus recombinants which have a broad host range can be ...19902198832
response of monkeys to vaccination with recombinant vaccinia virus which coexpress hiv gp160 and human interleukin-2.immunization of two macaque monkeys with recombinant vaccinia viruses encoding the env gene of hiv-1 (vv-gp160) resulted in demonstrable levels of gp160, gp120 and gp41-specific immunoglobulins in both animals. the virus used to immunize one of the monkeys additionally expressed the human il-2 gene, which encoded human il-2 (vv-gp160-il-2). no toxic side-effects of vaccine-delivered il-2 were observed. despite marked attenuation of virulence by the coexpressed lymphokine, the levels of vector-sp ...19902200748
function of semliki forest virus e3 peptide in virus assembly: replacement of e3 with an artificial signal peptide abolishes spike heterodimerization and surface expression of e1.the semliki forest virus spike glycoproteins e1 and p62 form a heterodimeric complex in the endoplasmic reticulum (er) and are transported as such to the cell surface. in the mature virus particle, the heterodimeric association of e1 and e2 (the cleavage product of p62) is maintained, but as a more labile and acid-sensitive oligomer than the e1-p62 complex. the e3 peptide forms the n-terminal part of the p62 precursor and carries the signal for the translocation of p62 into the lumen of the er. ...19902200886
cytoplasmic expression system based on constitutive synthesis of bacteriophage t7 rna polymerase in mammalian cells.a mouse cell line that constitutively synthesizes the bacteriophage t7 rna polymerase was constructed. fluorescence microscopy indicated that the t7 rna polymerase was present in the cytoplasmic compartment. the system provided, therefore, a unique opportunity to study structural elements of mrna that affect stability and translation. the in vivo activity of the bacteriophage polymerase was demonstrated by transfection of a plasmid containing the chloramphenicol acetyltransferase (cat) gene flan ...19902204064
regulated expression of nuclear genes by t3 rna polymerase and lac repressor, using recombinant vaccinia virus vectors.recombinant vaccinia viruses that express the bacteriophage t3 rna polymerase (vv-t3pol) or the escherichia coli lac repressor (vv-laci) under control of the early-late vaccinia promoter p7.5 were constructed. to determine whether phage polymerase and lac repressor can function in the nucleus of mammalian cells, the bacterial chloramphenicol acetyltransferase (cat) gene was cloned downstream of a t3 promoter (pt3-cat) or downstream of a t3 promoter-lac operator fusion element (pt3olac-cat), and ...19902204724
eukaryotic dna ligases.recent studies on eukaryotic dna ligases are briefly reviewed. the two distinguishable enzymes from mammalian cells, dna ligase i and dna ligase ii, have been purified to homogeneity and characterized biochemically. two distinct dna ligases have also been identified in drosophila melanogaster embryos. the genes encoding dna ligases from schizosaccharomyces pombe, saccharomyces cerevisiae and vaccinia virus have been cloned and sequenced. these 3 proteins exhibit about 30% amino acid sequence ide ...19902204827
the metabolism of 1-nitropyrene by human cytochromes p450.the metabolism of [3h]1-nitropyrene by specific forms of human cytochrome p450 was investigated in vitro using vaccinia virus expression of p450 cdnas in hepg2 cells. cell lysates were injected individually with recombinant vaccinia virus containing human p450 cdna (p450 ia2, iia3, iib7, iic8, iic9, iid6, iie1, iif1, iiia3, iiia4, iiia5 and ivb1). only iiia3 and iiia4 demonstrated significant activity in the c-oxidation of 1-nitropyrene. the principal metabolite from both p450 forms was 1-nitrop ...19902205409
use of vaccinia rabies recombinant for oral vaccination of wildlife.a vaccinia rabies recombinant virus was constructed and shown to induce the synthesis of rabies virus glycoprotein in infected cells and to induce rabies virus neutralizing antibodies and protection in susceptible animals. active when orally administered, this recombinant is a good candidate for the development of vaccines for wild animal rabies vectors. this recombinant was found stable, safe for target and non-target animal species, and protective for most of the rabies vectors. after extensiv ...19902205969
antitumor efficacy of vaccinia virus-modified tumor cell vaccine.the antitumor efficacies of vaccinia virus-modified tumor cell vaccines were examined in murine syngeneic mh134 and x5563 tumor cells. uv-inactivated vaccinia virus was inoculated i.p. into c3h/hen mice that had received whole body x-irradiation at 150 rads. after 3 weeks, the vaccines were administered i.p. 3 times at weekly intervals. one week after the last injection, mice were challenged i.p. with various doses of syngeneic mh134 or x5563 viable tumor cells. four methods were used for prepar ...19902208158
adenosylmethionine decarboxylase inhibitors--lack of activity against cytopathic effects of hiv.many bis(amidinohydrazones) are potent inhibitors of adenosylmethionine decarboxylase (adometdc), a key enzyme of polyamine biosynthesis, and some of them are also known to be powerful antiviral agents. therefore, seven bis(amidinohydrazones), including the two most potent inhibitors of eukaryotic adometdc so far reported, were screened for antiviral activity against the human immunodeficiency virus (hiv). the screening was performed by incubating susceptible human leukemia cells in microculture ...19902209170
western blot analysis of serological responses following immunization with vaccinia viral lysates of melanoma cells.western blot analysis was used to characterize the antibody response of melanoma patients during immunization with vaccinia-virus-induced melanoma cell lysates (vmcl). strong antibody responses were detectable within 4 weeks from commencement of immunization against different fractions (from 3 to 10) in blots of vmcl. the approximate mw of the main fractions identified were 100, 91, 85, 77, 64-66, 49, 46, 43 and 34-36 kda. comparison of the reactivity of the sera with extracts of vaccinia virus ...19902210882
identification of the atp-binding domain of vaccinia virus thymidine kinase.although small in size (20 kda), the vaccinia virus (vv) thymidine kinase protein (ec 2.7.1.21 tk) is a relatively complex enzyme which must contain domains involved in binding both substrates (atp and thymidine) and a feedback inhibitor (dttp), as well as sequences directing the association of individual protein monomers into a functional tetrameric enzyme. alignment of predicted amino acid sequences of the thymidine kinase genes from a variety of sources was used to identify highly conserved r ...19902211649
dna sequence of the gene encoding a major secreted protein of vaccinia virus, strain lister.infection of tissue culture cells with vaccinia virus results in the specific secretion of several polypeptides into the medium. previous studies identified a protein of approximate mr 35,000 (35k) which was secreted in large amounts at both early and late times after infection with the evans strain. we now show that a related protein is secreted by the lister strain but not by wr, wyeth nor tian tan. the gene encoding the lister strain 35k protein was mapped within the inverted terminal repeats ...19902212991
vaccinia virus gene encoding a 30-kilodalton subunit of the viral dna-dependent rna polymerase.antibody was raised against purified vaccinia virus rna polymerase and used to screen a recombinant vaccinia virus-lambda gt11 library. the dna from several immunopositive clones was shown by southern hybridization to originate from the vaccinia virus hindiii e fragment. the nucleotide sequence of the rna polymerase subunit gene predicts a polypeptide 287 amino acids in length and 30,000 daltons in mass. an early transcript with a 5' terminus just upstream of the putative initiation codon was id ...19902214020
transcription factors nfi and nfiii/oct-1 function independently, employing different mechanisms to enhance adenovirus dna replication.initiation of adenovirus dna replication is strongly enhanced by two transcription factors, nuclear factor i (nfi) and nuclear factor iii (nfiii/oct-1). these proteins bind to two closely spaced recognition sequences in the origin. we produced nfi and nfiii/oct-1, as well as their biologically active, replication-competent dna-binding domains (nfi-bd and the pou domain), in a vaccinia virus expression system and purified these polypeptides to apparent homogeneity. by dnase i footprinting and gel ...19902214023
nucleotide sequence analysis and expression from recombinant vectors demonstrate that the attachment protein g of bovine respiratory syncytial virus is distinct from that of human respiratory syncytial virus.bovine respiratory syncytial (brs) virus causes a severe lower respiratory tract disease in calves similar to the disease in children caused by human respiratory syncytial (hrs) virus. while there is antigenic cross-reactivity among the other major viral structural proteins, the major glycoprotein, g, of brs virus and that of hrs virus are antigenically distinct. the g glycoprotein has been implicated as the attachment protein for hrs virus. we have carried out a molecular comparison of the glyc ...19902214024
determination of influenza virus proteins required for genome replication.an artificial vaccinia virus vector-driven replication system for influenza virus rna has been developed. in this system, a synthetic ns-like gene is replicated and expressed by influenza virus proteins supplied through infection with vaccinia virus recombinant vectors. the minimum subset of influenza virus proteins needed for specific replication and expression of the viral ribonucleoprotein was found to be the three polymerase proteins (pb1, pb2, and pa) and the nucleoprotein.19902214032
virus dissemination via the lymphomyeloid complex.in the previous aids symposium organized by the society, witte and witte (1) made a number of predictions, one of which was that in aids patients, "lymph from the thoracic duct should be strongly positive for hiv." though direct evidence for this is lacking, some early experiments of ours with vaccinia virus (2) are fully in accord with this prediction, to which they lend indirect support. in rabbits, nasally instilled vaccinia virus spreads via the lymphatic pathway (afferent peripheral lymph-- ...19902214864
prevention of immunosuppression in stressed mice by neurotropin(nsp).it is well known that the immune function can be compromised by stress. to investigate immune function in mice stressed by experimental restraint or unavoidable and opioid dependent stress, we evaluated the changes in total body weight and in organ weights (liver, spleen and thymus) of these animals, as well as the phagocytic activity of macrophages, the cytotoxicity of t cells and inhibitory effects on tumor growth and changes in t cell subset populations. at the same time we evaluated the effe ...19902215070
iptg-dependent vaccinia virus: identification of a virus protein enabling virion envelopment by golgi membrane and egress.a novel method has been developed to study the functional roles of individual vaccinia virus gene products that is neither limited by the possible essentiality of the target gene nor by the availability of conditional lethal mutants. the system utilises the e. coli lac repressor protein, the operator sequence to which it binds and the specific inducer iptg. it allows the generation of recombinant viruses in which the expression of any chosen gene, and hence virus replication, can be externally c ...19902216706
sequence requirements for cytochromes p450iia1 and p450iia2 catalytic activity: evidence for both specific and non-specific substrate binding interactions through use of chimeric cdnas and cdna expression.cytochrome p450s iia1 and iia2, encoded by the cyp2a1 and cyp2a2 genes, display 88% amino acid sequence similarities. the dissimilarities of sequence between these two enzymes are primarily localized within four discrete regions of the polypeptides that are separated by regions of absolute sequence identity. iia1 specifically hydroxylates the prototype substrate testosterone at the 7 alpha and 6 alpha position with a predominance of 7 alpha metabolite. iia2, on the other hand, hydroxylates this ...19902217131
increased biological activity of a recombinant factor ix variant carrying alanine at position +1.in attempts to improve the post-translational modification and processing of recombinant factor ix (fix) we have altered the cdna sequence encoding pre-pro-fix using site-directed mutagenesis and have expressed the variant cdnas in bhk21 cells using a vaccinia-virus-derived vector. we find that substitution of the tyrosine residue at +1 for an alanine increases the biological activity of the recombinant molecules 2-fold. on the other hand, substitution of the proline at -3 for a valine results i ...19902217136
preferential activation and genotoxicity of food derived heterocyclic amine mutagens by recombinant cytochrome p4501a2. 19902217402
efficacy of a baiting system for vaccinating foxes against rabies with vaccinia-rabies recombinant virus.the efficacy of a vaccinia-rabies recombinant virus (10(8) tcid50) contained in a machine-made baiting system has been tested in 22 captive young foxes which were divided into three experimental groups of six and a control group of four foxes. each fox in groups 1, 2 and 3 were fed one, two and three vaccine-baits, respectively, on successive days. the four unvaccinated foxes were housed separately. as shown by the incorporation of a tetracycline biomarker into their bones, all the baited foxes ...19902219648
the vaccinia virus hindiii f fragment: nucleotide sequence of the left 6.2 kb.the nucleotide sequence of the left 6.2 kb of the 13.2-kb hindiii f fragment of vaccinia virus was determined. translation of the sequence revealed nine closely spaced, tandemly oriented open reading frames (orfs), all reading leftward. the transcriptional organization of this region was determined by northern blot and s1 nuclease mapping. the analysis suggested that orfs 1, 2, 4, 5, 6, 7, and 8 are transcribed early in infection, whereas orfs 3 and 9 are probably late genes. two of these orfs h ...19902219701
vaccinia virus gene d7r encodes a 20,000-dalton subunit of the viral dna-dependent rna polymerase.the polypeptide encoded by the vaccinia virus open reading frame d7r was synthesized in bacteria. immunization of rabbits with the polypeptide resulted in antibodies that specifically recognized a virion polypeptide of 20,000 daltons. the immunoreactivity with the 20,000-dalton polypeptide was found to coincide with the virion-associated dna-dependent rna polymerase through deae-cellulose chromatography and glycerol gradient sedimentation. these results argue that the product of the vaccinia ope ...19902219712
reverse guanine phosphoribosyltransferase selection of recombinant vaccinia viruses.we have developed a procedure for the selection of recombinant vaccinia viruses with applicability to poxvirus mutagenesis studies and to the use of vaccinia virus as an expression vector. the method depends on the specific inability of a recombinant vaccinia virus expressing the escherichia coli guanine phosphoriboxyltransferase gene (gpt) to form plaques on a hypoxanthine-guanine phosphoribosyltransferase-negative line of mouse fibroblasts in the presence of 6-thioguanine. recombinant viruses ...19902219714
the complete dna sequence of vaccinia virus.the complete dna sequence of the genome of vaccinia virus has been determined. the genome consisted of 191,636 bp with a base composition of 66.6% a + t. we have identified 198 "major" protein-coding regions and 65 overlapping "minor" regions, for a total of 263 potential genes. genes encoded by the virus were located by examination of dna sequence characteristics and compared with existing vaccinia virus mapping analyses, sequence data, and transcription data. these genes were found to be compa ...19902219722
a dna ligase gene in the copenhagen strain of vaccinia virus is nonessential for viral replication and recombination.biochemical and genetic analyses have been conducted to determine whether a vaccinia virus open reading frame (orf) with extensive homology to the saccharomyces cerevisiae dna ligase gene encodes a functional ligase activity. this orf in hindiii a, designated a50r, is capable of encoding a 552-amino-acid, 63.4-kda polypeptide. full-length a50r mrna produced in vitro directed the synthesis of a polypeptide with an apparent molecular weight of 57 kda. significantly, translation reactions programme ...19902219723
processing, assembly, and immunogenicity of human immunodeficiency virus core antigens expressed by recombinant vaccinia virus.recombinant vaccinia viruses that contained regions of the gag-pol open reading frames of human immunodeficiency virus type 1 (hiv-1) were constructed. cells infected with recombinants containing both gag and protease genes expressed and processed hiv gag antigens efficiently. processing was much reduced in cells infected with recombinants containing only gag, but not the protease gene. however, significant amounts of p41 were produced by protease-defective recombinants. this protein was immunor ...19902219727
the polypeptide composition of vaccinia-infected cell membranes and rifampicin bodies.the protein and glycoprotein composition of a sucrose gradient fraction from vaccinia infected cells treated with rifampicin was studied. this particulate fraction contained cytoplasmic membranes and pleomorphic membranous structures. the glycoproteins (89, 42 and 20-23 kda, respectively) were identified as the same glycoproteins that are found in plasma membranes of infected cells and the envelope of extracellular enveloped vaccinia (eev). these glycoproteins could be solubilized by 0.1% np-40. ...19902220070
priming effect of recombinant vaccinia virus coding for the middle hepatitis b surface antigen.recombinant vaccinia virus expressing the middle hepatitis b virus surface antigen was incapable of inducing marked antibody response against the s and pre-s2 antigenic specifities in mice. however, mice immunized with this virus produced antibodies to both these antigens after the following administration of subtreshold doses of plasmatic hepatitis b surface antigen.19902222186
[response in patients with melanoma to immunization using melanoma oncolysates of vaccine virus].thirty-two patients with high risk melanoma (either primary melanoma of the limbs or trunk, or recurrent melanoma) and clinically disease-free following appropriate surgical treatment were immunized with a vaccinia virus oncolysate made from a pool of 4 human melanoma cell lines. injections were given id weekly for 3 months, and then bi-monthly for a further 21 months or until relapse. treated patients have been under study for 11-72 months, and 15 of them for more than 36 months. twelve patient ...19902224160
immunotherapy of the rat 13762sc mammary adenocarcinoma by vaccinia virus augmentation of tumor immunity.we studied whether vaccinia virus (vv) functioned as an immunogenic carrier in augmenting anti-tumor immunity in rats bearing a syngeneic metastatic tumor. the primary tumor was induced by injecting 10(6) 13762sc mammary adenocarcinoma cells subcutaneously into the right hind footpad of fischer 344 rats. a concomitant anti-tumor response is induced by the tumor as demonstrated by the inhibited growth of a second tumor challenge given in the contralateral footpad 3-15 days later. attempts were ma ...19902225567
growth inhibition by vaccinia virus growth factor.vaccinia virus growth factor (vgf), a highly glycosylated 77-residue epidermal growth factor (egf)-like polypeptide encoded in vaccinia poxvirus, is reported to play an important role in stimulating growth of uninfected cells to facilitate virus infection. we have chemically synthesized the unglycosylated forms of vgf and vgf19-69, a shortened vgf analog consisting of 51 residues and comprising the egf-homologous region (position 19-69) of vgf. both synthetic forms of vgfs were purified to homog ...19902229050
inoculation of macaca fascicularis with simian immunodeficiency virus, sivmne immunologic, serologic, and pathologic changes.previous studies had tested the susceptibility of two macaque species, macaca nemestrina and m. mulatta, to infection with the primate lymphotropic lentivirus sivmne. in this report we describe the results obtained after infecting eleven m. fascicularis with sivmne. six of the animals had previously been immunized with a recombinant vaccinia virus expressing the envelope gene of hiv-1. all eleven animals became seropositive. to date ten animals have died 43 to 155 weeks post infection of an aids ...19902231689
estrogen receptor level determines sex-specific in vitro transcription from the xenopus vitellogenin promoter.female-specific expression of the xenopus laevis vitellogenin gene was reconstituted in vitro by addition of recombinant vaccinia-virus-produced estrogen receptor to nuclear extracts from male livers, in which this gene is silent. transcription enhancement was at least 30 times and was selectively restricted to vitellogenin templates containing the estrogen-responsive unit. thus, in male hepatocytes, estrogen receptor is the limiting regulatory factor that in the female liver controls efficient ...19902236004
inhibition of the complement cascade by the major secretory protein of vaccinia virus.the complement system contributes to host defenses against invasion by infectious agents. a 35-kilodalton protein, encoded by vaccinia virus and secreted from infected cells, has sequence similarities to members of a gene family that includes complement control proteins. biochemical and genetic studies showed that the viral protein binds to derivatives of the fourth component of complement and inhibits the classical complement cascade, suggesting that it serves as a defense molecule to help the ...19902237434
the human immunodeficiency virus type 1 envelope glycoprotein precursor acquires aberrant intermolecular disulfide bonds that may prevent normal proteolytic processing.the envelope glycoprotein of human immunodeficiency virus consists of two subunits, designated gp120 and gp41, derived from the cleavage of a precursor polypeptide gp160. when expressed from a recombinant vaccinia virus and analyzed by velocity gradient sedimentation and polyacrylamide gel electrophoresis, a significant proportion of gp160 molecules formed oligomers that were stabilized by intermolecular disulfide bonds. oligomeric forms of both gp120 and gp41 were also observed, but these oligo ...19902238472
recombinant vaccinia viruses as vectors for studying t lymphocyte specificity and function. 19902242679
posttranslational modification of vaccinia virus proteins. 19902242680
generation of hybrid genes and proteins by vaccinia virus-mediated recombination: application to human immunodeficiency virus type 1 env.the ability of poxviruses to undergo intramolecular recombination within tandemly arranged homologous sequences can be used to generate chimeric genes and proteins. genes containing regions of nucleotide homology will recombine to yield a single sequence composed of portions of both original genes. a recombinant virus containing two genes with a number of conserved regions will yield a population of recombinant viruses containing a spectrum of hybrid sequences derived by recombination between th ...19902243381
use of a cell-free system to identify the vaccinia virus l1r gene product as the major late myristylated virion protein m25.a 25-kda vaccinia virus (vv) virion protein, designated m25, is modified in vivo by covalent addition of myristic acid. the predicted amino acid sequences of all vv open reading frames which have been reported were searched for the sequence m-g-x-x-x-(s/t/a), which has been proposed to be the consensus recognition signal for cotranslational modification of proteins by n-myristyltransferase. this conserved signal was found at the amino terminus of a single locus, which corresponded to the leftmos ...19902243383
mutational analysis of a vaccinia virus intermediate promoter in vivo and in vitro.the expression of the vaccinia virus intermediate i3 gene depends on trans-acting factors which are present in an active state prior to dna replication. however, activation of transcription requires dna replication in cis (j. c. vos and h. g. stunnenberg, embo j., 7:3487-3492, 1988). we have made deletion and linker scanner mutations of the i3 promoter to determine the sequence requirements for transcriptional activity and the dependence of dna replication. the i3 promoter appears to consist of ...19902243387
influence of s-adenosylhomocysteine hydrolase inhibitors on s-adenosylhomocysteine and s-adenosylmethionine pool levels in l929 cells.s-adenosylhomocysteine hydrolase has been recognized as the target enzyme for the antiviral activity of several carbocyclic and acyclic adenosine analogues. in a previous study [cools m and de clercq e, biochem pharmacol 38: 1061-1067, 1989], we found a close correlation between the antiviral activity of six adenosine analogues [s)-9-(2,3-dihydroxypropyl)adenine [(s)-dhpa], (rs)-3-adenin-9-yl-2-hydroxypropanoic acid [(rs)-ahpa] (isobutyl ester), 3-deazaneplanocin a, carbocyclic 3-deazaadenosine ...19902244927
vaccinia recombinant virus expressing the rabies virus glycoprotein: safety and efficacy trials in canadian wildlife.twenty-six meadow voles (microtus pennsylvanicus), ten woodchucks (marmota monax), thirteen grey squirrels (sciurus carolinensis), thirteen ring-billed gulls (larus delawarensis), six red-tailed hawks (buteo jamaicensis) and eight great horned owls (bubo virginianus) received vaccinia virus recombinant expressing the rabies virus glycoprotein (v-rg) by direct instillation into the oral cavity. each of ten coyotes (canis latrans) received the virus in two vaccine-laden baits. several voles and mo ...19902249183
conditional expression of foreign genes by temperature-sensitive mutants of vaccinia virus.to assess the utility of two temperature-sensitive (ts) mutant vaccinia viruses as vectors for the conditional in vitro expression of recombinant foreign genes, we have studied the kinetics of expression of foreign genes incorporated into these viruses. at nonpermissive temperature, 40 degrees c, these viruses were defective either in dna synthesis or in virus assembly. foreign gene expression was affected by the nature of the ts lesion and by the nature of the vaccinia promoter positioned upstr ...19902249782
determination of functional effects of mutations in the steroid 21-hydroxylase gene (cyp21) using recombinant vaccinia virus.steroid 21-hydroxylase (p450c21) is absent or defective in more than 90% of patients with congenital adrenal hyperplasia. this disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (cyp21) have been found in association with particular clinical phenotypes. to determine the functional effects of mutations causing amino acid substitutions, normal p450c21 and three mutagenized p450c21 enzymes were expressed at high levels in ...19902249999
[molecular-biological study of vaccinia virus genome. i. cloning of vaccinia virus dna fragments in bacterial vectors].the hindiii dna fragments of vaccinia virus strain l-ivp were cloned in pbr322 bacterial plasmid. a hybrid plasmids collection of pvhn series contains all fragments of virus genome except terminal hindiii-b and hindiii-g, and also a large hindiii-a. the latter was cloned in cosmid phc79. the obtained collection of hybrid dna molecules allows to carry out a wide range of molecular biological experiments on the vaccinia virus genome.19902250684
[molecular-biological study of vaccinia virus genome. ii. localization and nucleotide sequence of vaccinia virus genes coding for proteins 36k and 12k].genes encoding virus-specific late proteins with molecular mass 36 kda and 12 kda were mapped in hindiii-p dna fragment of vaccinia virus strain l-ivp by hybrid selection of rna to cloned dna fragments followed by in vitro translation. rna origin site of the 36k protein was detected in hindiii-j fragment. nucleotide sequences of these genes were determined. amino acid sequences of the 36k and 12k polypeptides were compared with the protein bank pir.19902250685
[molecular-biological study of vaccinia virus genome. iii. identification of the late gene product protein 36k from hind-iii-p-fragment of vaccinia virus strain l-ivp].vaccinia virus gene encoding 36k protein was cloned in pur290 bacterial expressing vector and resulted in the synthesis of a chimeric protein in e. coli. the chimeric protein consists of beta-galactosidase and virus protein in c-termini. it has virus antigen specificity. by monospecific antibody 36k protein of vaccinia virus was determined to be non-virion. it is localized in the cytoplasm of infected cells.19902250686
vaccination against tumor cells expressing breast cancer epithelial tumor antigen.ninety-one percent of breast tumors aberrantly express an epithelial tumor antigen (eta) identified by monoclonal antibody h23. vaccinia virus recombinants expressing tumor antigens have considerable promise in the active immunotherapy of cancer, and we have evaluated the potential of vaccinia recombinants expressing the secreted (s) and cell-associated (transmembrane, t) forms of h23 eta to elicit immunity to tumor cells expressing eta. tumorigenic ras-transformed fischer rat fibroblast lines f ...19902251291
potential use of live viral and bacterial vectors for vaccines. who meeting, geneva, 19-22 june, 1989.the use of vaccinia virus vector for the delivery of antigens was first described by moss and paoletti and their colleagues in 1982. such vaccines could be of particular value in developing countries because they would be cheap, stable, easy to administer and provide long-lasting immunity. who recognized the potential value of such a delivery system by convening two meetings, one at the national institutes of health, usa in november 1984 and the second at who headquarters, geneva, switzerland in ...19902251869
biological characterization of recombinant vaccinia viruses in mice infected by the respiratory route.a murine model based on infection by the respiratory route has been used to study the pathogenesis of recombinant vaccinia viruses. the neurovirulent western reserve (wr) strain and the wyeth smallpox vaccine strain were used as vectors. recombinant viruses were constructed by insertion of the epstein-barr virus membrane glycoprotein 340 gene into the thymidine kinase (tk) gene of each vaccinia virus. intranasal inoculation of dba/2 mice with 10(6) pock-forming units (pk.f.u.) of the wr strain w ...19902254756
epidermal growth factor. 19902255900
[vaccinia virus stimulation of oncogenesis in c57bl mice]. 19902256321
expression of the structural proteins of dengue 2 virus and yellow fever virus by recombinant vaccinia viruses.vaccinia virus recombinants were constructed which contained cdna sequences encoding the structural region of dengue 2 virus (pr159/s1 strain) or yellow fever virus (17d strain). the flavivirus cdna sequences were expressed under the control of the vaccinia 7.5k early/late promotor. cultured cells infected with these recombinants expressed immunologically reactive flavivirus structural proteins, precursor prm and e. these proteins appeared to be cleaved and glycosylated properly since they comig ...19902260923
[a recombinant vaccinia virus expressing the middle pres2-s envelope protein of the hepatitis b virus. the induction of humoral and cellular immunity by the immunization of laboratory animals]. 19902265587
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