TitleAbstractYear(sorted ascending)
an equine herpesvirus type 1 recombinant with a deletion in the ge and gi genes is avirulent in young horses.the cell culture-adapted kya strain of equine herpesvirus type 1 (ehv-1) has been found to be attenuated in young horses (matsumura et al., 1996, vet. microbiol. 48, 353-365). the kya strain lacks at least six genes in its genome, including those encoding glycoproteins ge and gi. to elucidate whether ehv-1 glycoproteins ge and gi play a role in viral virulence, we have constructed an ehv-1 recombinant that has the genes encoding both ge and gi deleted from its genome and its revertant. growth pr ...19989501037
protective effects of equine herpesvirus-1 (ehv-1) glycoprotein b in a murine model of ehv-1-induced an attempt to determine if pregnant mice could be protected from abortion subsequent to challenge with equine herpesvirus-1 (ehv-1) in the mouse model of ehv-1 disease, female balb/c mice were inoculated with baculovirus-expressed ehv-1 glycoprotein b (bac-gb), wild-type baculovirus (bac-wt), rabbit kidney (rk-13) or baby hamster kidney (bhk-21) cells. using an elisa, antibodies against ehv-1 were detected in the serum of mice following two injections of bac-gb and were enhanced by a third in ...19989791876
the equine herpesvirus 1 ir6 protein that colocalizes with nuclear lamins is involved in nucleocapsid egress and migrates from cell to cell independently of virus infection.the equine herpesvirus 1 (ehv-1) ir6 protein forms typical rod-like structures in infected cells, influences virus growth at elevated temperatures, and determines the virulence of ehv-1 rac strains (osterrieder et al., virology 226:243-251, 1996). experiments to further elucidate the functions and properties of the ir6 protein were conducted. it was shown that the ir6 protein of wild-type racl11 virus colocalizes with nuclear lamins very late in infection as demonstrated by confocal laser scan m ...19989811716
diagnosis and sero-epizootiology of equine herpesvirus type 1 and type 4 infections in japan using a type-specific elisa.recently, a type-specific elisa using equine herpesvirus type 1 (ehv-1) and type 4 (ehv-4) glycoprotein gs (ggs) was developed by crabb and studdert [1993]. to investigate the dissemination of ehv-1 and -4 among horses in japan, we applied their elisa as suitable for discriminating between ehv-1 and -4 infections serologically. type-specificity of the elisa was confirmed by using paired sera of infected horses with either ehv-1 or -4. application of the elisa to sera collected before and after t ...19989819768
epidemiology of equine herpesvirus abortion: searching for clues to the future. 19989830567
seroprevalence of equine herpesvirus 1 in thoroughbred foals before and after investigate the seroprevalence of equine herpesvirus 1 in foals around weaning and after weaning on two large thoroughbred farms using a type-specific enzyme-linked immunosorbent assay to determine exposure to infection.19989830568
systemic infection by equid herpesvirus-1 in a grevy's zebra stallion (equus grevyi) with particular reference to genital pathology.a severe multi-systemic form of equid herpesvirus-1 infection is described in an adult zebra stallion. there was multifocal necrotizing rhinitis, marked hydrothorax and pulmonary oedema, with viral antigen expression in degenerating epithelial cells, local endothelial cells and intravascular leucocytes of the nasal mucosa and lung. specific localization of ehv-1 infection was seen in the testes and epididymides, including infection of leydig cells and germinal epithelium, which would have facili ...19989839210
deletion of multiple immediate-early genes from herpes simplex virus reduces cytotoxicity and permits long-term gene expression in neurons.herpes simplex virus type 1 (hsv-1) has many attractive features that suggest its utility for gene transfer to neurons. however, viral cytotoxicity and transient transgene expression limit practical applications even in the absence of viral replication. mutant viruses deleted for the immediate early (ie) gene, icp4, an essential transcriptional transactivator, are toxic to many cell types in culture in which only the remaining ie genes are expressed. in order to test directly the toxicity of oth ...199810023438
primary and challenge infection of mice with equine herpesvirus 1, strain hsv25a.clinical signs, haematology, lymphocyte subset analysis, viral clearance, lung histopathology and humoral and cell-mediated (cmi) immune responses were monitored throughout the acute and convalescent phases of infection in groups of balb/c mice infected intranasally with equine herpesvirus 1 (ehv-1), strain hsv25a. primary infection caused a leucocytosis due to a neutrophilia during days 1 and 2 post-infection (pi) and a b lymphocytosis at day 1 pi. serum elisa antibodies were detected by 7 days ...19989870583
an equine herpesvirus 1 mutant with a lacz insertion between open reading frames 62 and 63 is replication competent and causes disease in the murine respiratory equine herpesvirus 1 (ehv-1) mutant was constructed by inserting a lacz expression cassette into the intergenic region upstream of gene 62 (glycoprotein l; gl) and downstream of gene 63 (a homologue of the herpes simplex virus transcriptional activator icp0). the recombinant lacz62/63-ehv-1 had similar growth kinetics in cell culture to those of the parental wild type (wt) virus, with indistinguishable cytopathic effects and plaque morphology. reverse transcriptase pcr confirmed that the lacz ...19989856103
equine herpesvirus-4 glycoprotein g is secreted as a disulphide-linked homodimer and is present as two homodimeric species in the virion.glycoprotein g (gg) homologues have been found in most alphaherpesviruses although little is known about their structure or function. in this study, three species of equine herpesvirus-4 (ehv-4) gg were identified: a full-length 68 kda virion-associated species (ggvl), a 12 kda virion-associated species (ggvs) and a 60 kda secreted species (ggs), detected in the medium of infected cells. ggs and ggvs appear to be proteolytic cleavage products of ggvl and correspond to the n- and c-terminal regio ...19989603336
genomic organization of the canine herpesvirus us region.canine herpesvirus (chv) is an alpha-herpesvirus of limited pathogenicity in healthy adult dogs and infectivity of the virus appears to be largely limited to cells of canine origin. chv's low virulence and species specificity make it an attractive candidate for a recombinant vaccine vector to protect dogs against a variety of pathogens. as part of the analysis of the chv genome, the authors determined the complete nucleotide sequence of the chv us region as well as portions of the flanking inver ...19989620207
nucleotide sequences of glycoprotein i and e genes of equine herpesvirus type 4.the nucleotide sequences of the glycoprotein i (gi) and e (ge) genes of equine herpesvirus type 4 (ehv-4) strain th20 were determined. the predicted region encoding the ehv-4 gi gene is 1,263 nucleotides, corresponding to a polypeptide of 420 amino acids in length. the predicted region encoding the ehv-4 ge gene is 1,647 nucleotides, corresponding to a polypeptide of 548 amino acids in length. the ehv-4 gi and ge genes show 74% and 85% identity at the amino acid level with those of equine herpes ...19989524947
the bovine herpesvirus type 1 ul3.5 open reading frame encodes a virion structural protein.the bovine herpesvirus type 1 (bhv-1) open reading frame (orf) ul3.5 is similar to orfs found in pseudorabies virus, infectious laryngotracheitis virus, equine herpesvirus type 1, and varicella zoster virus, but clearly absent from herpes simplex virus. the published sequence for this orf predicts a 126-amino-acid (13.2 kda) protein product with an isoelectric point of 12.3. we confirmed the ul3.5 sequence, expressed the orf as a glutathione-s-transferase fusion protein, and made rabbit antibodi ...19989448691
health evaluation of free-ranging guanaco (lama guanicoe).twenty free-ranging guanaco (lama guanicoe) in chubut province, argentina, were immobilized for health evaluations. all but two animals appeared to be in good condition. hematology, serum chemistry, and vitamin and mineral levels were measured, and feces were evaluated for parasites. serology tests included bluetongue, brucellosis, bovine respiratory syncitial virus, bovine viral diarrhea/mucosal disease, equine herpesvirus 1, infectious bovine rhinotracheitis, johne's disease (mycobacterium par ...19989732026
the equid herpesvirus-1 gene 63 is expressed as a leaky late (gamma 1) transcript and is nonessential for replication in vitro.the regulation of equid herpesvirus-1 (ehv-1) gene 63 was investigated using molecular expression studies and its role in viral growth was identified by constructing a gene 63 mutant virus. metabolic inhibitors were used to show that ehv-1 gene 63 is expressed as a leaky-late (gamma 1) transcript. transient transfections and subsequent chloramphenicol acetyltransferase (cat) reporter assays showed that gene 63 was transactivated by ehv-1 gene 64 (immediate early) protein. an ehv-1 gene 63 mutant ...19989696126
phosphorylation of structural components promotes dissociation of the herpes simplex virus type 1 tegument.the role of phosphorylation in the dissociation of structural components of the herpes simplex virus type 1 (hsv-1) tegument was investigated, using an in vitro assay. addition of physiological concentrations of atp and magnesium to wild-type virions in the presence of detergent promoted the release of vp13/14 and vp22. vp1/2 and the ul13 protein kinase were not significantly solubilized. however, using a virus with an inactivated ul13 protein, we found that the release of vp22 was severely impa ...19989696804
diversity of genomic electropherotypes of naturally occurring equine herpesvirus 1 isolates in argentina.the genomes of 10 equine herpesvirus 1 (ehv-1) strains isolated in argentina from 1979 to 1991, and a japanese hh1 reference strain were compared by restriction endonuclease analysis. two restriction enzymes, bamhi and bglii, were used and analysis of the electropherotypes did not show significant differences among isolates obtained from horses with different clinical signs. this suggests that the ehv-1 isolates studied, which circulated in argentina for more than 10 years, belong to a single ge ...19989698821
monitoring and detection of acute viral respiratory tract disease in develop a system to monitor and detect acute infections of the upper respiratory tract (i.e., nares, nasopharynx, and pharynx) in horses and to assess the association among specific viral infections, risk factors, and clinical signs of disease.19989702229
cosolvents facilitate dna synthesis in the herpes simplex virus 1 unique short (us) inverted repeat.dna synthesis under standard conditions is not successful within a portion of the us1 gene of hsv-1 which is juxtaposed to an 86% g + c-containing tract in the us inverted repeat sequence. we report that the independent addition of specific amounts of at least three different types of cosolvents is capable of facilitating dna synthesis within this g + c-rich region. in addition, this strategy was used to successfully place a specific site-directed mutation in the us1 gene. consideration of these ...19989705174
complementation of a replication-defective mutant of equine herpesvirus type 1 by a cell line expressing the immediate-early protein.equine herpesvirus type 1 (ehv-1) possesses a sole, diploid immediate-early (ie) gene that encodes a major regulatory protein of 1487 amino acids capable of modulating expression of both early and late ehv-1 promoters and capable of trans-repressing its own promoter. in this study, a rabbit kidney cell line (ie13.1) that constitutively expresses the ehv-1 ie protein was generated by cotransfection of rabbit kidney (rk-13) cells with the viral ie gene and a neomycin resistance marker. the ie prot ...19989705258
herpesvirus myeloencephalopathy in horses: 11 cases (1982-1996).to determine results of csf analysis in horses with equid herpesvirus myeloencephalopathy (ehm) and to determine whether results of csf analysis were associated with outcome.19989731262
cellular transcription factors enhance herpes simplex virus type 1 oris-dependent dna replication.the herpes simplex virus type 1 (hsv-1) origin of dna replication, oris, contains three binding sites for the viral origin binding protein (obp) flanked by transcriptional regulatory elements of the immediate-early genes encoding icp4 and icp22/47. to assess the role of flanking sequences in oris function, plasmids containing oris and either wild-type or mutant flanking sequences were tested in transient dna replication assays. although the icp4 and icp22/47 regulatory regions were shown to enha ...19989557644
study of the protective immunity of co-expressed glycoprotein h and l of equine herpesvirus-1 in a murine intranasal infection model.equine herpesvirus-1 (ehv-1) glycoproteins h, and l (gh and gl) expressed individually or co-expressed by recombinant baculoviruses were used to immunise balb/c mice prior to intranasal challenge in a murine model of respiratory infection. only the co-expressed material (ehv-1 gh/gl) induced neutralising antibody (low levels). the same immunogen also produced the strongest cellular responses. immunisation with gh/gl and, to a lesser extent, with gh alone was associated with a reduction of virus ...19989595623
the dna sequence of equine herpesvirus-4.the complete dna sequence of equine herpesvirus-4 (ehv-4) strain ns80567 was determined. the genome is 145597 bp in size and consists of a long unique region (ul, 112398 bp) flanked by a short inverted repeat (trl/irl, 27 bp) linked to a short unique region (us, 12789 bp) flanked by a substantial inverted repeat (trs/irs, 10178 bp). ehv-4 is predicted to contain 76 different genes; three of these are present twice in trs/irs, giving a total of 79 genes. the closely related virus equine herpesvir ...19989603335
characterization of the cytolytic t-lymphocyte response to a candidate vaccine strain of equine herpesvirus 1 in cba mice.the cytolytic t-lymphocyte (ctl) response to respiratory infection with equine herpesvirus 1 (ehv-1) in cba (h-2(k)) mice was investigated. intranasal (i.n.) inoculation of mice with the attenuated ehv-1 strain kya resulted in the generation of a primary virus-specific ctl response in the draining mediastinal lymph nodes 5 days following infection. ehv-1-specific ctl could be restimulated from the spleen up to 26 weeks after the resolution of infection, indicating that a long-lived memory ctl po ...19989620990
in situ study on the pathogenesis and immune reaction of equine herpesvirus type 1 (ehv-1) infections in mice.the mouse model was used to study the pathogenesis of equine herpesvirus type 1 (ehv-1) after primary and secondary intranasal infections. within a few hours after infection, ehv-1 was found in nasal and olfactorial epithelium and sub-epithelial cells of the respiratory mucosa, but antigen-specific immune cells were never detected. next to the lung, ehv-1 was transmitted early and directly to the brain, both via the olfactory route and the trigeminal nerve, but traces of degenerative or inflamma ...19989640242
in vitro reactivation of latent equid herpesvirus-1 from cd5+/cd8+ leukocytes indirectly by il-2 or chorionic and equine chorionic gonadotrophin (ecg) initiated reactivation of equid herpesvirus-1 (ehv-1) from venous lymphocytes at a frequency of 1/10(-5). indirect immunofluorescence showed that > 80% of virus-positive leukocytes were cd5+/cd8+ with the remaining 20% being cd5+/cd8-/cd4-. cocultivation demonstrated that the reactivated virus was infectious. in addition, virus was reactivated in vitro from leukocytes of > 70% of horses by the mitogens phytohaemagglutinin (pha) and pokeweed mitogen ( ...19989880014
dendritic cells presenting equine herpesvirus-1 antigens induce protective anti-viral immunity.equine herpesvirus-1 (ehv-1) causes rhinopneumonitis, abortion and cns disorders in horses. using intranasal inoculation, the mouse model of this disease mimics the major pathogenic and clinical features of the equine disease. the aim of this study was to investigate whether murine dendritic cells (dc) can be infected with ehv-1 and whether they can be used as cellular vaccines for the induction of prophylactic anti-ehv-1 immunity. it was found that the dc lines fsdc, d2sc1, 18 (all h-2d) and 80 ...19989880015
immune responses and protective efficacy of recombinant baculovirus-expressed glycoproteins of equine herpesvirus 1 (ehv-1) gb, gc and gd alone or in combinations in balb/c mice.baculovirus-expressed glycoproteins of ehv-1 gb, gc and gd alone or in combination evoked antibody responses and protected vaccinated mice against a challenge with ehv-1. gb, gd, gb + gc, gb + gd and gc + gd elicited very high levels of elisa antibodies while gc and gc + gd elicited high levels of virus neutralising antibodies. western blotting demonstrated that the antibodies produced were not only specific for the baculovirus-expressed glycoproteins gb, gc and gd, but also highly specific for ...19989646476
preparation of hsv-dna and production of infectious virus.this chapter deals with (1) the preparation of herpes simplex virus (hsv) virion dna of a quality and purity suitable to be used for the generation of infectious virus, and (2) its use in the preparation of infectious virus. an important development in the understanding of virus genetics and gene products has been the ability to carry out reverse genetics. this is dependent on the ability to manipulate the genome in vitro and reconstitute infectious virus. our understanding of dna viruses and po ...199821374220
the equine herpesvirus 1 us2 homolog encodes a nonessential membrane-associated virion component.experiments were conducted to analyze the equine herpesvirus 1 (ehv-1) gene 68 product which is encoded by the ehv-1 us2 homolog. an antiserum directed against the amino-terminal 206 amino acids of the ehv-1 us2 protein specifically detected a protein with an mr of 34,000 in cells infected with ehv-1 strain racl11. ehv-1 strain ab4 encodes a 44,000-mr us2 protein, whereas vaccine strain rach, a high-passage derivative of racl11, encodes a 31,000-mr us2 polypeptide. irrespective of its size, the ...199910074198
equid herpesvirus-induced immunosuppression is associated with lymphoid cells and not soluble circulating factors.a paresis isolate of equid herpesvirus 1 (ehv1, ab4/8) and a plaque-purified virus derived from it (ehv1, ab4/13), induced long-term suppression of both mitogenic and antigen-specific lymphocyte proliferations in adult outbred ponies. peripheral blood mononuclear cells (pbmc) taken from a pony after ehv1 infection suppressed the in vitro function of normal cells but serum did not. this showed that the observed immune suppression was associated with circulating pbmc and/or their products rather t ...199910630790
equine herpesvirus type 1 infects dendritic cells in vitro: stimulation of t lymphocyte proliferation and cytotoxicity by infected dendritic cells.equine herpesvirus type 1 (ehv-1) causes respiratory disease, abortion and myeloencephalopathy in horses. as with other herpesviruses, cell-mediated immunity is considered important for both recovery and protection. although virus-specific t-cell proliferation and cytotoxicity can be detected following in vivo infection, little is known about the role of antigen presenting cells such as dendritic cells (dcs) in these processes. peripheral blood dcs were shown to express the viral glycoprotein gb ...19999950351
clinical, haematological and biochemical findings in foals with neonatal equine herpesvirus-1 infection compared with septic and premature foals.a retrospective multicentre study comparing historical, clinical, haematological, acid-base and biochemical findings of foals with equine herpesvirus-1 (ehv-1) infection, septicaemia or prematurity was performed to determine if early diagnosis of ehv-1 foals was possible. fifty-three foals were studied and were assigned to one of 2 groups: herpes positive (n = 14) or herpes negative (n = 39). the latter group included 20 septic, 11 premature, and 8 premature and septic foals. the presence of her ...199910505959
induction and prevention of apoptosis in human hep-2 cells by herpes simplex virus type 1.cultured human epithelial cells infected with an icp27 deletion strain of herpes simplex virus type 1 (hsv-1) show characteristic features of apoptotic cells including cell shrinkage, nuclear condensation, and dna fragmentation. these cells do not show such apoptotic features when infected with a wild-type virus unless the infections are performed in the presence of a protein synthesis inhibitor. thus, both types of virus induce apoptosis, but the icp27-null virus is unable to prevent this proce ...199910559354
[mutations in the us2 and glycoprotein b genes of the equine herpesvirus 1 vaccine strain rach have no effects on its attenuation].the equine herpesvirus 1 (ehv-1) modified live vaccine strain rach is apathogenic for both laboratory animals and the natural host. the apathogenicity of rach was caused by serial passages of the virus in heterologous cells. when compared to the virulent parental strain racl11 several changes in the rach genome occurred. previous results have shown that the loss of the ir6 gene correlated with the loss of virulence. additional important mutations were observed within the us2 gene which is direct ...199910507185
determination of equid herpesvirus 1-specific, cd8+, cytotoxic t lymphocyte precursor frequencies in ponies.the frequency of antigen-specific, genetically restricted cytotoxic t lymphocyte precursors (ctlp) was measured in peripheral blood mononuclear cells (pbmc) of ponies before and after infection with equid herpesvirus 1 (ehv1). split-well limiting dilution analysis (lda) was developed to measure ctlp frequency using ehv1-infected 51cr-labelled lymphoblasts as targets. extensive characterisation showed that recombinant human interleukin-2, autologous antigen presenting cells and equine serum conta ...199910507286
equine herpes virus type 1 (ehv-1) infection induces alterations in the cytoskeleton of vero cells but not apoptosis.effects of infection with two different strains of equine herpes virus type 1 (ehv-1; piber 178/83, kentucky d) on the cytoskeleton of vero cells were investigated immunohistochemically, and evaluated by confocal laser scanning microscopy. twenty four hours post ehv-1 infection the assembly of the microtubulus system of vero cells was heavily disturbed. the golgi region was dispersed into vesicles spread throughout the cytoplasm as demonstrated by wga lectin binding. other cytoskeletal elements ...199910542029
the gamma2 late glycoprotein k promoter of equine herpesvirus 1 is differentially regulated by the ie and eicp0 proteins.the equine herpesvirus 1 immediate-early (ie) phosphoprotein is essential for the activation of transcription from viral early and late promoters and trans-represses its own promoter. transient-transfection assays showed that the ie protein trans-represses the gamma2 late gk promoter. gel shift and dnase i footprinting assays demonstrated that the ie protein binds to the gk promoter sequences from -42 to -26 and from -13 to +12 that overlap the transcription initiation site (+1). these results i ...199910191181
immunogenicity of herpes simplex virus type 1 mutants containing deletions in one or more alpha-genes: icp4, icp27, icp22, and icp0.replication defective mutants of hsv have been proposed both as vaccine candidates and as vehicles for gene therapy because of their inability to produce infectious progeny. the immunogenicity of these hsv replication mutants, at both qualitative and quantitative levels, will directly determine their effectiveness for either of these applications. we have previously reported (brehm et al., j. virol., 71, 3534, 1997) that a replication defective mutant of hsv-1, which expresses a substantial leve ...199910191191
the nucleotide sequence of the glycoprotein g homologue of equine herpesvirus 3 (ehv3) indicates ehv3 is a distinct equid alphaherpesvirus.ehv3 causes equine coital exanthema and has been classified as an alphaherpesvirus on the basis of its biological properties; however due to the absence of any sequence information the phylogenetic relationship has not previously been examined. the complete nucleotide sequence of the ehv3 glycoprotein g (gg) gene was determined and showed that this virus is most closely related to the alphaherpesviruses equine herpesviruses type 1 (ehv 1) and type 4 (ehv4). ehv3 gg contains conserved and variabl ...199910550674
the defective interfering particles of equine herpesvirus 1 encode an icp22/icp27 hybrid protein that alters viral gene regulation.the genomes of equine herpesvirus 1 (ehv-1) defective interfering (di) particles that mediate persistent infection were shown to encode a unique hybrid open reading frame composed of sequences that encode the 196 n-terminal amino acids of icp22 linked in-frame to the c-terminal 68 amino acids of icp27. previous studies demonstrated that this hybrid gene, designated as icp22/icp27. was expressed abundantly at both the mrna and the protein levels in di particle-enriched infections, but not in stan ...199910082387
construction and characterization of an equine herpesvirus 1 glycoprotein c negative equine herpesvirus 1 (ehv-1) strain racl 11 mutant was constructed that carries the escherichia coli lacz gene instead of the open reading frame encoding glycoprotein c (gc). the engineered virus mutant (l11(delta)gc) lacked codons 46-440 of the 1404 bp gene. on rabbit kidney cell line rk13 and equine dermal cell line edmin337, the l11(delta)gc virus grew to titers which were reduced by approximately 5- to 10-fold compared with wild-type racl11 virus or a repaired virus (r-l11(delta)gc). howe ...199910082388
epidemiology of ehv-1 and ehv-4 in the mare and foal populations on a hunter valley stud farm: are mares the source of ehv-1 for unweaned foals.the prevalence of ehv-1 and ehv-4 antibody-positive horses was determined using a type specific elisa on serum samples collected from 229 mares and their foals resident on a large thoroughbred stud farm in the hunter valley of new south wales in february 1995. more than 99% of all mares and foals tested were ehv-4 antibody positive, while the prevalence of ehv-1 antibody positive mares and foals were 26.2 and 11.4%, respectively. examination of the elisa absorbance data for the individual mares ...199910501159
dna-mediated immunization with glycoprotein d of equine herpesvirus 1 (ehv-1) in a murine model of ehv-1 respiratory infection.dna-mediated immunization was assessed in a murine model of equine herpesvirus 1 (ehv-1) respiratory infection. a single intramuscular injection with plasmid dna encoding ehv-1 glycoprotein d (ehv-1 gd), including its predicted c-terminal membrane anchor sequence, induced a specific antibody response detectable by 2 weeks and maintained through 23 weeks post injection. a second injection at 4 weeks markedly enhanced the antibody response and all ehv-1 gd-injected mice developed neutralizing anti ...19999987159
a novel cellular protein, p60, interacting with both herpes simplex virus 1 regulatory proteins icp22 and icp0 is modified in a cell-type-specific manner and is recruited to the nucleus after infection.herpes simplex virus 1 encodes two multifunctional regulatory proteins, infected-cell proteins 22 and 0 (icp22 and icp0). icp0 is a promiscuous transactivator, whereas icp22 is required in vivo and for efficient replication and expression of a subset of late (gamma2) genes in rodent or rabbit cell lines and in primary human cell strains (restrictive cells) but not in hep-2 or vero (permissive) cells. we report the identification in the yeast two-hybrid system of a cellular protein designated p60 ...199910196275
icp22 and the ul13 protein kinase are both required for herpes simplex virus-induced modification of the large subunit of rna polymerase ii.herpes simplex virus type 1 (hsv-1) infection alters the phosphorylation of the large subunit of rna polymerase ii (rnap ii), resulting in the depletion of the hypophosphorylated and hyperphosphorylated forms of this polypeptide (known as iia and iio, respectively) and induction of a novel, alternatively phosphorylated form (designated iii). we previously showed that the hsv-1 immediate-early protein icp22 is involved in this phenomenon, since induction of iii and depletion of iia are deficient ...199910364308
equine herpesvirus 1 gene 12 can substitute for vmw65 in the growth of herpes simplex virus (hsv) type 1, allowing the generation of optimized cell lines for the propagation of hsv vectors with multiple immediate-early gene defects.herpes simplex virus (hsv) has often been suggested for development as a vector, particularly for the nervous system. considerable evidence has shown that for use of hsv as a vector, immediate-early (ie) gene expression must be minimized or abolished, otherwise such vectors are likely to be highly cytotoxic. mutations of vmw65 which abolish ie promoter transactivating activity may also be included to reduce ie gene expression generally. however, when vmw65 mutations are combined with an ie gene ...199910438830
comparison of the pathogenesis of acute equine herpesvirus 1 (ehv-1) infection in the horse and the mouse model: a review.the mouse models of the respiratory and abortion forms of equine herpesvirus 1 (ehv-1) infection have been used to investigate the vaccine potential of various ehv-1 immunogens, the effect of antiviral agents on ehv-1 infection and the pathogenicity of ehv-1 strain variants and deletion or insertional mutants. this review examines the similarities and differences in the pathogenesis of primary ehv-1 infection in the natural host, the horse, and in the mouse by comparing tissue tropism, clinical ...199910501157
epidemiological studies of equine herpesvirus 1 (ehv-1) in thoroughbred foals: a review of studies conducted in the hunter valley of new south wales between 1995 and 1997.sero-epidemiological studies conducted between 1995 and 1997 on two large thoroughbred stud farms in the hunter valley of nsw showed clear evidence of ehv-1 infection in foals as young as 30 days of age. similarly, serological evidence suggested that these foals were infected with ehv-1 from their dams or from other lactating mares in the group, with subsequent foal to foal spread of infection prior to weaning. these studies also provided evidence of ehv-1 infection of foals at and subsequent to ...199910501158
potential of dna-mediated vaccination for equine herpesvirus 1.the potential of dna-mediated immunisation to protect against equine herpesvirus 1 (ehv-1) disease was assessed in a murine model of ehv-1 respiratory infection. intramuscular injection with dna encoding the ehv-1 envelope glycoprotein d (gd) in a mammalian expression vector induced a specific antibody response detectable by two weeks and maintained through 23 weeks post injection. immune responses were proportional to the dose of dna and a second injection markedly enhanced the antibody respons ...199910501160
characteristics of glycoprotein b of equine herpesvirus 1 expressed by a recombinant baculovirus.a recombinant baculovirus (bac-egb) containing the complete open reading frame of equine herpesvirus 1 glycoprotein b (ehv-1 gb) expressed recombinant products of 107-133 kda, 58-75 kda and 53-57 kda, corresponding to ehv-1 gb precursor, large and small subunits respectively. high molecular mass products (>200 kda) in the bac-egb infected insect cells were consistent with oligomerisation of the recombinant ehv-1 gb products, and analysis with tunicamycin and endoglycosidases indicated that the b ...199910501161
colocalization of the herpes simplex virus 1 ul4 protein with infected cell protein 22 in small, dense nuclear structures formed prior to onset of dna synthesis.herpes simplex virus 1 infected cell protein 22 (icp22) localizes in small, dense nuclear bodies of primate cells early in infection and in the more diffuse replicative complexes after the onset of dna synthesis. ul4, a gamma2 protein, localizes in cytoplasm and in the small nuclear structures containing icp22 but not in replicative complexes. in rabbit skin cells, both icp22 and ul4 localize in the dense nuclear bodies late in infection. the results suggest that the small nuclear structures per ...199910233976
capsid structure of simian cytomegalovirus from cryoelectron microscopy: evidence for tegument attachment sites.we have used cryoelectron microscopy and image reconstruction to study b-capsids recovered from both the nuclear and the cytoplasmic fractions of cells infected with simian cytomegalovirus (scmv). scmv, a representative betaherpesvirus, could thus be compared with the previously described b-capsids of the alphaherpesviruses, herpes simplex virus type 1 (hsv-1) and equine herpesvirus 1 (ehv-1), and of channel catfish virus, an evolutionarily remote herpesvirus. nuclear b-capsid architecture is ge ...19999971801
non-essential loci in the bamhi-i and -f fragments of the hvt fc126 genome.the sequence of bamhi-i fragment of the herpesvirus of turkeys (hvt) fc126 strain dna was analyzed for the presence of potential open reading frames (orfs). four complete (orfs 2 to 5) and 2 partial orfs (orfs 1 and 6) were detected. orfs 2 and 3 were homologous to the hsv-1 ul55 and the ehv-1 gene 3, respectively. the orf 6 was already partially sequenced by smith et al. (virology 207, 205-216, 1995), and was homologous to a marek's disease virus (mdv) orf located in a similar position (orf 21; ...199910696442
characterization of the transactivation domain of the equine herpesvirus type 1 immediate-early protein.equine herpesvirus type 1 (ehv-1) possesses a sole diploid immediate early gene (ie) that encodes a major regulatory protein of 1487 amino acids capable of modulating gene expression from both early and late promoters and also of trans-repressing its own promoter. using a series of gal-4-ie fusion constructs, we previously demonstrated that the minimal transactivation domain (tad) of the ie protein maps within amino acids 3-89. additional studies revealed that that the carboxyl terminus of the i ...199910581386
functional anatomy of herpes simplex virus 1 overlapping genes encoding infected-cell protein 22 and us1.5 protein.earlier studies have shown that (i) the coding domain of the alpha22 gene encodes two proteins, the 420-amino-acid infected-cell protein 22 (icp22) and a protein, us1.5, which is initiated from methionine 147 of icp22 and which is colinear with the remaining portion of that protein; (ii) posttranslational processing of icp22 mediated largely by the viral protein kinase ul13 yields several isoforms differing in electrophoretic mobility; and (iii) mutants lacking the carboxyl-terminal half of the ...199910196329
gene arrangement and rna transcription of the bamhi fragments k and m2 within the non-oncogenic marek's disease virus serotype 2 unique long genome region.we determined the nucleotide sequence of a 6593 bp fragment of the marek's disease virus serotype 2 (mdv2) unique long region located in the right part of genomic bamhi-m2 and the adjacent part of bamhi-k fragments. within this region five complete open reading frames (orfs) were identified whose deduced amino acid sequences exhibited homology to the ul53 (glycoprotein k), ul54 (immediate early regulatory protein icp27), and ul55 gene products of herpes simplex virus type 1 (hsv-1). homologue to ...199910225279
detection of equine herpesvirus types 2 and 5 (ehv-2 and ehv-5) in przewalski's wild blood samples of seven captive equid species from four german zoos ehv-1 specific antibodies were detected in 76% and ehv-4 specific antibodies in 73% of the 55 animals, whereas 93% were tested positive for ehv-2 and ehv-5, respectively. in only one blood sample from a przewalski's wild horse ehv-4 dna was amplified by pcr. from seven przewalski's wild horses ehv-2, and from another one ehv-5 was isolated by cocultivation. the identity of the virus isolates was verified by pcr and restriction ...199910365167
[rhinopneumonia or mycotoxin intoxication? neurologic phenomena in horses from a riding school].in the course of several days most of the 40 riding-school horses turned out in paddocks developed ataxia of variable severity. five of these horses showed severe ataxia and tremors, became paralyzed and were euthanized. eleven privately-owned horses which were stabled on the same premises showed no clinical signs. the most likely diagnosis seemed to be the 'neurological form of ehv1', although the signs were not entirely typical. a few weeks later a second outbreak occurred among the riding-sch ...199910596400
infection with equine herpesvirus 1 (ehv-1) strain hvs25a in pregnant mice.the abortigenic effects of equine herpesvirus 1 (ehv-1) strain hvs25a, given intranasally, were assessed in pregnant balb/c, c57bl/6j and quakenbush mice at day 16 of pregnancy. all ehv-1-infected balb/c mice showed clinical signs typical of ehv-1-induced disease, together with evidence of abortion. however, although there were fetal and neonatal deaths in some c57bl/6j and quakenbush litters, the respiratory and systemic effects of ehv-1 infection in the dams were inconsistent. balb/c dams were ...199910098013
truncation of the c-terminal acidic transcriptional activation domain of herpes simplex virus vp16 renders expression of the immediate-early genes almost entirely dependent on icp0.the herpes simplex virus (hsv) proteins vp16 and icp0 play key roles in stimulating the onset of the viral lytic cycle. we sought to explore the regulatory links between these proteins by studying the phenotypes of viral mutants in which the activation functions of both were simultaneously inactivated. this analysis unexpectedly revealed that truncation of the c-terminal transcriptional activation domain of vp16 (allele v422) in an icp0-deficient background almost completely eliminated immediate ...199910559282
characterisation of ie and ul5 gene products of equine herpesvirus 1 using dna inoculation of mice.the equine herpesvirus 1 (ehv-1) strain hvs25a regulatory genes ie and ul5, encoding homologues of herpes simplex virus 1 (hsv-1) icp4 and icp27 respectively, were cloned into a eukaryotic expression vector and the dna injected intramuscularly into mice. antibodies produced in this way detected the ie or ul5 gene products as diffuse material in nuclei of rk13 cells transfected with the individual genes but as discrete punctate or large aggregates in rk13 cells infected with ehv-1. western blotti ...200011205113
application of a type-specific enzyme-linked immunosorbent assay for equine herpesvirus types 1 and 4 (ehv-1 and -4) to horse populations inoculated with inactivated ehv-1 vaccine.a type-specific enzyme-linked immunosorbent assay (elisa) using equine herpesvirus types 1 (ehv-1) and 4 (ehv-4) glycoprotein g was applied for sero-epizootiology of ehv infections in japan. recently, an inactivated ehv-1 vaccine has been administered to racehorses for prevention of upper respiratory disease. to examine the effect of the vaccination on the result of the elisa, 6 horses were experimentally inoculated three times intramuscularly or intranasally with inactivated ehv-1 vaccine. sera ...200010945284
use of apathogenic vaccinia virus mva expressing ehv-1 gc as basis of a combined recombinant mva/dna vaccination scheme.the nonreplicating chicken adapted vaccinia virus strain mva was used in a combined vaccine scheme. using the equine herpesvirus type 1 (ehv-1) encoded complement-receptor glycoprotein c as antigen, only poor antibody response was induced by exclusive vaccination with dna plasmids. the administration of recombinant mva followed by plasmid immunization elicited both humoral and cellular immune responses in hamster comparable to ehv-1 full virus vaccines. our results indicate that recombinant cons ...200010618528
a prime-boost immunization strategy with dna and recombinant baculovirus-expressed protein enhances protective immunogenicity of glycoprotein d of equine herpesvirus 1 in naïve and infection-primed mice.the immunogenicity and protective efficacy afforded by intramuscular inoculation of plasmid dna encoding equine herpesvirus 1 (ehv-1) glycoprotein d (gd) followed by ehv-1 gd expressed by a recombinant baculovirus was assessed in a murine model of ehv-1 respiratory infection. compared with mice inoculated with dna or protein only, mice inoculated with the combination of gd dna and protein had enhanced elisa and neutralizing antibody titres to ehv-1 and had accelerated clearance of virus from lun ...200010618534
the ends on herpesvirus dna replicative concatemers contain pac2 cis cleavage/packaging elements and their formation is controlled by terminal cis sequences.herpesviruses have large double-stranded linear dna genomes that are formed by site-specific cleavage from complex concatemeric intermediates. in this process, only one of the two genomic ends are formed on the concatemer. although the mechanism underlying this asymmetry is not known, one explanation is that single genomes are cleaved off of concatemer ends in a preferred direction. this implies that cis elements control the direction of packaging. two highly conserved cis elements named pac1 an ...200010627574
the disappearance of cyclins a and b and the increase in activity of the g(2)/m-phase cellular kinase cdc2 in herpes simplex virus 1-infected cells require expression of the alpha22/u(s)1.5 and u(l)13 viral uninfected cells the g(2)/m transition is regulated by cyclin kinase complex containing cdc2 and, initially, cyclin a, followed by cyclin b. cdc2 is downregulated through phosphorylation by wee-1 and myt-1 and upregulated by cdc-25c phosphatase. we have examined the accumulation and activities of these proteins in cells infected with wild type and mutants of herpes simplex virus 1. the results were as follows. (i) cyclin a and b levels were reduced beginning 4 h after infection and were undet ...200010590085
shivering in a thoroughbred 11-year-old mare presented with neuromuscular deficits and what resembled shivering in the left hind limb. on necropsy, there was no evidence of denervation atrophy of the left hind gastrocnemius muscle. the spinal cord had a small, right-sided lesion at c3-c4 and c4-c5. tests for equine herpesvirus-1 and sarcocystis spp. were negative.200010723600
replication of equine herpesvirus type 1 in freshly isolated equine peripheral blood mononuclear cells and changes in susceptibility following mitogen the present study, the outcome of an inoculation of equine peripheral blood mononuclear cells (pbmc) with equine herpesvirus type 1 (ehv-1) was studied in vitro. cytoplasmic and plasma membrane expression of viral antigens, intra- and extracellular virus titres, and plaque formation in co-culture were determined. ehv-1 replicated in monocytes, although in a highly restricted way. viral antigens were found at maximum levels (8.7% of the monocytes) at 12 h post-infection. the infection was prod ...200010640538
development and validation of a monoclonal antibody blocking elisa for the detection of antibodies against both equine herpesvirus type 1 (ehv1) and equine herpesvirus type 4 (ehv4).a monoclonal antibody blocking elisa was developed for the detection of antibodies directed against either ehv1 or ehv4. for this purpose, we selected a monoclonal antibody directed against a cross-reactive, conservative and immunodominant epitope of both ehv1 and ehv4. high antibody titres were found in rabbit antisera and spf-foal antisera infected with either ehv1 or ehv4. after experimental challenge of conventional horses with ehv1 or ehv4 significant increases in cf and elisa titres were f ...200010665532
[relevance of infection with equine herpesvirus 1 (ehv-1) in a german thoroughbred stud: vaccination, abortion and diagnosis].the aim of the present study was to clarify whether an ehv-1 induced abortion can be prognosticated by an increase of antibody titres, virus shedding and/or viraemia and whether the current abortion diagnostic is suitable. in this context the immune response post immunization and a possible reactivation were of great interest. for this purpose blood samples of 32 mares between the ages of 5-21 years were regularly investigated during a period of two years before and after vaccination and pregnan ...200010726362
equine immunity to viruses.the identification of some of the adaptive immune responses to infection with equine viruses has been the first step toward rational immunoprophylactic design. sufficient knowledge of infection-induced immunity and informed estimates of the requirements for long-term immunity for eiv have now been obtained. thus, the future for inactivated eiv vaccines is promising now that new adjuvants have been applied to induce cellular immunity and safe methods have been designed to stimulate virus-neutrali ...200010752138
ataxia and paresis with equine herpesvirus type 1 infection in a herd of riding school outbreak of neurologic disease associated with serologic evidence of equine herpesvirus type 1 (ehv-1) infection occurred in a herd of 46 riding school horses. ataxia and paresis were observed in 14 geldings and 5 barren mares. eight affected horses had distal limb edema, 1 horse had a head tilt, and 3 others had urinary incontinence. other clinical signs included fever, depression, and inappetance in 30 horses. seven horses with neurologic signs were treated with acyclovir. serum neutralizin ...200010772493
an equine herpesvirus 1 (ehv1) abortion storm at a riding outbreak of ehv1 abortions occurred at a riding school in the netherlands in 1991. seven of twelve pregnant mares aborted, and another foal died at 8 days of age. six abortions occurred within 12 days in march after an initial abortion on 8 february. four mares delivered live foals. virological examination of four aborted foals revealed an ehv1 infection. serological results for paired sera from 17 horses suggested, that the initial abortion on 8 february was the index case, and probably caus ...200010789515
differentiation and genomic and antigenic variation among fetal, respiratory, and neurological isolates from ehv1 and ehv4 infections in the netherlands.ten monoclonal antibodies (mabs) were produced against equine herpes virus type 1 (ehv1). two appeared type-specific, while the other eight were directed against epitopes common to both ehv1 and ehv4. two mabs directed against the glycoprotein gp2 recognized linear epitopes, as demonstrated by western blotting. with pools of type-specific mabs, 282 field isolates were typed in an immunoperoxidase monolayer assay (ipma). from a total of 254 fetal or neonatal isolates, 244 (96%) were typed as ehv1 ...200010789516
virulence of the v592 isolate of equid herpesvirus-1 in ponies.the v592 strain of equid herpesvirus-1 (ehv-1), which was originally isolated from a fetus during an abortion epizootic, has proved to be of low virulence in infection studies. five welsh mountain pony mares and one foal were challenged intranasally or by aerosol with this isolate, and monitored clinically and virologically. all six animals shed virus in nasopharyngeal mucus, and viraemia was recorded from day 7 post-infection (pi). pathological investigations revealed mild rhinitis and bronchio ...200010805982
incidence of equine herpesvirus 1 infection in thoroughbred weanlings on two stud farms. 200010840577
equid herpesvirus 1: platelets and alveolar macrophages are potential sources of activated tgf-b1 in the horse.cell mediated responses to equid herpesvirus 1 (ehv-1) are of short duration in vivo and require considerable expansion to be detected in vitro. raised serum levels of active transforming growth factor b (tgf-b1) have been shown to depress proliferative t cell responses in experimental infections with ehv-1 in ponies. the present work indicates that latent transforming growth factor b (tgf-b1) is present in circulating platelets, lymph node, bronchial epithelium and alveolar macrophages. activat ...200010889300
the catalytic subunit of herpes simplex virus type 1 dna polymerase contains a nuclear localization signal in the ul42-binding region.the herpes simplex virus type 1 dna polymerase consists of a catalytic subunit (pol or ul30) and a processivity factor (ul42). the pol/ul42 interaction, which occurs through the extreme c-terminus of pol, is essential for hsv-1 replication and thus represents a valid target for drug inhibition. we recently showed (a. loregian et al. (1999) proc. natl. acad. sci. usa 96, 5221-5226) that an oligopeptide corresponding to the 27 c-terminal amino acids of pol, when delivered into herpes simplex virus ...200010891416
utilisation of bacteriophage display libraries to identify peptide sequences recognised by equine herpesvirus type 1 specific equine sera.three filamentous phage random peptide display libraries were used in biopanning experiments with purified igg from the serum of a gnotobiotic foal infected with equine herpesvirus-1 (ehv-1) to enrich for epitopes binding to anti-ehv-1 antibodies. the sequences of the amino acids displayed were aligned with protein sequences of ehv-1, thereby identifying a number of potential antibody binding regions. presumptive epitopes were identified within the proteins encoded by genes 7 (dna helicase/prima ...200010921846
equine herpesvirus 1 (ehv-1) glycoprotein d dna inoculation in horses with pre-existing ehv-1/ehv-4 antibody.we have shown previously that equine herpesvirus 1 (ehv-1) glycoprotein d (gd) dna elicited protective immune responses against ehv-1 challenge in murine respiratory and abortion models of ehv-1 disease. in this study, 20 horses, all with pre-existing antibody to ehv-4 and two with pre-existing antibody to ehv-1, were inoculated intramuscularly with three doses each of 50, 200 or 500microg ehv-1 gd dna or with 500microg vector dna. in 8 of 15 horses, inoculation with ehv-1 gd dna led to elevated ...200010946142
immunization of balb/c mice with dna encoding equine herpesvirus 1 (ehv-1) glycoprotein d affords partial protection in a model of ehv-1-induced abortion.dna-mediated immunization was assessed in a murine model of equine herpesvirus 1 (ehv-1) abortion. whilst there are differences between the model and natural infection in the horse, literature suggests that ehv-1 infection of pregnant mice can be used to assess the potential ability of vaccine candidates to protect against abortion. female balb/c mice were inoculated twice, 4 weeks apart, with an expression vector encoding ehv-1 glycoprotein d (gd dna). they were mated 15 days after the second i ...200010973696
demonstration of equine herpesvirus-1 gene expression in the placental trophoblasts of naturally aborted equine fetuses.equine herpesvirus-1 (ehv-1) infection was demonstrated in the lung tissue of seven aborted fetuses by immunohistochemical labelling and polymerase chain reaction. the placentas of the fetuses were also examined by non-isotopic in-situ hybridization for the ehv-1 glycoprotein b (gb) gene. positive hybridization signals were observed in the cytoplasm of trophoblasts, especially in microcotyledons, of all seven placentas, and in villous epithelium of the allantochorion of six placentas. despite th ...200011032664
quantitation of virus-specific classes of antibodies following immunization of mice with attenuated equine herpesvirus 1 and viral glycoprotein d.the antibody responses of cba/j mice infected intranasally (i.n.) with either the attenuated kya strain or the pathogenic racl11 strain of equine herpesvirus 1 (ehv-1) or immunized with recombinant glycoprotein d (rgd) were investigated using the elispot assay to measure ehv-1-specific antibody-secreting cells (asc) in the regional lymphoid tissue of the respiratory tract. igg, iga, and igm asc specific for ehv-1 were detected in the mediastinal lymph nodes (mln) and lungs 2 weeks after i.n. inf ...200010704356
prevalence of equine herpesvirus type 1 latency detected by polymerase chain this study, an improved polymerase chain reaction (pcr) was used for detection of dna of latent ehv-1 strains from several sources. three pairs of oligonucleotide primers spanning fragments of 333 bp, 226 bp and 268 bp of the thymidine kinase (tk) gene, and one primer pair spanning 225 bp of the glycoprotein c (gc) gene were used in specific amplifications. primers for ehv-4 pcr were also designed. restriction digests with taqi confirmed the identity of tk pcr fragments from ehv-1. the sensit ...200011043940
severe murine lung immunopathology elicited by the pathogenic equine herpesvirus 1 strain racl11 correlates with early production of macrophage inflammatory proteins 1alpha, 1beta, and 2 and tumor necrosis factor alpha.the cba mouse model was used to investigate the immunopathology induced in the lung by the pathogenic equine herpesvirus 1 (ehv-1) strain racl11 in comparison to infection with the attenuated vaccine candidate strain kya. intranasal infection with kya resulted in almost no inflammatory infiltration in the lung. in contrast, infection with the pathogenic racl11 strain induced a severe alveolar and interstitial inflammation, consisting primarily of lymphocytes, macrophages, and neutrophils. infect ...200011024132
differences in determinants required for complex formation and transactivation in related vp16 proteins.vp16-h is an essential structural protein of herpes simplex virus type 1 (hsv-1) and is also a potent activator of virus immediate-early (ie) gene expression. current models of functional determinants within vp16-h indicate that it consists of two domains, an n-terminal domain involved in recruiting vp16-h to a multicomponent dna binding complex with two host proteins, oct-1 and host cell factor (hcf), and an acidic c-terminal domain exclusively involved in transactivation. vp16-e, from equine h ...200011024140
fatal nonneurological ehv-1 infection in a yearling filly.a case of fatal nonneurological equine herpesvirus 1 (ehv-1) infection in a yearling filly is described. gross lesions included extensive pulmonary edema, prominent laryngeal lymphoid follicles, and congestion and edema of the dorsal third ventricle choroid plexus. histologically, there was vasculitis, hemorrhage, and edema in the lungs and dorsal third ventricle choroid plexus as well as mild intestinal crypt necrosis with occasional intranuclear inclusion bodies. the perivascular and vascular ...200011105961
equine herpesvirus 1 (ehv-1) glycoprotein m: effect of deletions of transmembrane domains.equine herpesvirus 1 (ehv-1) recombinants that carry either a deletion of glycoprotein m (gm) or express mutant forms of gm were constructed. the recombinants were derived from strain kentucky a (kya), which also lacks genes encoding ge and gi. plaques on rk13 cells induced by the gm-negative kya were reduced in size by 80%, but plaque sizes were restored to wild-type levels on gm-expressing cells. electron microscopic studies revealed a massive defect in virus release after the deletion of gm i ...200011118370
efficacy of an inactivated vaccine for equine herpesvirus type 1 in a novel hamster model.a vaccine designated f.ehv1(s(-))bhk, which was prepared by formaldehyde treatment of equine herpesvirus type 1 (ehv1)-infected baby hamster kidney cells, stimulated neutralising antibodies in hamsters and rabbits and protected new-born hamsters against a lethal challenge with ehv1 by vaccination of their pregnant mothers. the preparative method was then modified to eliminate virus particles and intraparticulate dna, producing a vaccine designated bg.f.ehv1(s(-))bhk. this modification stimulated ...200010773735
a rapid and sensitive pcr-based diagnostic assay to detect bovine herpesvirus 1 in routine diagnostic submissions.we describe a rapid, sensitive and specific polymerase chain reaction (pcr) assay for the detection of bhv1 dna in a range of routine diagnostic submissions without the need for prior virus isolation. the assay, which is based on the selected amplification of a portion of the viral tk gene, detected both bhv1.1 and bhv1.2 subtypes in a panel of 15 characterised field isolates, and its sensitivity was estimated to be <0.125 tcid(50). bhv2, alcephaline herpesvirus, bhv4, equine herpesvirus 1 (ehv1 ...200010889405
ehv-1 glycoprotein d (ehv-1 gd) is required for virus entry and cell-cell fusion, and an ehv-1 gd deletion mutant induces a protective immune response in mice.insertional mutagenesis was used to construct an equine herpesvirus 1 (ehv-1) mutant in which the open reading frame for glycoprotein d was replaced by a lacz cassette. this gd deletion mutant (delta gd ehv-1) was unable to infect normally permissive rk cells in culture, but could be propagated in an ehv-1 gd-expressing cell line (rk/gd). phenotypically complemented delta gd ehv-1 was able to infect rk cells, but did not spread to form syncytial plaques as seen with wild type ehv-1 or with delta ...200011205124
solid matrix-antibody-antigen complexes incorporating equine herpesvirus 1 glycoproteins c and d elicit anti-viral immune responses in balb/c (h-2k(d)) and c3h (h-2k(k)) mice.glycoproteins c and d (gc and gd) derived from equine herpesvirus 1 (ehv-1)-infected cells were incorporated into individual solid matrix-antibody-antigen (smaa) complexes and administered to balb/c (h-2k(d)) and c3h (h-2k(k)) mice. antibodies against each of the glycoproteins were produced that neutralised virus infectivity and mediated the lysis of ehv-1-infected target cells in the presence of complement. immunoglobulin (ig)g2b was the predominant antibody isotype produced in balb/c mice agai ...200011115713
development of a differential multiplex pcr assay for equine herpesvirus 1 and 4 as a diagnostic this study, a multiplex polymerase chain reaction (pcr) procedure was developed for differentiation of strains and field isolates of equine herpesvirus type 1 (ehv-1) and type 4 (ehv-4). specific oli-gonucleotide primers were combined to amplify the thymidine kinase (tk) gene region of ehv-1 and ehv-4, which would yield fragments of different lengths for each virus in the same amplification reaction. the specificity of the largest pcr amplicon for ehv-4 was confirmed by restriction digestion ...200010900826
characterization of the trans-activation properties of equine herpesvirus 1 eicp0 protein.the eicp0 protein of equine herpesvirus 1 (ehv-1) is an early, viral regulatory protein that independently trans-activates ehv-1 immediate-early (ie), early, gamma1 late, and gamma2 late promoters. to assess whether this powerful trans-activator functions in conjunction with three other ehv-1 regulatory proteins to activate expression of the various classes of viral promoters, transient cotransfection assays were performed in which effector plasmids expressing the eicp22, eicp27, and ie proteins ...200010627530
the eicp22 protein of equine herpesvirus 1 physically interacts with the immediate-early protein and with itself to form dimers and higher-order complexes.the eicp22 protein (eicp22p) of equine herpesvirus 1 (ehv-1) is an early protein that functions synergistically with other ehv-1 regulatory proteins to transactivate the expression of early and late viral genes. we have previously identified eicp22p as an accessory regulatory protein that has the ability to enhance the transactivating properties and the sequence-specific dna-binding activity of the ehv-1 immediate-early protein (iep). in the present study, we identify eicp22p as a self-associati ...200010627553
expression of two related viral early genes in epstein-barr virus-associated tumors.the transcription of two early "leftwardly" expressed genes carrying repetitive sequences, ir2 and ir4, has been studied for epstein-barr virus-associated tumors, and for established b-cell lines, using sequence-specific probes generated for this purpose. whereas the ir4 transcript was identified in every tumor and cell line assessed (except b95-8, with a deletion that removes the gene), expression of the ir2 gene was restricted to b lymphocytes. though the promoters for both transcripts lie wit ...200010684296
pseudorabies virus glycoprotein m inhibits membrane fusion.a transient transfection-fusion assay was established to investigate membrane fusion mediated by pseudorabies virus (prv) glycoproteins. plasmids expressing prv glycoproteins under control of the immediate-early 1 promoter-enhancer of human cytomegalovirus were transfected into rabbit kidney cells, and the extent of cell fusion was quantitated 27 to 42 h after transfection. cotransfection of plasmids encoding prv glycoproteins b (gb), gd, gh, and gl resulted in formation of polykaryocytes, as ha ...200010888614
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