the gtpase rab3a negatively controls calcium-dependent exocytosis in neuroendocrine cells.there is accumulating evidence that small gtpases of the rab family regulate intracellular vesicle traffic along biosynthetic and endocytotic pathways in eukaryotic cells. it has been suggested that rab3a, which is associated with synaptic vesicles in neurons and with secretory granules in adrenal chromaffin cells, might regulate exocytosis. we report here that overexpression in pc12 cells of rab3a mutant proteins defective in either gtp hydrolysis or in guanine nucleotide binding inhibited exoc ...19948187757
peptide immunogen mimicry of putative e1 glycoprotein-specific epitopes in hepatitis c virus.hepatitis c virus (hcv) accounts for most cases of acute and chronic non-a and non-b hepatitis with serious consequences that may lead to hepatocellular carcinoma. the putative envelope glycoproteins (e1 and e2) of hcv probably play a role in the pathophysiology of the virus. in order to map the immunodominant domains of the e1 glycoprotein, two epitopes from amino acid residues 210 to 223 (p1) and 315 to 327 (p2) were predicted from the hcv sequence. immunization of mice with the synthetic pept ...19948207814
immunological characterization of the feline herpesvirus-1 glycoprotein b and analysis of its deduced amino acid sequence.feline herpesvirus 1 (fhv-1) is an important viral pathogen of cats. like other alphaherpesviruses, fhv-1 contains a hsv-1 glycoprotein b (gb) homolog. in this study, monospecific antisera to hsv-1 gb reacted with three fhv-1 proteins (100, 64, and 58 kda) present in virion lysates by immunoprecipitation and immunoblot analyses. reduced stringency hybridization experiments using a hsv-1 gb probe localized the fhv-1 gb gene to a 9.6-kb sa/l fragment in the unique long region of the genome. northe ...19938212548
identification of two human rho gdp dissociation inhibitor proteins whose overexpression leads to disruption of the actin cytoskeleton.proteins (ief's 1120, 8118, 8120) sharing similarity to the bovine rho gdp dissociation inhibitor (gdi) have been identified in the human two-dimensional-gel database of keratinocyte proteins. molecular cloning of the corresponding cdnas showed that ief 8118 is the human homolog of bovine gdi while ief 8120 is a distinct although related protein. all available information indicates the ief 1120 is a derivative of ief 8120. the cdnas coding for ief's 8118 and 8120 were recombined into vaccinia vi ...19938262133
recombinant viral vaccines for enzootic bovine leucosis.recently published studies on the development and use of recombinant vaccinia virus (vv) vaccines incorporating either the complete envelope (env) gene or only a fragment of the env gene consisting of the coding sequence for the env glycoprotein 51 (gp51) and part of gp30 of the bovine leukaemia virus (blv) are described. it has been reported that vaccination of sheep with recombinant vv vaccines containing the complete env gene appears to protect sheep against challenge infection with blv. the ...19938270269
an immunodominant cytotoxic t cell epitope on the vp7 rotavirus protein overlaps the h2 signal peptide.c57bl/6 (h-2b) mice were primed with the bovine rf strain of rotavirus to study the induction of cd8+ cytotoxic t lymphocytes (ctls). these rotavirus-specific ctls were detected only after in vitro restimulation with the virus. using a recombinant vaccinia virus we identified the rf vp7 protein as a major target of these ctls. the response against this protein was obtained also after in vitro restimulation with simian sa11 and human wa strains of rotavirus. using published db and kb allele-speci ...19938277264
single capripoxvirus recombinant vaccine for the protection of cattle against rinderpest and lumpy skin disease.a recombinant capripoxvirus has been constructed containing a full-length cdna of the fusion protein gene of rinderpest virus. the gene was inserted in the thymidine kinase gene of the capripox genome under the control of the vaccinia virus major late promoter p11 together with the escherichia coli gpt gene in the opposite orientation under the control of the vaccinia early/late promoter p7.5. a vaccine prepared from this recombinant virus protected cattle against clinical rinderpest after a let ...19938342321
role of n-linked glycans in antigenicity, processing, and cell surface expression of bovine herpesvirus 1 glycoprotein giv.glycoprotein giv, a structural component of bovine herpesvirus type 1, stimulates high titers of virus-neutralizing antibody. the protein contains three potential sites for the addition of n-linked carbohydrates. three mutants were constructed by oligonucleotide-directed mutagenesis, in each case changing one n-linked glycosylation site from asn-x-thr/ser to ser-x-thr/ser. a fourth mutant was altered at two sites. the altered forms of the giv gene were cloned into a vaccinia virus transfer vecto ...19938380463
protection of cattle from bhv-1 infection by immunization with recombinant glycoprotein giv.high levels of recombinant bovine herpesvirus-1 (bhv-1) glycoprotein iv were produced in baculovirus, adenovirus, vaccinia virus and escherichia coli expression systems. the different recombinant forms as well as authentic giv were injected intramuscularly into seronegative calves. with the exception of e. coli-produced giv, all forms of giv induced high levels of neutralizing antibodies both in the serum and in the nasal superficial mucosa. animals immunized with giv produced in insect or mamma ...19938381248
identification of different target glycoproteins for bovine herpes virus type 1-specific cytotoxic t lymphocytes depending on the method of in vitro stimulation.vaccinia virus recombinants expressing the three major bovine herpes virus-1 (bhv-1) glycoproteins gi, giii and giv were used to identify the major target antigens for bhv-1-specific ctl isolated from immune cattle. peripheral blood mononuclear cells (pbmc) expanded in vitro in the presence of interleukin-2 (il-2) and lysed both giii- and giv-infected target cells. secondary in vitro stimulation of pbmc was also performed in the presence of either fixed bhv-1-infected autologous fibroblasts or u ...19938382189
analysis of bovine herpesvirus 1 glycoprotein giv truncations and deletions expressed by recombinant vaccinia viruses.glycoprotein giv is an envelope component of bovine herpesvirus type 1 and appears to be involved in attachment, penetration, and cell fusion. four antigenic domains which include both continuous and discontinuous epitopes have been previously defined by competition binding assays using giv-specific monoclonal antibodies (mabs). here we describe the construction of c-terminal truncations and internal deletions in the giv-encoding gene and analyses of the effects of these mutations on the synthes ...19938383232
synthesis and assembly of infectious bovine papillomavirus particles in vitro.bovine papillomavirus type 1 (bpv-1) virions were produced in vitro using vaccinia virus (vv) recombinants expressing the bpv-1 l1 and l2 capsid proteins. particles morphologically resembling papillomaviruses were observed in the nucleus of cells infected with a vv recombinant for the bpv-1 l1 protein, and greater numbers of similar particles were seen in the nuclei of cells infected with a vv double recombinant for l1 and l2. virus-like particles (vlps) assembled in cells infected with the vv d ...19938385700
processing of the envelope glycoproteins of pestiviruses.the genomic rna of pestiviruses is translated into a large polyprotein that is cleaved into a number of proteins. the structural proteins are n terminal in this polyprotein and include three glycoproteins called e0, e1, and e2 on the basis of the order in which they appear in the polyprotein. using pulse-chase experiments, we show that a pestiviral glycoprotein precursor, e012, is formed that is processed into e0, e1, and e2 in an ordered fashion. processing is initiated by a nascent cleavage be ...19938388499
immunological responses of mice and cattle to baculovirus-expressed f and h proteins of rinderpest virus: lack of protection in the presence of neutralizing antibody.rinderpest is a highly contagious viral disease of ruminants and has greater than 95% morbidity and mortality. the etiological agent, rinderpest virus (rpv), is a member of the family paramyxoviridae and the genus morbillivirus. immune responses to both the hemagglutinin (h) and the fusion (f) antigens of morbilliviruses play an important role in the prevention of infection, and only attenuated live vaccines have been shown to provide protective immunity against the group. the lack of protection ...19938437215
immunisation of cattle with a recombinant vaccinia vector expressing the haemagglutinin gene of rinderpest virus.the efficacy of a recombinant rinderpest vaccine, constructed by inserting the rinderpest virus haemagglutinin gene into attenuated vaccinia virus, lc16mo strain, was tested in cattle. after subcutaneous inoculation of 10(8) plaque-forming units (pfu) of the recombinant vaccine, neither palpable skin lesions nor increases in body temperature were observed, indicating the absence of detectable clinical reactions. all the vaccinated cattle were completely protected from challenge with the saudi 1/ ...19938456545
involvement of cd8+ t cells in delayed-type hypersensitivity responses against bovine leukemia virus (blv) induced in sheep vaccinated with recombinant vaccinia virus expressing blv envelope glycoprotein.delayed-type hypersensitivity responses against bovine leukemia virus (blv) envelope glycoprotein (gp60) were induced in the skin of sheep vaccinated with recombinant vaccinia virus (rvv) expressing blv glycoprotein. the lesions were characterized by marked infiltration of lymphocytes, slight migration of neutrophils, eosinophils, and macrophages in the dermis to hypodermis, and partial intercellular edema in the reticular layer. immunohistochemical analysis with monoclonal antibodies demonstrat ...19938470333
lymphocyte proliferative responses to recombinant bovine herpes virus type 1 (bhv-1) glycoprotein gd (giv) in immune cattle: identification of a t cell epitope.the lymphocyte proliferative response to bhv-1 in immune cattle was compared to recombinant wild-type gd and truncated gd produced from recombinant vaccinia viruses. the response exhibited by recombinant proteins was comparable to the response induced by bhv-1 suggesting that gd is the major target structure for stimulation of bovine lymphocytes. analysis of the proliferative response using vaccinia virus vectors expressing various modified forms of gd identified a region between residues 165 an ...19958546801
vaccinia virion protein i8r has both dna and rna helicase activities: implications for vaccinia virus transcription.a nucleic acid-dependent atpase was purified from vaccinia virions and shown to have both dna:dna and rna:rna helicase activities. this is only the third helicase to be identified that can unwind both dna and rna duplexes. the dna helicase activity copurified with nucleoside triphosphate phosphohydrolase ii (nphii), an rna helicase encoded by gene i8r (s. shuman, proc. natl. acad. sci. usa 89:10935-10939, 1992). immunodepletion with two antisera to nphii and analysis of recombinant nphii protein ...19968551617
identification and characterization of a bovine herpesvirus-1 (bhv-1) glycoprotein gl which is required for proper antigenicity, processing, and transport of bhv-1 glycoprotein gh.dna sequence analysis of the bovine herpesvirus-1 (bhv-1) genome revealed the presence of an open reading frame named ul1 which exhibited limited homology to glycoprotein gl of herpes simplex virus-1 (s. k. khattar, s. van drunen littel-van den hurk, l. a. babiuk, and s. k. tikoo, virology 213, 28-37). to identify the bhv-1 ul1 protein, rabbit antisera were prepared against two synthetic peptides that were predicted by computer analysis to encompass antigenic epitopes. sera against both peptides ...19968623555
vaccinia virus-expressed bovine ephemeral fever virus g but not g(ns) glycoprotein induces neutralizing antibodies and protects against experimental infection.two related glycoproteins (g and g(ns)) encoded in the bovine ephemeral fever virus (befv) genome were expressed from recombinant vaccinia viruses (rvv). both proteins were detected in lysates of rvv-infected cells by labelling with d-[6-3h]glucosamine or by immuno-blotting. the recombinant g protein (mol. mass 79 kda) appeared slightly smaller than the native g protein but reacted with monoclonal antibodies directed against all defined neutralizing antigenic sites (g1, g2, g3a, g3b and g4). the ...19968627251
bovine herpesvirus 1 u(s) open reading frame 4 encodes a glycoproteoglycan.sequence analysis of the short unique (us) segment of the bovine herpesvirus 1 (bhv-1) genome predicted that the us open reading frame (orf) 4 encodes a protein with homology to glycoprotein g (gg) of other alpha-herpesviruses (p. leung-tack, j.-c. audonnet, and m. riviere, virology 199:409-421, 1994). rna analysis showed that the us orf4 is contained within two transcripts of 3.5 and 1.8 kb. the 3.5 kb rna represents a structurally bicistronic rna which encompasses the us orf3 and us orf4, wher ...19968627780
new neplanocin analogues. 6. synthesis and potent antiviral activity of 6'-homoneplanocin a1.the design, synthesis, and antiviral activities of 6'-homoneplanocin a (hnpa, 3) and its congeners having nucleobases other than adenine, such as 3-deazaadenine (4), guanine (5), thymine (6), and cytosine (7), were described. starting from the known cyclopentenone derivative 8, the optically active (mesyloxy)cyclopentene derivative 15 was prepared, which was condensed with nucleobases then deprotected to give target compounds 3-7. of these compounds, hnpa showed an antiviral activity spectrum th ...19968691433
recombinant decorin glycoforms. purification and structure.the vaccinia virus/t7 bacteriophage expression system was used to express human decorin in ht-1080 cells by co-infection with vtf7-3, encoding t7 rna polymerase, and vdcn1, encoding the decorin core protein fused to a polyhistidine-insulin signal sequence fusion-protein cassette. overexpression using the vaccinia virus/t7 phage system resulted in secretion of approximately 30 mg of decorin/10(9) cells per 24 h which enabled purification and separation of multiple glycoforms under native conditio ...19968702652
the bovine herpesvirus type 1 major tegument protein vp8 expressed in recombinant vaccinia virus does not induce significant immunity in mice.we previously reported the characterization of the gene encoding the bovine herpesvirus type 1 (bhv-1) major tegument protein vp8. with the aim of defining the immunological properties of this protein, we constructed a recombinant vaccinia virus (vv-vp8) in which expression of the vp8 gene was regulated by the p7.5 early/late promoter. since the sequence of the vp8 gene contained a tttttnt motif known to serve as a transcription termination signal of vaccinia virus genes of the early class, a se ...19968725115
viral status and antibody response in cattle inoculated with recombinant bovine leukemia virus-vaccinia virus vaccines after challenge exposure with bovine leukemia virus-infected determine the bovine leukemia virus (blv) antibody response and infectivity status in blv-vaccinated cattle after challenge exposure with blv-infected lymphocytes.19968725805
enzyme-linked immunosorbent assay using recombinant antigens for serodiagnosis of japanese encephalitis.recombinant japanese encephalitis (je) virus proteins were evaluated as antigens for serodiagnosis of je using an enzyme-linked immunosorbent assay (elisa). the premembrane/membrane (prm/m) and envelope (e) proteins of je virus were expressed in hela cells infected with a recombinant vaccinia virus that encodes the je virus prm and e genes and were released from cells in a particulate form. the particulate antigens were partially purified from culture fluid from the infected cells by precipitati ...19968825714
genomic analysis of a transposition-deletion variant of orf virus reveals a 3.3 kbp region of non-essential dna.restriction endonuclease analysis of the dna extracted orf virus strain nz2, which had been serially passaged in primary bovine testis cells, revealed a population of variants that had over-grown the wild-type virus. at least three distinct mutant forms were identified in which the right end of the genome had been duplicated and translocated to the left end, accompanied by deletions of sequences at the left end. sequencing of a single variant isolated from the heterogeneous population revealed t ...19958847502
progress on vaccines against parasites.there has been significant progress in attempts to develop effective vaccines against parasitic diseases, including malaria, leishmaniasis and schistosomiasis. in malaria, in addition to field trials with spf66, the colombian malaria vaccine, several plasmodium falciparum candidate vaccines are under phase i testing, including nyvac-7, a multi-antigen, attenuated recombinant vaccinia virus. additional candidate antigens are at an advanced stage of pre-clinical development. in leishmaniasis, phas ...19968854004
cytopathogenicity of a pestivirus correlates with a 27-nucleotide insertion.cytopathogenic (cp) bovine viral diarrhea virus (bvdv) strains are generated in cattle persistently infected with noncytopathogenic (noncp) bvdv.cp bvdv strains are considered crucial for the development of fatal mucosal disease. comparative analysis of cp and noncp bvdv strains isolated from one animal suffering from mucosal disease revealed that the genomes of the cp bvdv strain (cp7) and the corresponding noncp bvdv strain (ncp7) are highly homologous. however, only the genome of cp7 contains ...19968892907
identification and characterization of the bovine herpesvirus 5 us4 gene and gene products.the bhv-5 strain n569 (bhv-5/n569) homolog to the bhv-1 us4 gene was sequenced and characterized. rna analyses showed that a 1.8-kb mrna which contains the bhv-5/n569 us4 open reading frame initiates 55 nucleotides upstream from the predicted translational start codon and terminates 17 nucleotides downstream from the consensus sequence for polyadenylation. comparison of the deduced amino acid sequences of the predicted us4 encoded proteins of bhv-5/n569 and bhv-1 strain schönböken (bhv-1/schö) r ...19968918539
novel viral vaccines for livestock.recent advances in our understanding of virulence factors of viruses and the proteins or glycoproteins involved in inducing neutralizing antibodies or cell mediated immunity are forming the foundation for the development of a new generation of viral vaccines. using bovine herpesvirus as an example, we have identified glycoproteins gb, gc, and gd as important targets for inducing neutralizing antibody responses, with gd being able to induce the highest neutralizing and cellular responses. for sub ...19968988880
rescue of rinderpest virus from cloned cdna.rinderpest virus is a morbillivirus and is the causative agent of a widespread and important disease of cattle. the viral genome is a single strand of rna in the negative sense. we have constructed plasmids containing cdna copies of the 5' and 3' termini of the virus separated by a reporter gene and have shown that antigenome-sense rna transcripts of these model genomes can be replicated, transcribed, and packaged by helper virus, both rinderpest virus and the related measles virus. further, the ...19978995650
[jenner's cowpox vaccine in light of current vaccinology].two hundred years ago edward jenner inoculated james phipps with vaccinia and 181 years later smallpox had disappeared from the surface of the earth as a result of generalized vaccination. compared to the requirements of modern vaccinology, the procedures used by jenner and his successors, were extremely primitive because of an almost total lack of knowledge in the field of microbiology and immunology. the active principle of smallpox vaccine is vaccinia virus, which in many respects, differs fr ...19969027132
protective immunity in cattle following vaccination with conventional and marker bovine herpesvirus-1 (bhv1) vaccines.the efficacy of two experimental subunit gd vaccines and two commercial whole virus vaccines was determined in a bovine herpesvirus-1 (bhv1) challenge trial. full-length gd and a truncated, secreted form of gd (tgd) were produced using a vaccinia virus expression system and purified by affinity chromatography. comparison of these forms of gd did not reveal significant structural or antigenic differences. calves immunized with gd or tgd in avridine developed significantly (p < 0.05) higher neutra ...19979041664
a novel strategy for determining protective antigens of the parapoxvirus, orf virus.we investigated the feasibility of using vaccinia virus (vac) recombinants containing large multigene fragments of orf virus dna to identify protective antigens of orf virus (ov). sixteen ov strain nz2 dna fragments with an average size of 11.4 kb were recombined into vac strain lister. each fragment was mapped relative to ov restriction endonuclease maps but was otherwise uncharacterized. together the recombinants represent 95% of the ov genome in an overlapping manner. immunofluorescence showe ...19979123861
chlamydia psittaci inca is phosphorylated by the host cell and is exposed on the cytoplasmic face of the developing inclusion.chlamydiae are obligate intracellular bacteria that replicate within a non-acidified vacuole called an inclusion. chlamydia psittaci (strain gpic) produces a 39 kda protein (inca) that is localized to the inclusion membrane. while inca is present as a single 39 kda species in purified reticulate bodies, two additional higher m(r) forms are found in c. psittaci-infected cells. this finding suggested that inca may be post-translationally modified in the host cell. here we present evidence that inc ...19979140978
mapping of the target antigens of the rotavirus-specific cytotoxic t cell response.the cytotoxic t cell (ctl) response in c57/b16 (h-2b) mice to rotavirus has been analysed using a cognate set of vaccinia virus recombinants covering the 12 primary gene products of the uktc strain of bovine rotavirus. the gene products of rna segments 5 (vp5/nsp-1) and 8 (vp7) both elicited a classic cd8+ class i mhc restricted ctl response. using l cells transfected with specific class i mhc loci as targets the vp5/nsp-1 response was found to be restricted at db and the vp7 response at kb. vac ...19979152424
serine protease of pestiviruses: determination of cleavage sites.the single-stranded genomic rna of pestiviruses is of positive polarity and encompasses one large open reading frame of about 4,000 codons. the resulting polyprotein is processed co- and posttranslationally by virus-encoded and host cell proteases to give rise to the mature viral proteins. a serine protease residing in the nonstructural (ns) protein ns3 (p80) has been shown to be essential for the release of the ns proteins located downstream of ns3. in this report the ns3 serine protease-depend ...19979188613
analysis of bovine respiratory syncytial virus envelope glycoproteins in cell compare the requirements for respiratory syncytial virus (rsv)-mediated cell fusion, the fusion (f), attachment (g) and small hydrophobic (sh) glycoproteins of bovine rsv (brsv), ovine rsv (orsv) and human rsv (hrsv) were expressed individually or coexpressed in either homologous or heterologous combinations in hela cells, using the vaccinia virus-t7 polymerase transient expression system. cell fusion was examined by a reporter gene activation assay. although the expression of the f protein a ...19979266983
antiviral activity of an extract from leaves of the tropical plant acanthospermum hispidum.incubation of the alphaherpesviruses pseudorabiesvirus (prv) and bovine herpesvirus 1 during infection of cell cultures with an extract prepared from the leaves of acanthospermum hispidum impaired productive replication of these viruses in a concentration-dependent manner whereas propagation of classical swine fever virus, foot-and-mouth disease virus and vaccinia virus was not affected. the 50% inhibitory concentration for cell growth (ic50) was 107 +/- 5 microliters/ml, and the concentration r ...19979330761
role of envelope glycoproteins of bovine respiratory syncytial virus in cell investigate the requirements for bovine respiratory syncytial virus (brsv) cell fusion, the fusion (f), the attachment (g) and the small hydrophobic (sh) glycoproteins were expressed individually or coexpressed, using the vaccinia virus-t7 polymerase transient expression system. the contribution of individual glycoproteins in cell fusion was studied by a reporter gene activation assay. activation of a reporter gene, beta-galactosidase, was assessed by colorimetric assay of detergent cell lysa ...19979343948
recombinant vaccinia viruses expressing the f, g or n, but not the m2, protein of bovine respiratory syncytial virus (brsv) induce resistance to brsv challenge in the calf and protect against the development of pneumonic lesions.the immunogenicity and protective efficacy of recombinant vaccinia viruses (rvv) encoding the f, g, n or m2 (22k) proteins of bovine respiratory syncytial virus (brsv) were evaluated in calves, the natural host for brsv. calves were vaccinated either by scarification or intratracheally with rvv and challenged 6 to 7 weeks later with brsv. although replication of rvv expressing the f protein in the respiratory tract was limited after intratracheal vaccination, the levels of serum and pulmonary an ...19979400970
molecular genetic analyses of parapoxviruses pathogenic for humans.the current members of the genus parapoxvirus are orf virus (orfv), bovine papular stomatitis virus (bpsv), pseudocowpoxvirus (pcpv) and parapoxvirus of red deer in new zealand (pvnz). bpsv and pcpv are maintained in cattle while orfv is maintained in sheep and goats, but all three are zoonoses. only the recently reported pvnz has yet to be recorded as infecting humans. tentative members of the genus are camel contagious ecthyma virus, chamois contagious ecthyma virus and sealpoxvirus. the separ ...19979413523
demonstration of the specificity of poliovirus encapsidation using a novel replicon which encodes enzymatically active firefly luciferase.the specificity of poliovirus encapsidation has been studied using a novel chimeric genome in which the gene encoding firefly luciferase has been substituted for the vp2-vp3-vp1 genes of the poliovirus capsid (p1) gene. transfection of rna transcribed in vitro from this genome resulted in a vp4-luciferase fusion protein which retained luciferase enzyme activity. since the detection of enzyme activity was dependent upon replication of the transfected rna genome, we refer to these genomes as repli ...19989527910
recombinant swinepox virus expressing beta-galactosidase: investigation of viral host range and gene expression levels in cell culture.swinepox virus (spv) has been proposed as a potential vector for generating recombinant vaccines for swine. however, little is known about important aspects of spv biology, such as the functionality of spv promoters or the host range of spv. using a transient expression assay, well-characterized vaccinia virus promoters were shown to be active in cells infected with spv. a recombinant spv expressing beta-galactosidase (beta-gal) was constructed and characterized. the e. coli lacz gene was placed ...19989568038
detection, cdna cloning and sequencing of canine interleukin canine peripheral blood mononuclear cells (pbmc) the mrnas coding for both subunits of canine interleukin 12 (il-12) were identified using reverse transcription polymerase chain reaction (rt-pcr). stimulation of canine pbmc with staphylococcus aureus strain cowan plus concanavalin a for 5 h resulted in significant mrna synthesis. likewise, inactivated vaccinia virus induced il-12 mrna synthesis, however with different kinetics. the complete nucleotide sequence for both il-12 subunits was dete ...19989617567
evidence for incomplete replication of a penguin poxvirus in cells of mammalian origin.the recent discovery of a novel poxvirus [penguin-pox virus (ppv)] from jackass penguins offers the potential of a unique candidate vaccine vector for use in mammals. infectivity studies were therefore undertaken using a number of mammalian cell lines and chick embryo fibroblasts (cef). it was shown that the simian cv-1 cell line was able to support replication of the ppv dna, but no infectious progeny virus could be recovered from the infected cells. electron microscopy was used to establish th ...19989680125
vaccinia virus protein b18r inhibits the activity and cellular binding of the novel type interferon-delta.the soluble vaccinia virus-encoded protein b18r inhibits the antiviral activity and cellular binding of the type i interferons (ifn)-alpha, -beta and -omega of different mammalian species. recently, a novel type i ifn was detected in pigs and classified as a member of a distinct ifn family designated ifn-delta. our study aimed to determine if the structural properties of this shortest (149 residues long) type i ifn allow its interaction with the type i ifn-binding protein b18r. experiments using ...19989680126
delivery of the theileria parva p67 antigen to cattle using recombinant vaccinia virus: il-2 enhances evaluate vaccinia virus as a delivery system for recombinant antigen in cattle, calves were immunized with a recombinant vaccinia virus (rvv) expressing the sporozoite surface antigen (p67) of theileria parva (v-67) combined with those expressing bovine il-4 (v-il4) or il-2 (v-il2). the anti-p67 antibody levels detected in calves inoculated with the combination of v-67 and v-il4 were higher than those produced by animals injected with v-67 alone or v-67 and v-il2. on challenge with cryopreser ...19989682391
vaccinia virus-encoded cytokine receptor binds and neutralizes chicken counteract the host immune response, poxviruses have evolved secreted factors that bind cytokines and thereby neutralize their biological activities. the vaccinia virus b8r gene encodes a protein that neutralizes interferon-gamma (ifn-gamma) from several mammals including man, cow, rat, and rabbit but not mice. we now report that the activity of the b8r gene product is not restricted to cytokines of mammals: it also efficiently neutralized chicken ifn-gamma. b8r blocked chicken ifn-gamma-medi ...19989721232
sequence analysis of a functional polymerase (l) gene of bovine respiratory syncytial virus: determination of minimal trans-acting requirements for rna replication.the complete nucleotide sequence of a functional clone of the large polymerase (l) gene of bovine respiratory syncytial virus (brsv) strain a51908 was determined by analysis of cloned cdnas obtained from genomic and mrnas. the brsv l gene is 6573 nt in length and the derived polypeptide has 2162 aa. alignment of the sequences of the brsv l gene, and its encoded protein, with sequences of the l gene and protein of human respiratory syncytial virus strain a2 showed 77% identity at the nucleotide l ...19989747733
generation of bovine respiratory syncytial virus (brsv) from cdna: brsv ns2 is not essential for virus replication in tissue culture, and the human rsv leader region acts as a functional brsv genome order to generate recombinant bovine respiratory syncytial virus (brsv), the genome of brsv strain a51908, variant atue51908, was cloned as cdna. we provide here the sequence of the brsv genome ends and of the entire l gene. this completes the sequence of the brsv genome, which comprises a total of 15,140 nucleotides. to establish a vaccinia virus-free recovery system, a bhk-derived cell line stably expressing t7 rna polymerase was generated (bsr t7/5). recombinant brsv was reproducibly recov ...19999847328
the reactivity of monoclonal antibodies against orf virus with other parapoxviruses and the identification of a 39 kda immunodominant protein.a panel of 27 mouse monoclonal antibodies (mabs) was raised against orf virus. sixteen of these mabs reacted with a protein with a molecular mass of 65 kda, 8 reacted with a protein with a molecular mass of 39 kda and three remain uncharacterised. reactivity of the mabs with a library of recombinant vaccinia viruses expressing various regions of the nz-2 orf virus genome identified the approximate positions of the genes encoding these 2 immunodominant orf virus proteins. the gene encoding the 39 ...19989930187
development and in vitro characterization of recombinant vaccinia viruses expressing bovine leukemia virus gp51 in combination with bovine il4 or il12.type 1 and type 2 immune responses are modulated by il12 or il4, respectively, at the time of lymphocyte priming. importantly, type 1 responses have been associated with resistance to retroviral infection in mice, humans, and ruminants. specifically, vaccination of sheep with vaccinia virus expressing bovine leukemia virus (blv) gp51 resulted in protective immunity with the characteristics of a type 1 response, whereas vaccination of cattle resulted in a non-protective type 2 response. in order ...19999987178
mammalian cell expression of malaria merozoite surface proteins and experimental dna and rna immunisation.the gene for a 45 kda merozoite surface protein (msa-2) of the human malaria parasite plasmodium falciparum was pcr amplified and cloned into eukaryotic expression vectors vr1012 and pcdna3 to yield plasmids p1 and p2, respectively. the coding sequences for two n-terminal fragments of the 185 kda merozoite surface protein (msa-1) gene were similarly pcr amplified and cloned into vectors vr1020 and vr1012 to yield plasmids p3 and p4, respectively. the msa-1 signal peptide sequence, present in p4, ...19999989240
expression of the e2 envelope glycoprotein of bovine viral diarrhoea virus (bvdv) elicits virus-type specific neutralising antibodies.a region of genome from the nadl strain of bvdv corresponding to the coding sequence for the e2 glycoprotein has been molecularly cloned using rt-pcr. the viral cdna sequence was used to construct vaccinia virus recombinants that expressed either the entire e2 coding sequence or fragments of it. these recombinants were used to immunise mice of three h-2 haplotypes to investigate their ability to elicit a neutralising antibody response against bvdv. sera from mice immunised with the recombinant e ...199910078593
virus reduction in the preparation of intravenous immune globulin: in vitro experiments.while immune globulins for intravenous administration (igiv) have an excellent record with respect to virus safety, concern regarding these preparations has been raised by reports of transmission of hepatitis c virus (hcv) to patients treated with igiv and the presence of genetic material for hcv in igiv preparations.199910204586
the hemagglutinin-esterase of mouse hepatitis virus strain s is a comparative analysis of the hemagglutinin-esterase (he) protein of mouse hepatitis virus strain s (mhv-s) and the he protein of influenza c virus, we found major differences in substrate specificities. in striking contrast to the influenza c virus enzyme, the mhv-s esterase was unable to release acetate from bovine submandibulary gland mucin. furthermore, mhv-s could not remove influenza c virus receptors from erythrocytes. analysis with free sialic acid derivatives revealed that the mhv-s he ...199910233932
rotavirus-specific cytotoxic t lymphocytes recognize overlapping epitopes in the amino-terminal region of the vp7 glycoprotein.rotavirus-specific cytotoxic t lymphocytes (ctl) play an important role in the resolution of rotavirus infection. the outer capsid glycoprotein, vp7, elicits a class i mhc-restricted ctl response. vaccinia virus recombinants expressing the vp7 genes from simian rotavirus sa11 (serotype g3) and from the rf strain of bovine rotavirus (serotype g6) were used to analyze the ctl activity to this antigen in balb/c (h-2(d)) and c57bl/6 (h-2(b)) mice neonatally infected with homologous and heterologous ...199910329553
cell-mediated immune responses in cattle vaccinated with a vaccinia virus recombinant expressing the nucleocapsid protein of rinderpest virus.rinderpest virus (rpv) is a member of the genus morbillivirus in the family paramyxoviridae which causes an acute and often fatal disease in large ruminants. to examine the immune response to the virus nucleocapsid (n) protein, a recombinant vaccinia virus expressing rpv nucleocapsid protein (rvv-rpv-n) was used to vaccinate cattle. the recombinant vaccine induced low levels of non-neutralizing anti-n antibodies. rpv-specific cell-mediated immunity induced by the recombinant was assessed by meas ...199910423130
antibodies produced by mice immunized with recombinant vaccinia viruses expressing two different types of a major theileria sergenti surface antigen (p32) react with the native surface antigen.a 32 kda major surface antigen, p32, of theileria sergenti at the piroplasm stage is the main target of the host immune response. the immunogenic property of the p32 varies in some strains among the population of theileria sergenti in japan where the chitose type and the ikeda type are the most common varieties. we have constructed vaccinia virus recombinants vv/p32c and vv/p32i which harbor the chitose and ikeda types of p32 gene, respectively. it was found that vv/p32c and vv/p32i produced typ ...199910435791
identification and characterisation of an early/late bi-directional promoter of the capripoxvirus, lumpy skin disease virus.identification and characterisation of an early/late bi-directional promoter element of lumpy skin disease virus (lsdv) is described. the 56 bp element shows substantial structural similarities with other poxvirus promoters, providing further evidence that transcriptional elements are conserved within the poxviridae. the relative strengths of the lsdv early and late promoters were compared to the vaccinia virus (vv) p7.5k and p11k promoters in transient expression assays. these transient assays ...199910446656
a capripoxvirus detection pcr and antibody elisa based on the major antigen p32, the homolog of the vaccinia virus h3l gene.sheeppoxvirus (spv), goatpoxvirus (gpv) and lumpy skin disease virus (lsdv) of cattle belong to the capripoxvirus genus of the poxviridae family and can cause significant economic losses in countries where they are endemic. capripox diagnosis by classical virological methods dependent on live capripox virus is not suitable in countries such as australia where the virus is exotic and live virus is not available. to develop diagnostic tests based on recombinant material, we cloned and sequenced a ...199910485266
cdc42 and rac stimulate exocytosis of secretory granules by activating the ip(3)/calcium pathway in rbl-2h3 mast cells.we have expressed dominant-active and dominant-negative forms of the rho gtpases, cdc42 and rac, using vaccinia virus to evaluate the effects of these mutants on the signaling pathway leading to the degranulation of secretory granules in rbl-2h3 cells. dominant-active cdc42 and rac enhance antigen-stimulated secretion by about twofold, whereas the dominant-negative mutants significantly inhibit secretion. interestingly, treatment with the calcium ionophore, a23187, and the pkc activator, pma, re ...200010662774
detection and diagnosis of parapoxvirus by the polymerase chain reaction.the genus parapoxvirus includes four members, bovine papular stomatitis virus (bpsv), pseudocowpox virus (pcpv), orf virus (orfv) and parapoxvirus of red deer in new zealand (pvnz). a set of primers for polymerase chain reaction (pcr) was designed to detect viral dna from cells infected with each of the four parapoxviruses. the set of primers resulted in the amplification of appropriately sized products from cells infected with bpsv, pcpv, orfv and pvnz, respectively. the pcr method was applied ...200010680970
simple preparation of parapoxvirus genome dna for endonuclease analysis.we compared five methods for improved extraction of very-large parapoxvirus dna from infected cells: (i) alkaline-lysis procedure followed by phenol extraction; (ii) modified hirt procedure, which was a neutral lysis procedure followed by phenol extraction; (iii) hirt procedure; (iv) method used for extraction of vaccinia virus dna; and (v) standard procedure using virus purification with an ultracentrifuge and protease-sodium dodecyl sulfate-phenol treatment. the alkaline-lysis procedure was mo ...200010711601
human papillomavirus vaccines.genital human papillomavirus (hpv) infections are the viral sexually transmitted diseases most frequently diagnosed that include anogenital condylomas and squamous intra-$bepithelial lesions, among which the precursors of invasive carcinomas of the uterine cervix. in animal pv models, vaccination against l1 and/or l2 viral capsid proteins provides an efficient protection against infection, involving virus type-specific neutralizing antibodies. vaccination against non-structural e1, e2, e6 or e7 ...199910712890
bovine viral diarrhoea virus and bovine herpesvirus-1 prime uninfected macrophages for lipopolysaccharide-triggered apoptosis by interferon-dependent and -independent pathways.the flavivirus bovine viral diarrhoea (bvd) virus exists in two biotypes, cytopathic (cp) and non-cytopathic (ncp), defined by their effect on cultured cells. cp bvd virus-infected cells undergo apoptosis and may promote apoptosis in uninfected cells by an indirect mechanism. macrophages (m&phi;) infected with cp, but not ncp, bvd virus release a factor(s) in the supernatant capable of priming uninfected m&phi; for activation-induced apoptosis in response to lipopolysaccharide. a possible role o ...200010725412
human tumor growth is inhibited by a vaccinia virus carrying the e2 gene of bovine papillomavirus.papillomavirus is the etiologic agent associated with cervical carcinoma. the papilloma e2 protein is able to regulate negatively the expression of e6 and e7 papilloma oncoproteins. therefore, a new, highly attenuated vaccinia virus known as modified vaccinia virus ankara (mva), which carries the papillomavirus e2 gene, was used for the treatment of tumors associated with human papillomavirus.200010738224
poxvirus virions: their surface ultrastructure and interaction with the surface membrane of host cells.virions of vaccinia and orf viruses were examined by ultrahigh-resolution scanning electron microscopy using a non-coating method. intracellular mature particles of vaccinia virus appeared to be covered with a net and ultrastructurally their surface consists of many fine ridges and globules, while the surfaces of orf virus mature particles recovered from infected cells consist of spirally running protrusions. the ridge-like structures of vaccinia virus were presumed to correspond to surface tubu ...199910742959
long-term protective immunity to rinderpest in cattle following a single vaccination with a recombinant vaccinia virus expressing the virus haemagglutinin protein.a recombinant vaccine, produced by using a highly attenuated smallpox vaccine (lc16mo) as a vector and which expresses the rinderpest virus (rpv) haemagglutinin protein, has been developed. the properties of this vaccine, including its heat stability, efficacy in short-term trials, safety and genetic stability, have been confirmed in an earlier report. in the present study, the duration of the protective immunity generated by the vaccine in cattle was examined for up to 3 years following the adm ...200010811927
the fusion glycoprotein of human respiratory syncytial virus facilitates virus attachment and infectivity via an interaction with cellular heparan sulfate.human respiratory syncytial virus (rsv) f glycoprotein (rsv-f) can independently interact with immobilized heparin and facilitate both attachment to and infection of cells via an interaction with cellular heparan sulfate. rsv-glycosaminoglycan (gag) interactions were evaluated using heparin-agarose affinity chromatography. rsv-f from a2- and b1/cp-52 (cp-52)-infected cell lysates, rsv-f derived from a recombinant vaccinia virus, and affinity-purified f protein all bound to and were specifically ...200010864656
bpv1 e2 protein enhances packaging of full-length plasmid dna in bpv1 pseudovirions.we studied determinants of efficient encapsidation of circular dna, incorporating a pv early region dna sequence (nt 584-1978) previously shown to enhance packaging of dna within papillomavirus (pv)-like particles (vlps). insect coelomic cells (sf-9) and cultured monkey kidney cells (cos-1) were transfected with an 8-kb reporter plasmid incorporating the putative bpv packaging sequence and infected with bpv1 l1 and l2 recombinant baculovirus or vaccinia virus. heavy (1.34 g/ml) and light (1.30 g ...200010873782
independent modulation of collagen fibrillogenesis by decorin and lumican.the leucine-rich proteoglycans (also known as "small, leucine-rich proteoglycans," or slrps) lumican and decorin are thought to be involved in the regulation of collagen fibril assembly. preparation of these proteoglycans in chemical amounts without exposure to denaturants has recently been achieved by infecting ht-1080 cells with vaccinia virus that contains an expression cassette for these molecules. addition of lumican and decorin to a collagen fibrillogenesis assay based on turbidity demonst ...200010892350
new tuberculosis vaccines based on attenuated strains of the mycobacterium tuberculosis complex.the world urgently needs a better tuberculosis vaccine. bacille calmette-guerin (bcg), an attenuated strain of mycobacterium bovis, has been very widely used as a vaccine for many years but has had no major effect on reducing the incidence of tuberculosis. a number of alternative living and non-living vaccines are being investigated. live vaccine candidates include genetically modified forms of bcg, genetically attenuated strains of the mycobacterium tuberculosis complex and genetically engineer ...200010947858
gammadelta(+) t-lp6phocyte cytotoxicity against envelope-expressing target cells is unique to the alymphocytic state of bovine leukemia virus infection in the natural host.bovine leukemia virus (blv) is a complex b-lymphotrophic retrovirus of cattle and the causative agent of enzootic bovine leukosis. serum antibody in infected animals does not correlate with protection from disease, yet only some animals develop severe disease. while a cytotoxic t-lymphocyte response may be responsible for directing blv pathogenesis, this possibility has been left largely unexplored, in part since the lack of readily established cytotoxic target cells in cattle has hampered such ...200010954528
improved technique for transient expression and negative strand virus rescue using fowlpox t7 recombinant virus in mammalian cells.the suitability of recombinant t7 polymerase produced using either the highly attenuated mva strain of vaccinia (mva-t7) or fowlpox virus (fp-t7) for transient expression and negative strand virus rescue was compared in two mammalian cell lines (mdbk and vero) and in primary cells of bovine, ovine and caprine origin. such primary cells are more permissive for the growth of wild type strains of morbilliviruses, such as rinderpest virus and peste des petits ruminants virus. mva-t7 was found to be ...200010996645
a recombinant vaccinia virus containing the papilloma e2 protein promotes tumor regression by stimulating macrophage antibody-dependent cytotoxicity.human papillomavirus infection is associated with cervical cancer. the e6 and e7 papillomavirus proteins are normally required for the maintenance of the malignant phenotype. expression of these proteins in infected cells is negatively regulated by the binding of the papilloma e2 protein to the long terminal control region of the papilloma virus genome. the e2 protein can also promote cell arrest and apoptosis in hela cells. therefore, it is clear that this protein has the potential of inhibitin ...200010999461
an emergent poxvirus from humans and cattle in rio de janeiro state: cantagalo virus may derive from brazilian smallpox vaccine.the biological properties of poxvirus isolates from skin lesions on dairy cows and milkers during recent exanthem episodes in cantagalo county, rio de janeiro state, brazil, were more like vaccinia virus (vv) than cowpox virus. pcr amplification of the hemagglutinin (ha) gene substantiated the isolate classification as an old world orthopoxvirus, and alignment of the ha sequences with those of other orthopoxviruses indicated that all the isolates represented a single strain of vv, which we have ...200011080491
delivery of neospora caninum surface protein, ncsrs2 (nc-p43), to mouse using recombinant vaccinia order to develop a vaccine against neospora caninum in dogs and cattle, we constructed a recombinant vaccinia virus expressing the n. caninum surface protein, ncsrs2 (nc-p43). monoclonal antibodies to ncsrs2 and anti-n. caninum tachyzoite mouse serum recognized the ncsrs2 expressed by the recombinant vaccinia virus. in addition, recombinant ncsrs2 was transported to the cell surface. mice infected with the recombinant virus predominantly produced igg1 antibody (ab) to n. caninum, rather than ...200011097303
spectrum of virus inhibition by consensus interferon ym643.the spectrum of viruses inhibited by a genetically engineered consensus interferon (ifn) ym643 (interferon alfacon-1) was evaluated using a cytopathic effect inhibition assay or plaque inhibition assay for five dna viruses and 12 rna viruses. this activity was compared to that of natural ifn-alpha derived from namalwa lymphoblastoid cell line [ifn-alpha (namalwa)]. the viruses inhibited by both ifns were herpesvirus types 1 and 2, human cytomegalovirus, varicella-zoster virus, vesicular stomatit ...200011142632
prevention of vertical transmission of neospora caninum in balb/c mice by recombinant vaccinia virus carrying ncsrs2 gene.neospora caninum infection is the major cause of bovine abortion. to develop a vaccine against n. caninum infection, recombinant vaccinia viruses carrying ncsrs2 and ncsag1 genes (vv/nc-p43 and vv/nc-p36, respectively) were constructed and were tested in a mouse model. vaccination of dams with vv/nc-p43 appeared to confer effective protection against vertical transmission to offspring, though that with vv/nc-p36 only provided partial protection. moreover, the vv/nc-p43 vaccination provoked cellu ...200111166895
glycosaminoglycan binding properties of the myxoma virus cc-chemokine inhibitor, m-t1.poxviruses encode a number of secreted virulence factors that function to mitigate or modulate the host immune response. m-t1 is a secreted 43-kda glycoprotein produced by the myxoma virus, a poxvirus pathogen of rabbits, that binds cc-chemokines with high affinity, blocks binding to their cognate g-protein coupled receptors, and thereby inhibits chemokine-induced leukocyte chemotaxis. the present study indicates that m-t1, but not the related vaccinia virus 35-kda cc-chemokine-binding protein, ...200111369757
[angiogenin and its role in angiogenesis].the review is devoted to angiogenin, one of the factors that induce formation of blood vessels, which is unique among them in that it is a ribonuclease. consideration is given to the tertiary structure of human angiogenin; the catalytic and cell-receptor binding sites, their significance for angiogenic activity; the human angiogenin gene structure, chromosomal localization, and expression; the specificity of angiogenin as a ribonuclease and abolishment of protein synthesis; the nuclear localizat ...200111443914
efficiency of delivery of dna to cells by bovine papillomavirus type-1 l1/l2 investigate the efficiency of encapsidation of plasmid by papillomavirus virus-like particles (pv vlps), and the infectivity of the resultant pv pseudovirions, cos-1 cells were transfected with an 8-kb plasmid incorporating a green fluorescent protein (gfp) reporter gene (pgsv), and infected with bovine pv (bpv-1) l1/l2 recombinant vaccinia virus to produce bpv1 pseudovirions. approximately 1 in 1.5 x 10(4) of dense (1.35 g/ml) pv pseudovirions and 0.3 in 10(4) of less-dense (1.29 g/ml) pseud ...200111499923
rescue of bovine respiratory syncytial virus from cloned cdna: entire genome sequence of brsv strain a51908.infectious bovine respiratory syncytial virus (brsv) was produced by intracellular co-expression of five plasmid borne cdnas, each under the control of a t7 rna polymerase promoter. these separately encoded a full-length, genetically-marked copy of brsv antigenome along with either brsv or human respiratory syncytial virus (hrsv) support plasmids, which express n, p, l and m2-1 proteins. hep2 cells were used in transfection and recombinant vaccinia virus (mva-t7) provided t7 rna polymerase to dr ...200111724268
long-term sterilizing immunity to rinderpest in cattle vaccinated with a recombinant vaccinia virus expressing high levels of the fusion and hemagglutinin glycoproteins.rinderpest is an acute and highly contagious viral disease of ruminants, often resulting in greater than 90% mortality. we have constructed a recombinant vaccinia virus vaccine (v2rvfh) that expresses both the fusion (f) and hemagglutinin (h) genes of rinderpest virus (rpv) under strong synthetic vaccinia virus promoters. v2rvfh-infected cells express high levels of the f and h glycoproteins and show extensive syncytium formation. cattle vaccinated intramuscularly with as little as 10(3) pfu of ...200211752138
[smallpox and smallpox virus--200 years since the first vaccination in norway].in december 1801, the first vaccination against smallpox in norway took place. vaccine material came from denmark, england, ireland, and other countries; it was also obtained from a few local cowpox cases. what mattered was the effect, not the origin. several reports indicate that variola virus itself, the cause of smallpox, was also used for human vaccination after passages through cows and horses. a vaccine institute for production of vaccine in calves was established in kristiania in 1891. co ...200111808015
characterization of wild-type and cidofovir-resistant strains of camelpox, cowpox, monkeypox, and vaccinia viruses.cidofovir ([(s)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine] [hpmpc])-resistant forms of camelpox, cowpox, monkeypox, and vaccinia viruses were developed by prolonged passage in vero 76 cells in the presence of drug. eight- to 27-fold-higher concentrations of cidofovir were required to inhibit the resistant viruses than were needed to inhibit the wild-type (wt) viruses. resistant viruses were characterized by determining their cross-resistance to other antiviral compounds, examining their di ...200211959564
the genomes of sheeppox and goatpox viruses.sheeppox virus (sppv) and goatpox virus (gtpv), members of the capripoxvirus genus of the poxviridae, are etiologic agents of important diseases of sheep and goats in northern and central africa, southwest and central asia, and the indian subcontinent. here we report the genomic sequence and comparative analysis of five sppv and gtpv isolates, including three pathogenic field isolates and two attenuated vaccine viruses. sppv and gtpv genomes are approximately 150 kbp and are strikingly similar t ...200212021338
detection of smallpox virus dna by lightcycler pcr.a 300-bp plasmid fragment of the hemagglutinin gene was used as target dna to develop a rapid real-time lightcycler (roche applied science, indianapolis, ind.) pcr assay for laboratory detection of smallpox virus. pcr primers and probes were designed specifically for detection of smallpox virus dna, but all viruses of the genus orthopoxvirus tested could be detected by use of the hemagglutinin gene target sequence. base pair mismatches in the 204-bp amplicon allowed discrimination of cowpox viru ...200212037052
continuous porcine cell lines developed from alveolar macrophages: partial characterization and virus susceptibility.porcine monomyeloid cell lines were established following transfection of primary porcine alveolar macrophage cultures with plasmid psv3neo, carrying genes for neomycin resistance and sv40 large t antigen. the parental clone 3d4 exhibited a relatively rapid doubling time (25.5 h), high plating efficiency and mixed phenotype with respect to growth on a solid support. single cell cloning of the 3d4 parent resulted in establishment of several cell lines; three of them designated 3d4/2, 3d4/21 and 3 ...200212088830
the virucidal spectrum of a high concentration alcohol mixture.the virucidal spectrum of a high concentration alcohol mixture (80% ethanol and 5% isopropanol) was determined for a broad series of lipid-enveloped (le) and non-lipid-enveloped (nle) viruses covering all relevant blood-borne viruses. le viruses were represented by human immunodeficiency virus (hiv), bovine viral diarrhoea virus (bvdv), a specific model virus for hepatitis c virus (hcv), pseudorabies virus (prv), and vaccinia virus. for the nle viruses hepatitis a virus, canine parvovirus (a mod ...200212090799
recognition of bovine respiratory syncytial virus proteins by bovine cd8+ t lymphocytes.cd8+ t lymphocytes play a major role in the clearance of bovine respiratory syncytial virus (brsv), an important respiratory pathogen of young calves that shares many of the epidemiological and pathological features of human respiratory syncytial virus (hrsv) in infants. recombinant vaccinia virus (rvv) and recombinant fowlpox virus (rfpv), expressing individual brsv proteins, were used to demonstrate that the f, n and m2 proteins were the major antigens recognized by bovine cd8+ t cells in majo ...200312562331
araçatuba virus: a vaccinialike virus associated with infection in humans and cattle.we describe a vaccinialike virus, araçatuba virus, associated with a cowpoxlike outbreak in a dairy herd and a related case of human infection. diagnosis was based on virus growth characteristics, electron microscopy, and molecular biology techniques. molecular characterization of the virus was done by using polymerase chain reaction amplification, cloning, and dna sequencing of conserved orthopoxvirus genes such as the vaccinia growth factor (vgf), thymidine kinase (tk), and hemagglutinin. we u ...200312603984
cell-mediated cytotoxicity in recovery from poxvirus infections.the availability of mutant and gene targeted knockout mice with defects in components of cellular cytotoxicity mediated by either the fas or the exocytosis pathway permitted an analysis of their role in recovery from poxvirus infections. ectromelia (ev), a natural mouse pathogen causing mousepox, the closely related orthopoxviruses cow pox (cpv) and vaccinia virus (vv), each encode serpins that inhibit fas mediated apoptosis and lysis of target cells. nevertheless, distinct differences were seen ...200312820184
[the demonstration of vaccinia virus in the spinal cord of young cattle]. 195213006751
milker's nodules: isolation of a poxvirus from a human case.a virus of the pox group was isolated from a lesion on a patient with milker's nodules by use of tissue cultures of bovine cells. the virus shows marked biologic and serologic differences from vaccinia virus, and it is presumably the etiologic agent of the milker's nodule syndrome.196313959825
isolation of a viral agent from pseudocowpox disease.a virus with characteristics similar to the pox group was isolated in tissue culture from bovine skin and oral lesions typical of pseudocowpox. the properties of this agent, certain of which differ from vaccinia virus, suggest that the isolate is the etiologic agent of the pseudocowpox syndrome.196314043336
expression vehicles used in recombinant dna technology.we survey cloning vehicles whose function is to carry and express a gene in host cells including escherichia coli, saccharomyces cerevisiae and mammalian cells. in e. coli these include vehicles based on the lac operon, the trp operon, the rho leftward operon, and the reca gone; open reading frame cloning vehicles are also discussed, as are steps that can be taken to extrude a gene product from the cell and the use of plasmids with runaway replication. in s. cerevisiae we discuss vehicles based ...198314540892
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