characterization of an r-binding site mediating the r-induced activation of the epstein-barr virus bmlf1 cells latently infected with epstein-barr virus, the switch from latency to productive infection is linked to the expression of two epstein-barr virus transcription factors called eb1 and r. r is an enhancer factor, and an r-responsive element (rre) has been identified in the bmlf1 promoter. in this study, we have used bidirectional deletion mutagenesis to delineate the bmlf1 rre (rre-m) to a 44-bp sequence. we also show that r expressed from a recombinant vaccinia virus protects rre-m agains ...19921309256
mhv s peplomer protein expressed by a recombinant vaccinia virus vector exhibits igg fc-receptor activity.we have previously shown that cells infected with mouse hepatitis virus (mhv) bind rabbit, mouse, and rat igg by the fc portion of the igg molecule. this fc-binding activity appeared to be mediated by the mhv s protein. s protein could also be precipitated from mhv-infected cells by a monoclonal antibody directed against the murine fc gamma receptor (fc gamma r). to prove definitively that the s protein mediates fc-binding activity, we have expressed the mhv s protein utilizing recombinant vacci ...19921309271
cell-specific posttranslational events affect functional expression at the plasma membrane but not tetrodotoxin sensitivity of the rat brain iia sodium channel alpha-subunit expressed in mammalian cells.the rat brain iia na+ channel alpha-subunit was expressed and studied in mammalian cells. cells were infected with a recombinant vaccinia virus (vv) carrying the bacteriophage t7 rna polymerase gene and were transfected with cdna encoding the iia na+ channel alpha-subunit under control of a t7 promoter. whole-cell patch-clamp recording showed that functional iia channels were expressed efficiently (approximately 10 channels/microns2 in approximately 60% of cells) in chinese hamster ovary (cho) c ...19921309573
cloning of the vaccinia virus ribonucleotide reductase small subunit gene. characterization of the gene product expressed in escherichia coli.during its infectious cycle, vaccinia virus expresses a virus-encoded ribonucleotide reductase which is distinct from the host cellular enzyme (slabaugh, m.b., and mathews, c.k. (1984) j. virol. 52, 501-506; slabaugh, m.b., johnson, t.l., and mathews, c.k. (1984) j. virol. 52, 507-514). we have cloned the gene for the small subunit of vaccinia virus ribonucleotide reductase (designated vvr2) into escherichia coli and expressed the protein using a t7 rna polymerase plasmid expression system. afte ...19921309792
virus-induced loss of class i mhc antigens from the surface of cells infected with myxoma virus and malignant rabbit fibroma virus.shope fibroma virus (sfv) is a leporipoxvirus that causes localized benign fibromas in immunocompetent adult rabbits that spontaneously regress due, in part, to a cell-mediated immune response. myxoma virus (myx) and malignant rabbit fibroma virus (mrv) are related leporipoxviruses that induce rapidly lethal generalized infections accompanied by tumors and immunosuppression. because only these latter two viruses are known to compromise cell-mediated antiviral responses, cell surface levels of cl ...19921309843
nuclear binding of purified retinoblastoma gene product is determined by cell cycle-regulated phosphorylation.the retinoblastoma tumor suppressor gene product (prb) is a nuclear protein subject to cell cycle-regulated hyperphosphorylation. i constructed a recombinant vaccinia virus vector that expresses both the underphosphorylated and hyperphosphorylated forms of prb and purified the recombinant protein by using immunoaffinity chromatography directed toward a synthetic carboxy-terminal epitope. to investigate the hypothesis that hyperphosphorylation of prb is a means of controlling its growth-regulatin ...19921310146
recombinant fowlpox viruses expressing the glycoprotein b homolog and the pp38 gene of marek's disease virus.two marek's disease virus (mdv) genes, one homologous to the glycoprotein b gene of herpes simplex virus and encoding the b antigen complex and the other encoding a 38-kda phosphorylated protein (pp38), were inserted into the fowlpox virus (fpv) genome under the control of poxvirus promoters. randomly selected nonessential regions of fpv were used for insertion, and the vaccinia virus 7.5 kda polypeptide gene promoter or a poxvirus synthetic promoter was used for expression of mdv genes. gene ex ...19921310754
functional interactions between the fusion protein and hemagglutinin-neuraminidase of human parainfluenza viruses.the fusion glycoprotein (f) and hemagglutinin-neuraminidase (hn) genes of human parainfluenza virus type 2 (pi2) were molecularly cloned and expressed in hela-t4 cells by using the vaccinia virus-t7 transient expression system. expression of the f and hn proteins was detected by using immunoprecipitation and surface immunofluorescence staining. although the f protein was found to be cleaved into f1 and f2 and expressed on cell surfaces, no cell fusion was observed. however, cotransfection of the ...19921310764
insertion of the human immunodeficiency virus cd4 receptor into the envelope of vesicular stomatitis virus particles.enveloped virus particles carrying the human immunodeficiency virus (hiv) cd4 receptor may potentially be employed in a targeted antiviral approach. the mechanisms for efficient insertion and the requirements for the functionality of foreign glycoproteins within viral envelopes, however, have not been elucidated. conditions for efficient insertion of foreign glycoproteins into the vesicular stomatitis virus (vsv) envelope were first established by inserting the wild-type envelope glycoprotein (g ...19921310767
human cd8+ herpes simplex virus-specific cytotoxic t-lymphocyte clones recognize diverse virion protein antigens.the role of the hla class i-restricted, cd8+, herpes simplex virus (hsv)-specific cytotoxic t lymphocytes (ctl) in the control of human hsv infections is controversial because previous reports suggest that a substantial portion of the antigen-specific lytic response is mediated by cd4+ cells. to address this question directly, we isolated hsv-specific cd8+ ctl clones from a patient with recurrent genital herpes. these ctl were cloned by coculturing responder peripheral blood mononuclear cells (p ...19921310769
nuclear expression of the lower matrix protein of human cytomegalovirus in peripheral blood leukocytes of immunocompromised viraemic patients.peripheral blood leukocytes (pbl), namely polymorphonuclear leukocytes (pmnl), are the major carrier of human cytomegalovirus (hcmv) in the blood of immunocompromised patients with hcmv viraemia. by using monoclonal antibodies (mabs) directed against different early and late viral proteins, we showed that the protein accumulating in pbl, originally reported to be an immediate early (ie) gene product, is the 65k lower matrix early structural protein (ep65). this protein is detectable by immunoflu ...19921311365
characterization of the varicella-zoster virus gene 61 protein.the protein predicted to be encoded by varicella-zoster virus (vzv) gene 61 exhibits limited amino acid sequence similarity to the herpes simplex virus type 1 nuclear phosphoprotein vmw110, which functions as a transcriptional activator. the gene 61 protein was expressed in its entirety, or as an amino- or carboxy-terminal fragment in escherichia coli and vaccinia virus recombinants, and monospecific rabbit antisera were raised against an e. coli fusion between beta-galactosidase and the majorit ...19921312115
effective vaccination against papilloma development by immunization with l1 or l2 structural protein of cottontail rabbit papillomavirus.immunization of rabbits with either l1, the major structural protein, or l2, a minor structural protein of cottontail rabbit papillomavirus (crpv), protected against challenge with the virus. neutralizing antibodies were elicited by both the l1 and l2 trpe fusion proteins. neutralization with anti-l1 serum, however, was more efficient than with anti-l2 serum. in contrast, when tested on western blots the immune response to l2 was stronger than to l1. rabbits were also protected against crpv infe ...19921312271
acute inflammatory response to cowpox virus infection of the chorioallantoic membrane of the chick embryo.the chick embryo chorioallantoic membrane was used to study the acute inflammatory response in the absence of contributions from the immune system. in preliminary experiments, lesions of wild-type cowpox virus strain brighton (cpv-br) and a 38k gene deletion mutant of cpv-br (cpv-br.d1) were compared with vaccinia virus (strains wr and copenhagen), fowlpox virus, laryngotracheitis virus, and infectious tenosynovitis virus, and were ranked for degree of induced inflammation. the maximal and minim ...19921312273
expression of cdna encoding the sendai virus hemagglutinin-neuraminidase gene: characterization of wild-type and mutant gene products.cloned cdna encoding the sendai virus (sv) hemagglutinin-neuraminidase (hn) envelope glycoprotein was expressed in cultured cells in two ways: (i) infection with hn-expressing recombinant vaccinia virus, or (ii) transfection with a plasmid with t7 promoter and termination sequences flanking the hn gene, with intracellular t7 rna polymerase supplied by coinfection with recombinant vaccinia virus that expresses the enzyme. the hn expressed was indistinguishable from the authentic sv protein in ant ...19921312281
a novel herpes simplex virus glycoprotein, gl, forms a complex with glycoprotein h (gh) and affects normal folding and surface expression of gh.a glycoprotein encoded by the ul1 gene of herpes simplex virus type 1 (hsv-1) was detected in infected cells with antipeptide sera. the ul1 gene has previously been implicated in virus-induced cell fusion (s. little and p. a. schaffer, virology 112:686-697, 1981). two protein species, a 30-kda precursor form and a 40-kda mature form of the glycoprotein, both of which were modified with n-linked oligosaccharides, were observed. this novel glycoprotein is the 10th hsv-1 glycoprotein to be describe ...19921312629
in vitro recognition of an orf virus early promoter in a vaccinia virus extract.dna fragments containing varying lengths of the 5' end of an orf virus early gene (orf3) and its associated promoter were introduced into sodium deoxycholate-solubilized vaccinia virus extracts capable of initiating transcription in vitro from vaccinia virus early promoters. after separation of the radiolabelled products of the reactions on a 5% polyacrylamide/7 m urea gel, discrete transcripts were detected the sizes of which were consistent with initiation of transcription from the orf virus e ...19921312824
tn9 cat gene contains a promoter for vaccinia virus transcription: implications for reverse-genetic techniques.vaccinia virus-dependent cat expression was observed in virus-infected cells cotransfected with a promoterless cat gene. restriction endonuclease resection of the cat plasmid indicated that expression was due to recognition by vaccinia virus rna polymerase of sequences within the cat gene itself, probably located within the 5' untranslated region of the gene. this observation is relevant to the design of reverse-genetic systems which use cat as a reporter gene to detect replication of negative-s ...19921313041
[virucidal effectiveness of some commercial products for chemothermal disinfection methods for temperature resistant viruses and bacteriophages--evaluation of a test model].in the laboratory assay of chemothermal virus disinfection procedures a test system is required which allows to differentiate between the physical effects of the temperature and the chemical effects by the disinfectant used. from the four test viruses recommended by the german association for the control of virus diseases (deutsche vereinigung zur bekämpfung der viruskrankheiten) only sv 40 tumor virus showed a sufficient thermal stability at 55 degrees c to take it in consideration as test viru ...19921313238
cap-specific mrna (nucleoside-o2'-)-methyltransferase and poly(a) polymerase stimulatory activities of vaccinia virus are mediated by a single protein.the vaccinia virus gene for s-adenosyl-l-methionine:mrna (nucleoside-o2'-)-methyltransferase, an enzyme required for the formation of the 5' cap structure of mrna, was identified. protein sequence analysis revealed that this cap-specific methyltransferase is derived from the same open reading frame as that previously shown to encode vp39, a mr 39,000 dissociable subunit of poly(a) polymerase that stimulates the formation of long poly(a) tails. consistent with this finding, methyltransferase acti ...19921313572
vaccinia-rotavirus vp7 recombinants protect mice against rotavirus-induced diarrhoea.recombinant vaccinia viruses expressing wild type intracellular vp7 (vp7wt) from rotavirus sa11 or vp7sc, a cell surface-anchored variant, boosted antibody titres in sa11-immune mice. pups born to these mice were protected from diarrhoea following challenge with sa11. in rotavirus-naive mice, two immunizations with recombinant vaccinia virus expressing vp7sc stimulated protective immunity that could be transferred to pups, whereas viruses expressing vp7wt did not stimulate protective immunity. r ...19921313627
expression of aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus: self-assembly of capsid proteins into particles.a portion of a cdna clone containing coding sequences for both structural proteins (vp1 and vp2) of aleutian mink disease parvovirus (adv) was inserted into recombinant vaccinia viruses, vv:adsp. immunohistochemical staining of vv:adsp-infected cells revealed that the adv antigen was readily detected and localized in the nuclei of infected cells. analysis of vv:adsp-infected cell lystates indicated that both vp1 and vp2 were produced and comigrated with authentic vp1 and vp2 from adv-infected cr ...19921313919
dna strand transfer reactions catalyzed by vaccinia topoisomerase i.vaccinia virus dna topoisomerase i forms a 3'-phosphoryl intermediate with duplex dnas containing the conserved binding/cleavage motif 5'ccctt decreases. covalently bound enzyme is capable of transferring the incised dna strand to a heterologous dna acceptor containing a 5'oh terminus. both intramolecular and intermolecular religation reactions are catalyzed. intramolecular strand transfer occurs to the noncleaved strand of the dna duplex and results in formation of a hairpin loop. intermolecula ...19921314832
fusogenic segments of bovine leukemia virus and simian immunodeficiency virus are interchangeable and mediate fusion by means of oblique insertion in the lipid bilayer of their target cells.modified bovine leukemia virus (blv) glycoproteins were expressed by using vaccinia virus recombinants, and their fusogenic capacities were examined by a syncytia-formation assay. this analysis indicates that (i) both blv envelope glycoproteins gp51 and gp30 are necessary for cell fusion; (ii) insertion of the n-terminal segment of gp30 (fusion peptide) into the lipid bilayer in an oblique orientation, as predicted by computer conformational analysis, results in fusogenic capacities higher than ...19921315040
protection of mice and swine from pseudorabies virus conferred by vaccinia virus-based recombinants.glycoproteins gp50, gii, and giii of pseudorabies virus (prv) were expressed either individually or in combination by vaccinia virus recombinants. in vitro analysis by immunoprecipitation and immunofluorescence demonstrated the expression of a gii protein of approximately 120 kda that was proteolytically processed to the giib (67- to 74-kda) and giic (58-kda) mature protein species similar to those observed in prv-infected cells. additionally, the proper expression of the 90-kda giii and 50-kda ...19921316458
transient expression and mutational analysis of the rotavirus intracellular receptor: the c-terminal methionine residue is essential for ligand binding.maturation of rotavirus involves an intracellular membrane budding event in which the single-shelled icosahedral particle interacts with a virus-encoded receptor glycoprotein, ns28, that is located in the rough endoplasmic reticulum membrane. the receptor is a tetramer and is oriented with the c-terminal 131 amino acids on the cytoplasmic side of the membrane (a.r. bellamy and g.w. both, adv. virus res. 38:1-48, 1990). we have used the t7-vaccinia virus transient expression system to deliver mut ...19921316468
protein of macaques against infection with simian type d retrovirus (srv-1) by immunization with recombinant vaccinia virus expressing the envelope glycoproteins of either srv-1 or mason-pfizer monkey virus (srv-3).rhesus macaques were immunized with live vaccinia virus recombinants expressing the envelope glycoproteins (gp70 and gp22) of simian type d retrovirus (srv), serotype 1 or 3. all of the animals immunized with either the srv-1 env or the srv-3 env vaccinia virus recombinant developed neutralizing antibodies against the homologous srv. in addition, both groups developed cross-reactive antibodies and were protected against an intravenous live-virus challenge with srv-1. the four control animals imm ...19921316495
molecular cloning and expression of a spike protein of neurovirulent murine coronavirus jhmv variant cl-2.a cdna encoding the spike (s) protein of the neurovirulent murine coronavirus jhmv variant cl-2 was isolated and sequenced. analysis of the cdna revealed that the s protein consists of 1376 amino acids, as does the s protein of mouse hepatitis virus 4. we inserted the cdna into the genome of vaccinia virus to obtain a recombinant vaccinia virus (rvv). the s protein expressed in rk13 cells infected by the rvv was shown to be electrophoretically and immunologically indistinguishable from the s pro ...19921316938
recent advances in antitumor vaccines.immunization with anti-idiotypic antibodies can induce cell-mediated and humoral antitumor immunity in animal models. this immunity can sometimes cause tumor destruction. however, more needs to be learned about how best to induce the type of immune response that is responsible for tumor destruction, since the presence of anti-idiotypic antibodies has been shown occasionally to enhance, rather than to inhibit, tumor growth. there is evidence suggesting that immunization of human cancer patients w ...19921318138
binding of the coronavirus mouse hepatitis virus a59 to its receptor expressed from a recombinant vaccinia virus depends on posttranslational processing of the receptor glycoprotein.recently, we showed that a murine member of the carcinoembryonic antigen family of glycoproteins serves as a cellular receptor (mhvr) for the coronavirus mouse hepatitis virus a59 (mhv-a59) (g. s. dveksler, m. n. pensiero, c. b. cardellichio, r. k. williams, g.-s. jiang, k. v. holmes, and c. w. dieffenbach, j. virol. 65:6881-6891, 1991; r. k. williams, g.-s. jiang, and k. v. holmes, proc. natl. acad. sci. usa 88:5533-5536, 1991). to examine the role of posttranscriptional modification of mhvr on ...19921318394
identification of the murine cytomegalovirus glycoprotein b gene and its expression by recombinant vaccinia virus.the gene encoding glycoprotein b (gb) of murine cytomegalovirus (mcmv) strain smith was identified, sequenced, and expressed by recombinant vaccinia virus. the gb gene was found adjacent to the polymerase gene, as it is in the genome of human cytomegalovirus (hcmv). the open reading frame consists of 2,784 nucleotides capable of encoding a protein of 928 amino acids. comparison with gb homologs of other herpesviruses revealed a high degree of homology. the similarity between the mcmv gb and the ...19921318410
myristylation of poliovirus capsid precursor p1 is required for assembly of subviral particles.the poliovirus capsid precursor polyprotein, p1, is cotranslationally modified by the addition of myristic acid. we have examined the importance of myristylation of the p1 capsid precursor during the poliovirus assembly process by using a recently described recombinant vaccinia virus expression system which allows the independent production of the poliovirus p1 protein and the poliovirus 3cd proteinase (d. c. ansardi, d. c. porter, and c. d. morrow, j. virol. 65:2088-2092, 1991). we constructed ...19921318418
active specific immunotherapy with vaccinia melanoma oncolysate.based on antitumor effects observed with vaccinia virus infected tumor cell lysate in animal models, adjuvant immunotherapeutic clinical trials were undertaken in patients with melanoma using vaccinia virus infected melanoma oncolysate (vmo). preliminary clinical trials showed that the vmo is safe except minimal side effects such as mild fever, pain, and tenderness at the site of vmo injection, and mild lymphadenopathy. the effective dose of vmo was investigated in a following trial using 0.05 t ...19921318507
identification of target antigens for the human cytotoxic t cell response to epstein-barr virus (ebv): implications for the immune control of ebv-positive malignancies.epstein-barr virus (ebv), a human herpes virus with oncogenic potential, persists in b lymphoid tissues and is controlled by virus-specific cytotoxic t lymphocyte (ctl) surveillance. on reactivation in vitro, these ctls recognize ebv-transformed lymphoblastoid cell lines (lcls) in an hla class i antigen-restricted fashion, but the viral antigens providing target epitopes for such recognition remain largely undefined. here we have tested ebv-induced polyclonal ctl preparations from 16 virus-immun ...19921319456
(+-)-carbocyclic 5'-nor-2'-deoxyguanosine and related purine derivatives: synthesis and antiviral properties.beginning with 3-cyclopenten-1-ylamine hydrochloride, the 5'-nor derivatives of carbocyclic 2'-deoxyguanosine (2), 2'-deoxyadenosine (3), and 2,6-diaminopurine 2'-deoxyribofuranoside (4) have been prepared. these compounds were evaluated for antiviral potential versus herpes simplex virus, varicella-zoster virus, cytomegalovirus, vaccinia virus, vesicular stomatitis virus, and human immunodeficiency virus and found to lack activity. also, compounds 2-4 were virtually nontoxic toward the host (hu ...19921319491
complexes of sendai virus np-p and p-l proteins are required for defective interfering particle genome replication in vitro.we present evidence that the formation of np-p and p-l protein complexes is essential for replication of the genome of sendai defective interfering (di-h) virus in vitro, using extracts of cells expressing these viral proteins from plasmids. optimal replication of di-h nucleocapsid rna required extracts of cells transfected with critical amounts and ratios of each of the plasmids and was three- to fivefold better than replication with a control extract prepared from a natural virus infection. ex ...19921321276
a novel glycoprotein of feline infectious peritonitis coronavirus contains a kdel-like endoplasmic reticulum retention signal.a new protein of feline infectious peritonitis coronavirus (fipv) was discovered in lysates of [35s]cysteine-labeled infected cells. expression of open reading frame (orf) 6b of fipv in recombinant vaccinia virus-infected cells was used to identify it as the 6b protein. further characterization revealed that it is a novel type of viral glycoprotein whose function is not clear. it is a soluble protein contained in microsomes; its slow export from the cell is caused by the presence of an endoplasm ...19921321279
immunity in strain 2 guinea-pigs inoculated with vaccinia virus recombinants expressing varicella-zoster virus glycoproteins i, iv, v or the protein product of the immediate early gene 62.the immunogenicity of specific varicella-zoster virus (vzv) proteins, with emphasis upon cell-mediated immune responses, was evaluated by immunizing strain 2 guinea-pigs with vaccinia virus recombinants that express gpi (vac-gpi), gpiv (vac-gpiv) and gpv (vac-gpv) or the ie-62 protein (vac-ie-62). vac-gpi elicited the highest initial mean t cell proliferation response [stimulation index (s.i.) 3.8 +/- 0.9 s.e.m.] whereas inoculation with vac-gpv produced the lowest primary t cell response (s.i. ...19921321876
vaccinia virus encodes four putative dna and/or rna helicases distantly related to each analysis of the amino acid sequences of vaccinia virus proteins containing the purine ntp-binding pattern revealed the seven motifs typical of the dna (rna) helicase superfamily ii in the proteins i8r and a18r. together with the previously described putative helicases d6r and d11l, the number of putative helicases of this superfamily encoded by the genome of vaccinia virus now reaches four. aside from the helicase motifs, the sequences of i8r and a18r showed no strong similarit ...19921321883
domain structure of human 72-kda gelatinase/type iv collagenase. characterization of proteolytic activity and identification of the tissue inhibitor of metalloproteinase-2 (timp-2) binding regions.the 72-kda gelatinase/type iv collagenase, a metalloproteinase thought to play a role in metastasis and in angiogenesis, forms a noncovalent stoichiometric complex with the tissue inhibitor of metalloproteinase-2 (timp-2), a potent inhibitor of enzyme activity. to define the regions of the 72-kda gelatinase responsible for timp-2 binding, a series of nh2- and cooh-terminal deletions of the enzyme were constructed using the polymerase chain reaction technique. the full-length and the truncated en ...19921322396
covalent and noncovalent dna binding by mutants of vaccinia dna topoisomerase i.analysis of vaccinia topoisomerase mutants that are impaired in dna relaxation has allowed the identification of amino acid residues required for the transesterification step of catalysis. missense mutations of wild-type residues gly-132----asp and arg-223----gln rendered the protein inert in formation of the covalent enzyme-dna complex and hence completely inactive in dna relaxation. mutations of thr-147----ile and gly-132----ser caused severe defects in covalent adduct formation that correlate ...19921322412
rotavirus vp6 modified for expression on the plasma membrane forms arrays and exhibits enhanced immunogenicity.the major inner capsid protein of rotavirus is vp6, a 42-kda polypeptide that forms the icosahedral surface of the rotavirus single-shelled particle. a chimeric form of vp6 (vp6sc) was constructed containing an upstream leader sequence derived from the influenza virus hemagglutinin and a downstream membrane-spanning (anchor) domain from a mouse immunoglobulin gene. when vp6sc was expressed in cells using a recombinant vaccinia virus, the protein was transported, glycosylated, and anchored in the ...19921322583
methyltransferase and subunit association domains of vaccinia virus mrna capping enzyme.rna triphosphatase, rna guanylyltransferase, and rna (guanine-n7-)-methyltransferase activities are associated with the vaccinia virus mrna capping enzyme, a heterodimeric protein containing polypeptides of m(r) 95,000 and 31,000. although the rna triphosphatase and rna guanylyltransferase domains have been localized to a m(r) 59,000 fragment of the capping enzyme large subunit, the location of the methyltransferase domain within the protein and the catalytic role of individual subunits in methy ...19921322901
the vaccinia virus mrna (guanine-n7-)-methyltransferase requires both subunits of the mrna capping enzyme for activity.plasmid vectors capable of expressing the large and small subunits of the vaccinia virus mrna capping enzyme were constructed and used to transform escherichia coli. conditions for the induction of the dimeric enzyme or the individual subunits in a soluble form were identified, and the capping enzyme was purified to near homogeneity. proteolysis of the capping enzyme in bacteria yields a 60-kda product shown previously to possess the mrna triphosphatase and guanyltransferase activities (shuman, ...19921322902
further characterization of the biological and pathogenic properties of erythromelalgia-related poxviruses.six isolates of erythromelalgia-related poxvirus (erpv) were characterized with respect to host range, c.p.e. and inclusions, pock formation on chorioallantoic membrane (cam), morphogenesis, serological reactivity, pathogenesis in animals and dna restriction fragment profile. the results suggest that erpv is either a new member of the orthopoxvirus genus or a subspecies of ectromelia virus. evidence is provided that (i) erpv has a wide host range in vitro in which characteristic viral c.p.e. and ...19921322958
overexpression of receptors of the steroid/thyroid family. 19921323054
side-chain derivatives of biologically active nucleosides. 1. side-chain analogs of 3'-azido-3'-deoxythymidine (azt).starting from 3-o-mesyl-1,2-o-isopropylidene-alpha-d-allofuranose (9) the anomeric mixtures of the requisite carbohydrates 1,2-di-o-acetyl-6-o-benzoyl-5-deoxy-3-o-mesyl-d-allofuranoses++ + 17a alpha/beta, 1,2-di-o-acetyl-5,6-di-o-benzoyl-3-o-mesyl-d-allofuranoses 17b alpha/beta, and 1,2-di-o-acetyl-5,6-di-o-benzoyl-3-o-mesyl-l-talofuranoses 17c alpha/beta were synthesized. 1,2-di-o-acetyl-5-o-benzoyl-6-deoxy-3-o-mesyl-d-allofuranoses++ + 17d alpha/beta and the corresponding l-talofuranoses 17e a ...19921323681
two classes of dna end-joining reactions catalyzed by vaccinia topoisomerase i.the ability of a eukaryotic dna topoisomerase i to catalyze dna rearrangements was examined in vitro using defined substrates and purified enzyme. site-specific dna strand cleavage by vaccinia topoisomerase i across from a nick generated double-strand breaks that could be religated to a heterologous blunt-ended duplex dna regardless of the sequence of the acceptor molecule. topoisomerase bound covalently at internal positions could religate the bound strand to an incoming acceptor provided that ...19921324909
analysis of the amino terminal presequence of the feline immunodeficiency virus glycoprotein: effect of deletions on the intracellular transport of gp95.the envelope glycoprotein of feline immunodeficiency virus (fiv) consists of two noncovalently associated subunits, the surface glycoprotein (su; gp95) and the transmembrane glycoprotein (tm; gp40). an unusual feature of the open reading frame (orf) encoding the fiv glycoprotein is the presence of an unusually long amino terminal sequence (149 amino acids, "l" region or n-region of the signal sequence) preceding the predicted hydrophobic signal sequence. to examine the role of this n-region in t ...19921325696
transmembrane signaling by the human insulin receptor kinase. relationship between intramolecular beta subunit trans- and cis-autophosphorylation and substrate kinase examine the role of intramolecular beta subunit trans- and cis-autophosphorylation in signal transduction, the vaccinia virus/bacteriophage t7 expression system was used to generate insulin holoreceptors composed of a kinase-defective half-receptor precursor (alpha beta a/k or alpha beta a/ ct) and a kinase-active half-receptor precursor (alpha beta delta ct or alpha beta wt). in the alpha beta a/k-alpha beta delta ct hybrid insulin receptor, insulin stimulated a 20-fold increase in in ...19921326556
synthesis and antiviral properties of (+/-)-5'-noraristeromycin and related purine carbocyclic nucleosides. a new lead for anti-human cytomegalovirus agent design.(+/-)-5'-noraristeromycin (3) has been prepared in three steps beginning with the 2,3-o-isopropylidene derivative of (+/-)-(1 alpha, 2 beta, 3 beta, 4 alpha)-4-amino-1,2,3-cyclopentanetriol (7). also prepared from the same starting material were the related hypoxanthine (4), guanine (5), and 2,6-diaminopurine (6) analogues. compounds 3-6 were evaluated for antiviral activity against a large number of viruses with marked activity being observed for 3 towards vaccinia virus, human cytomegalovirus, ...19921326633
a highly attenuated host range-restricted vaccinia virus strain, nyvac, encoding the prm, e, and ns1 genes of japanese encephalitis virus prevents jev viremia in swine.a highly attenuated strain of vaccinia virus (nyvac) was engineered to express the japanese encephalitis virus (jev) prm, e, and ns1 genes or the prm and e genes. the recombinant viruses were tested as vaccine candidates in pigs, a natural host of jev. jev-neutralizing and hemagglutination-inhibiting antibodies appeared in swine sera 7 days after immunization with 10(8) pfu of the recombinant viruses and increased after a second dose at 28 days. the jev levels detected in the serum after jev cha ...19921326813
gene translocations in poxviruses: the fowlpox virus thymidine kinase gene is flanked by 15 bp direct repeats and occupies the locus which in vaccinia virus is occupied by the ribonucleotide reductase large subunit sequencing a fragment of 7351 bp the fowlpox virus thymidine kinase gene has been found to map to a position within the equivalent of the vaccinia hindiii i fragment. the deduced gene arrangement in fowlpox virus is i3, x, tk, i5, i6, i7, i8, g1, indicating that the homologue of the vaccinia i4 gene has been replaced by two genes x and tk. the non-essential tk gene has therefore replaced another non-essential gene, i4 (the ribonucleotide reductase large subunit) in this region. the x/tk inser ...19921326827
conditional growth of escherichia coli caused by expression of vaccinia virus dna topoisomerase vaccinia virus topoisomerase i is expressed in escherichia coli containing plasmid p1940 (s. shuman, m. golder, and b. moss, j. biol. chem. 263:16401-16407, 1988). growth curves showed a decline of 2 to 3 logs in the number of viable cells at 42 degrees c after shift from 30 degrees c because of increased vaccinia virus topoisomerase i level. mutations in the gyra and gyrb genes allowed cells to grow equally well at 42 and 30 degrees c. the presence of gyrase inhibitor also improved growt ...19921328167
comparison of the locations of homologous fowlpox and vaccinia virus genes reveals major genome reorganization.we have derived a restriction enzyme map for the fowlpox virus fp9 strain. sites for bamhi, pvuii, psti and ncoi have been mapped mainly by southern blotting. the size of the genome derived from the restriction maps (254 kb) corresponds to the figure of 260 +/- 8 kb determined from analysis of genomic dna by pulsed-field electrophoresis. the map can be compared with a previously published map for a different strain of fowlpox virus using the psti digest which is common to both studies. some 65 k ...19921328478
proteolytic processing of a murray valley encephalitis virus non-structural polyprotein segment containing the viral proteinase: accumulation of a ns3-4a precursor which requires mature ns3 for efficient processing.the proteolytic processing of a non-structural polyprotein segment from the cytoplasmic domain of ns2a to the c terminus of ns5 of murray valley encephalitis (mve) virus was examined, when expressed from cdna via a vaccinia virus recombinant, in transiently transfected cos cells, or synthesized by cell-free translation. cleavages mediated by the virus-encoded proteinase domain in ns3 at the junctions of ns2a-2b, ns2b-3 and ns4b-5 were catalysed efficiently. however, the cleavage at the ns3-4a ju ...19921328486
expression and characterization of glycoprotein gp35 of hepatitis c virus using recombinant vaccinia virus.complementary dna clones corresponding to one of the putative structural regions of the hepatitis c virus (hcv) genome were obtained from sera of non-a non-b hepatitis patients. the putative envelope gene was expressed by using a recombinant vaccinia virus carrying this region of the hcv genome. in cells infected with the recombinant vaccinia virus, a glycosylated protein with an m(r) of about 35k (gp35) was specifically detected by convalescent sera from hepatitis c patients. the sera from rabb ...19921328487
herpes simplex virus-infected human fibroblasts are resistant to and inhibit cytotoxic t-lymphocyte activity.we examined the ability of human anti-herpes simplex virus (hsv) cytotoxic t lymphocytes (ctl) to lyse autologous human fibroblasts infected with hsv. in contrast to hsv-infected human epstein-barr virus-transformed b cells (lcl), which were lysed by hla-restricted anti-hsv ctl, autologous fibroblasts infected with hsv were resistant to lysis. this resistance was not due to a lack of infectivity or production of hsv proteins since greater than 90% of the cells were infected and expressed abundan ...19921328666
involvement of membrane traffic in the replication of poliovirus genomes: effects of brefeldin a.brefeldin a (bfa) is a macrolide antibiotic that has multiple targets in vesicular transport and blocks membrane traffic between the cis- and trans-golgi compartments, leading to the disruption of the trans-golgi apparatus (for a review see pelham, 1991, cell 67, 449-451). consequently, bfa interferes with the maturation of viral glycoproteins and suppresses the formation of infectious viruses that contain a lipid envelope. we report that this antibiotic strongly inhibits poliovirus replication ...19921329315
molecular cloning of natural paramyxovirus copy-back defective interfering rnas and their expression from dna.using the unique sequence organization of copy-back defective interfering (di) rnas of paramyxoviruses, sendai virus (sv), and measles virus copy-back di rnas were pcr amplified and cloned, without having to separate them from their helper nondefective genomes. the cloning was designed so that t7 polymerase transcription of the plasmids would generate di rnas with the exact 5' and 3' ends. the sv di clone, transcribed from the plasmid in bhk cells using t7 polymerase produced by a vaccinia virus ...19921329337
quantitative assessment of poxvirus promoters in fowlpox and vaccinia virus recombinants.a comparison was undertaken of poxvirus promoters in vaccinia and fowlpox virus (fpv) recombinants using the level of beta-galactosidase expressed from the lacz gene as a measure of promoter function. in this study a comparison was made of the vaccinia virus promoters, p 7.5 and p l11, the major late promoter of cowpox virus, p cpx (expressing the abundant inclusion body protein), and the fpv promoters, p e/l and p l. in vaccinia virus recombinants the fpv p e/l promoter expressed one-third to o ...19921329341
the soluble form of two n-terminal domains of the poliovirus receptor is sufficient for blocking viral means of deleting a c-terminal portion of the open reading frame of the poliovirus receptor cdna, and by vaccinia virus-mediated overexpression we have produced a protein corresponding to the first two n-terminal ig-like domains of the poliovirus receptor. this protein that lacked the third ig-like domain, the transmembrane region and most of the intracellular c-terminal tail was detected in the medium of vaccinia virus infected cells. the properties of the truncated pvr cdna were further cha ...19921329376
restimulated memory tc cells have a higher apparent avidity of interaction with targets than primary virus-immune tc cells as indicated by anti-cd8 blocking.previous experiments have shown that whereas a secondary in vitro kunjin-immune cytotoxic t (tc) cell population lysed equally well targets infected with either native flavivirus or a recombinant vaccinia virus expressing the immunodominant determinant, primary in vivo kunjin-immune tc cells were able to lyse only the recombinant vaccinia virus-infected targets. using cd8 blockade to assess the avidity of t cell-target interaction, recombinant-infected targets express antigen more efficiently th ...19921330899
nucleotide sequence of 21.8 kbp of variola major virus strain harvey and comparison with vaccinia virus.a 21.8 kbp region of the genome of variola major virus (strain harvey), a virus that caused haemorrhagic-type smallpox, has been sequenced and shown to possess 96% nucleotide identity to the corresponding region of vaccinia virus, the smallpox vaccine. overall the gene arrangement in the two viruses is highly similar and individual open reading frames (orfs) display a high degree of amino acid identity, for instance 26 of the 32 variola virus orfs have > or = 90% identity with their vaccinia vir ...19921331292
nucleotide sequence analysis of a unique near-terminal region of the tumorigenic poxvirus, shope fibroma virus.shope fibroma virus (sfv), a tumorigenic poxvirus, has a dna genome of approximately 160 kb. previous dna sequence analysis of sfv has been mainly limited to the terminal inverted repetitions (about 12 kb at each end of the genome) and immediately adjacent regions. we have sequenced a 4 kb fragment located approximately 20 kb from the right-terminal hairpin. within this region three complete and two partial open reading frames (orfs) have been identified. each of the putative polypeptides has se ...19921331293
a chimeric avian retrovirus containing the influenza virus hemagglutinin gene has an expanded host range.we have investigated what protein sequences are necessary for glycoprotein incorporation into rous sarcoma virus (rsv) virions by utilizing the hemagglutinin (ha) protein of influenza virus. two chimeric ha genes were constructed. in the first the coding sequence for the signal peptide of the rsv env gene product was fused in frame to the entire ha structural gene, and in the second the hydrophobic anchor and cytoplasmic domain sequences of the ha gene were also replaced with those from the rsv ...19921331528
the epstein-barr virus (ebv) nuclear antigen 1 bamhi f promoter is activated on entry of ebv-transformed b cells into the lytic epstein-barr virus (ebv)-positive burkitt's lymphoma cell lines exhibiting the latency i form of infection (i.e., ebv nuclear antigen 1 [ebna1] positive in the absence of other latent proteins), the ebna1 mrna has a unique bamhi q/u/k splice structure and is expressed from a novel promoter, fp, located near the bamhi fq boundary. this contrasts with the situation in ebv-transformed lymphoblastoid cell lines (lcls) exhibiting the latency iii form of infection (i.e., positive for all latent pro ...19921331531
independent evolution of monkeypox and variola viruses.smallpox was eradicated more than 10 years ago, but infection with another orthopoxvirus, monkeypox virus, can result in a clinical picture resembling smallpox. human infection with monkeypox virus is extremely rare, not easily transmitted, and confined to the rain forest belt of africa (z. jezek and f. fenner, p. 81-102, in human monkeypox, 1988). evidence that variola virus, the causative agent of smallpox, might be readily derived from monkeypox virus was presented [s. s. marennikova and e. m ...19921331540
attenuated deletion mutant of vaccinia virus ihd-w recovered virulence by reinsertion of a terminal restriction fragment.since vaccinia virus is being considered as a potential immunization vector, it is important to identify genes responsible for pathogenicity. one approach to identify virulence genes is the use of attenuated deletion mutants with a null background to reintroduce regions of the deleted genome material. we have previously described an attenuated deletion mutant of vaccinia virus strain ihd-w (z-19) that lost 21 kb from the left terminus comprising fragments hindiii c and n, displays rearrangements ...19921331699
vaccinia virus rna helicase: an essential enzyme related to the de-h family of rna-dependent ntpases.three distinct nucleic acid-dependent atpases are packaged within infectious vaccinia virus particles; one of these enzymes (nucleoside triphosphate phosphohydrolase ii or nph-ii) is activated by single-stranded rna. purified nph-ii is now shown to be an ntp-dependent rna helicase. rna unwinding requires a divalent cation and any one of the eight common ribo- or deoxyribonucleoside triphosphates. the enzyme acts catalytically to displace an estimated 10-fold molar excess of duplex rna under in v ...19921332061
characterization and purification of human retinoic acid receptor-gamma 1 overexpressed in the baculovirus-insect cell system.the full-length cdna for the human retinoic acid receptor-gamma 1 (rar-gamma 1) has been expressed to high levels in spodoptera frugiferda (sf9) cells using the baculovirus expression system. western blot analysis revealed that rar-gamma 1 expression increased between 32 and 60 h post-infection. the recombinant receptor was expressed primarily as a nuclear protein and displayed a molecular mass of 50 kda as determined by sds/page and gel-filtration chromatography, consistent with its cdna-deduce ...19921332684
an engineered mutant of vaccinia virus dna topoisomerase i is sensitive to the anti-cancer drug camptothecin.although highly homologous to the other eukaryotic type i dna topoisomerases, vaccinia virus dna topoisomerase i is distinct in its resistance to the anti-cancer drug camptothecin. after comparison of available sequences of sensitive and resistant type i topoisomerases, the aspartic acid at position 221 of vaccinia virus topoisomerase i is mutated to a valine. the resulting mutant protein is partially active. in contrast to the wild type enzyme, the relaxation of supercoiled dna is inhibited by ...19921332948
identification and functional analysis of the fowlpox virus homolog of the vaccinia virus p37k major envelope antigen gene.a fowlpox virus (fpv) gene with homology to the vaccinia virus p37k major envelope antigen gene was identified and sequenced. the predicted product has a molecular weight of 43,018 da (p43k). the fpv p43k gene has 37.5% identity with its vaccinia counterpart and higher homology with a molluscum contagiosum virus gene (42.6% identity). based on upstream sequences, p43k appears to be regulated as a late gene. recombinant fpv were generated in which a large portion of p43k was replaced by the esche ...19921333124
a yeast protein phosphatase related to the vaccinia virus vh1 phosphatase is induced by nitrogen starvation.a phosphatase related to the vaccinia virus vh1 phosphatase has been cloned from saccharomyces cerevisiae. the yeast phosphatase is related to the schizosaccharomyces pombe cdc25 gene product and to a protein encoded by a mammalian open reading frame known as 3ch134, which is an immediate early gene responding to serum stimulation. the phosphatase activity of the yeast gene product appears to be restricted to the hydrolysis of phosphotyrosine-containing substrates, whereas the vaccinia phosphata ...19921334559
synthesis and antiviral activity of some new s-adenosyl-l-homocysteine derivatives.a series of new s-adenosyl-l-homocysteine (adohcy) analogues with modifications to amino acid and nucleoside moieties was prepared via condensation of appropriate nucleoside precursors and suitably protected l-homocystine derivatives. the adohcy derivatives as well as the nucleoside precursors were evaluated for their antiviral activity. some of the compounds, in particular s-tubercidinyl-l-homocysteine propyl ester (36), n-(trifluoroacetyl)-s-tubercidinyl-l-homocysteine isopropyl ester (27), s- ...19921335077
possible approaches to develop vaccines against hepatitis a.more than a decade ago, successful replication of hepatitis a virus (hav) in cell culture opened the way to the development of live attenuated and inactivated vaccine candidates. serial passages of hav in cell culture led to attenuation as demonstrated by experiments in non-human primates. several live vaccine candidates obtained through serial passages have been evaluated in volunteers. significant improvements in the yield of viral antigen from infected cell cultures stimulated the development ...19921335659
antiviral (rna) activity of selected amaryllidaceae isoquinoline constituents and synthesis of related substances.a series of 23 amaryllidaceae isoquinoline alkaloids and related synthetic analogues were isolated or synthesized and subsequently evaluated in cell culture against the rna-containing flaviviruses (japanese encephalitis, yellow fever, and dengue viruses), bunyaviruses (punta toro, sandfly fever, and rift valley fever viruses), alphavirus (venezuelan equine encephalomyelitis virus), lentivirus (human immunodeficiency virus-type 1) and the dna-containing vaccinia virus. narciclasine [1], lycoricid ...19921336040
louping ill virus envelope protein expressed by recombinant baculovirus and vaccinia virus fails to stimulate protective immunity.we have constructed recombinant baculoviruses and vaccinia viruses containing cloned dna, encoding either the envelope protein alone or all of the structural proteins (core, membrane and envelope) of louping ill virus. glycosylated viral envelope protein, presented both inside and on the surface of insect and mammalian cells, was expressed by all four recombinant viruses. differences in antigenic presentation of the envelope protein were observed between the envelope protein and structural prote ...19921337231
augmentation of bovine leukemia virus (blv)-specific lymphocyte proliferation responses in ruminants by inoculation with blv env-recombinant vaccinia virus: their role in the suppression of blv replication.lymphocyte proliferation responses were investigated in sheep and cattle, in which the replication of bovine leukemia virus (blv) had been known to be suppressed by inoculation with recombinant vaccinia virus (rvv) expressing blv envelope glycoprotein (gp60). enhanced lymphocyte proliferation responses were observed in animals inoculated with rvv, regardless of whether they were naive or blv carriers. these responses were roughly inversely correlated to the growth of blv in the peripheral blood ...19921337572
[protective effect against blv by inoculation with a vaccinia virus-blv env recombinant]. 19921338242
vaccinia virus recombinants as potential herpes simplex virus vaccines. 19921338264
vaccinia and cowpox viruses encode a novel secreted interleukin-1-binding protein.supernatants from vaccinia virus (vv)-infected cv-1 cells were examined and found to contain a 33 kd protein capable of binding murine interleukin-1 beta (mil-1 beta). a vv open reading frame (orf) that exhibits 30% amino acid identity to the type ii il-1 receptor was expressed in cv-1-ebna cells and shown specifically to bind mil-1 beta. a similar orf from cowpox virus was expressed and also specifically bound mil-1 beta. a recombinant vv was constructed in which this orf was disrupted (vb15rko ...19921339315
proteins released from vaccinia cores are probably not involved in protein synthesis inhibition in vitro.1. it is known that vaccinia core proteins are released into the supernatant fraction when cores are incubated under appropriate conditions. when prepared in the absence of viral transcription this fraction inhibits in vitro protein synthesis. 2. we show here that after incubation, the cores loose the capability to inhibit protein synthesis. furthermore, we show that no inhibition of translation is observed with supernatant fractions prepared from transcribing cores. 3. sds-page analysis of the ...19921339506
[recurrent herpetic gingivostomatitis treated with moroxydine, antiherpin and the immunostimulant sre]. 19921340264
coevolution of hosts and microorganisms: an analysis of the involvement of cytokines in host-parasite interactions.parasites may employ particular strategies of eluding an immune response by taking advantage of those mechanisms that normally guarantee immunological self-tolerance. much in the way as it occurs during the establishment of self-tolerance, live pathogens may induce clonal deletion, functional inactivation (anergy) and immunosuppression. at this latter level, it appears that certain pathogens produce immunosuppressive cytokine-like mediators or provoke the host to secrete cytokines that will comp ...19921342705
the oct-1 pou domain mediates interactions between oct-1 and other pou proteins.the pou domain is the conserved dna binding domain of a family of gene regulatory proteins. it consists of a pou-specific domain and a pou homeodomain, connected by a variable linker region. oct-1 is a ubiquitously expressed pou domain transcription factor. it binds to the canonical octamer sequence (atgcaaat) as a monomer. here we show by chemical cross-linking and protein affinity chromatography that the oct-1 pou domain monomers can interact in solution. this association requires both the pou ...19921346336
preparation of candidate vaccinia-vectored vaccines for haemorrhagic fever with renal syndrome.two vaccinia-vectored candidate vaccines for haemorrhagic fever with renal syndrome were prepared by inserting cdna, representing the medium (m) genome segment, or the m and the small (s) genome segments of hantaan virus into the thymidine kinase gene of the connaught vaccine strain of vaccinia virus. in the single recombinant, the m segment was placed under control of the vaccinia virus 7.5 kda promoter. in the double recombinant, the m and s segments were placed under control of the vaccinia v ...19921347189
the age-dependent risk of postvaccination complications in vaccinees with smallpox vaccine.the use of vaccinia virus in recombinant systems may result in the occurrence of complications observed following smallpox vaccination. the results are presented of a large survey carried out in the ussr between 1968 and 1979. high complication rates are reported, particularly in older primary vaccinees; for example, there were 312.5 cases of neurological complications per million in those aged greater than or equal to 5 years. the author concludes that this factor will limit the use of vaccinia ...19921347190
immune response to a murine coronavirus: identification of a homing receptor-negative cd4+ t cell subset that responds to viral glycoproteins.the lymphocyte proliferative response to mouse hepatitis virus, strain jhm (mhv-jhm), a well-described cause of chronic and acute neurological infections, has been studied using vaccinia virus recombinants expressing individual mhv proteins. the surface (s) and transmembrane (m) glycoproteins were the most active proteins in causing proliferation of lymphocytes isolated from immunized adult mice, whereas lymphocytes from persistently infected mice proliferated only in response to the s protein. ...19921347668
the vaccinia virus k3l gene product potentiates translation by inhibiting double-stranded-rna-activated protein kinase and phosphorylation of the alpha subunit of eukaryotic initiation factor 2.interferon resistance of vaccinia virus is mediated by specific inhibition of phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eif-2 alpha) by the double-stranded-rna-activated (dai) protein kinase. vaccinia virus encodes a homolog of eif-2 alpha, k3l, the deletion of which renders the virus sensitive to interferon treatment. we have studied the mechanism by which this protein product elicits interferon resistance in a transient dna transfection system designed to evaluate ...19921347793
stimulation of lymphokines in jurkat cells persistently infected with vaccinia virus.the response of the human cd4+ t-cell line jurkat to infection with vaccinia virus was investigated. virus titers peaked approximately 3 to 4 days after infection, while cell growth paralleled that of uninfected cells, indicating that growth rates were not appreciably affected by viral infection. results from plaque assays and fluorescence-activated cell sorter (facs) analyses of virus antigens demonstrated that a persistent infection in which the percentage of infected cells and the virus titer ...19921347794
specific lysis of targets expressing varicella-zoster virus gpi or gpiv by cd4+ human t-cell clones.varicella-zoster virus (vzv)-specific cd4-positive t cells are known to lyse targets expressing vzv antigen, but little is known of the glycoprotein specificity or phenotype of these cells. to test the ability of t cells to distinguish between gpi and gpiv (which share an antibody-defined epitope), we prepared clones from blood from four healthy individuals by limiting dilution. among 68 t-cell clones from four donors which were vzv specific in tests of proliferation, 30 lysed autologous epstein ...19921348545
analysis of murine major histocompatibility complex class ii-restricted t-cell responses to the flavivirus kunjin by using vaccinia virus expression.the present paper analyzes the influence of major histocompatibility complex (mhc) class ii (ir) genes on mhc class ii-restricted t-cell responses to west nile virus (wnv) and recombinant vaccinia virus-derived kunjin virus antigens and identifies the immunodominant kunjin virus antigens. generally, mice were primed by intravenous infection with wnv or kunjin virus, and their cd4+ t cells were stimulated in vitro 14 days later with wnv or kunjin virus antigens to pulse macrophage or b-cell antig ...19921349926
the e3l gene of vaccinia virus encodes an inhibitor of the interferon-induced, double-stranded rna-dependent protein kinase.a vaccinia virus-encoded double-stranded rna-binding protein, p25, has been previously implicated in inhibition of the interferon-induced, double-stranded rna-activated protein kinase. in this study, we have identified the vaccinia viral gene (wr strain) that encodes p25. amino acid sequence analysis of a chymotryptic fragment of p25 revealed a close match to the vaccinia virus (copenhagen strain) e3l gene. the wr strain e3l gene was cloned and expressed either in cos-1 cells or in rabbit reticu ...19921350676
immunisation with canarypox virus expressing rabies glycoprotein.poxviruses have many useful features as vectors for genes that carry immunising antigens from other viruses, such as ease of production and induction of cellular and humoral immunity, but there is concern about the safety of vaccinia virus. we turned to an avian poxvirus (canarypox); this virus undergoes abortive replication in mammalian cells that enables presentation of early gene products to the immune system. canarypox virus was used as a vector for the rabies glycoprotein g gene. the safety ...19921351126
dna-binding activity of the murine homeodomain protein hox-2.3 produced by a hybrid phage t7/vaccinia virus system.homeobox-containing genes encode transcription factors that, via the homeodomain, bind specifically to dna. to study the dna-binding properties of the murine homeodomain-containing protein, hox-2.3, a hybrid expression system was used, combining gene expression by recombinant vaccinia virus (revv) with bacteriophage t7 transcription. expression was achieved by co-infecting hela cells with two revvs, one expressing the t7-rna polymerase-encoding gene directed by the vv promoter, p7.5, and another ...19921353046
vaccination of vaccinia-naive adults with human immunodeficiency virus type 1 gp160 recombinant vaccinia virus in a blinded, controlled, randomized clinical trial. the aids vaccine clinical trials network.the safety and immunogenicity of a human immunodeficiency virus type 1 (hiv-1) gp160 recombinant vaccinia virus (hivac-1e) vaccine was evaluated in vaccinia-naive, healthy adults at low risk for acquiring hiv-1 infection. volunteers (n = 36) were randomized to receive hivac-1e or control vaccinia virus at two dosages by bifurcated needle puncture at 0 and 2 months; 12 hivac-1e and 6 control vaccinia virus recipients received either 10(6) or 10(7) pfu/ml at each inoculation. there was no signific ...19921353102
transition from rapid processive to slow nonprocessive polyadenylation by vaccinia virus poly(a) polymerase catalytic subunit is regulated by the net length of the poly(a) tail.the mrna of vaccinia virus, like that of eukaryotes, possesses a poly(a) tail. vp55, the catalytic subunit of the heterodimeric vaccinia virus poly(a) polymerase, was overexpressed and purified to near homogeneity. vp55 polyadenylated a 30-mer primer representing the 3' end of a vaccinia virus mrna bimodally: 30-35 adenylates were added in a rapid, processive, initial burst, after which polyadenylation decelerated dramatically and became nonprocessive. polyadenylation of variants of the 30-mer p ...19921353739
expression of cytokines by recombinant vaccinia viruses: a model for studying cytokines in virus infections in vivo. 19921354650
on the antiviral activity of diffusomycin (oxazolomycin).the effect of the beta-lactone antibiotic diffusomycin (oxazolomycin) was investigated against vaccinia (lister), herpes simplex type 1 (kupka), influenza a (wsn; h1n1), and coxsackie a9 viruses. diffusomycin reduced significantly the plaque formation of enveloped dna and rna viruses by more than 90% in the range of the maximally tolerated dose. as could be shown with vaccinia virus, the antiviral action was not caused by virucidal effect on virions or by interaction with virus adsorption and pe ...19921359768
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