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[a new approach to dna synthesis from synthetic oligonucleotides. synthesis of dna coding for the repeated antigenic determinant of foot and mouth disease virus].a rapid method for assembly of dna from synthetic oligodeoxynucleotides has been developed which involves separate ligation of top- and bottom-strand oligonucleotides followed by filling in 3'-ends of the duplex formed, blunt end cloning into a specialized vector pbbv, and recovery of the synthetic dna from the recombinant plasmid by means of restriction nuclease bbvii. the method allows for many oligonucleotides to be ligated at once, with no intermediates being isolated, and any dna to be reco ...19872436628
antigenic comparison of the polypeptides of foot-and-mouth disease virus serotypes and other picornaviruses.the cross-reactivity of proteins coded for by the seven serotypes of foot-and-mouth disease virus (fmdv) was assessed by reaction of infected cell lysates with polyclonal and monospecific antisera against the structural and nonstructural proteins of fmdv type a12 strain 119ab. it was shown that the homologous polypeptides from most serotypes are antigenically related. the least cross-reactivity occurred between vp1, vp3, and the protease (3c) of type a12 and south african territories types 1 and ...19872437694
synthesis of fusion proteins with multiple copies of an antigenic determinant of foot-and-mouth disease virus.a series of four expression plasmids coding for fusion proteins containing foot-and-mouth disease virus (fmdv) sequences was constructed. the fusion proteins contain a large part of beta-galactosidase from escherichia coli preceded (n-terminal) by 1, 2, 4 or 8 repeats of the antigenic determinant of fmdv consisting of amino acids 137-162 of the capsid polypeptide vp1. all four fusion proteins were efficiently produced in e. coli host bacteria. immunization of rabbits resulted in fmdv-specific, n ...19862436976
reactivity with monoclonal antibodies of viruses from an episode of foot-and-mouth disease.a panel of 12 monoclonal antibodies (mabs) raised against foot-and-mouth disease virus (fmdv) of serotype c1 (fmdv c-s8c1) and 11 mabs raised against other fmdvs have been used to evaluate the reactivity of 14 isolates of fmdv of serotype c1 (series fmdv c-s), 12 of them from one disease episode (spain 1979-1982). the assays used were immunoelectrotransfer blot, immunodot and neutralization of infectivity. none of the isolates could be clearly distinguished by its reactivity with 6 non-neutraliz ...19872446442
non-responsiveness to a foot-and-mouth disease virus peptide overcome by addition of foreign helper t-cell determinants.study of the immune response to synthetic antigens has shown that uncoupled peptides can realize their potential as vaccines only if they contain domains that react with helper t-cell receptors and ia antigens in addition to antibody binding sites. here we consider whether genetically restricted non-responsiveness to an uncoupled peptide could be overcome by synthesizing a peptide with an additional helper t-cell epitope from a different protein. we demonstrate that h-2d mice, which are non-resp ...19872444892
improved immunogenicity of a peptide epitope after fusion to hepatitis b core protein.synthetic vaccines for viral diseases can use defined regions of viral proteins as immunogens: the peptide sequence of amino acids 141-160 of the vp1 protein of foot and mouth disease virus (fmdv) elicits virus-neutralizing antibodies to protect guinea pigs, cattle and pigs either when coupled to a carrier protein or when administered in liposomes or in incomplete freund's adjuvant. the immune response to these peptides is much lower than that to complete virus particles and the same sequence fu ...19872446137
[synthetic peptides simulating the protective epitopes of vp1 protein of foot-and-mouth disease virus type o and a].in a search of novel approaches to cattle protection from foot-and-mouth disease we have prepared a series of peptides from the major antigenic region 130-160 of the vp1 protein. the 144-159 peptide as well as 141-152, 141-148, 148-159 segments (strain o1k) were inactive in all in vitro and in vivo experiments on virus inhibiting. on the other band, synthetic 136-152, 136-148 o1k sequences as well as 131-149, 140-149 a22 sequences afforded 50 to 100% protection, both in the free state and conjug ...19872445357
fusion proteins with multiple copies of the major antigenic determinant of foot-and-mouth disease virus protect both the natural host and laboratory animals.proteins consisting of one, two or four copies of the amino acid sequence 137 to 162, which contains the major immunogenic site of vp1 of foot-and-mouth disease virus, attached to the n-terminus of beta-galactosidase have been expressed in escherichia coli cells. in guinea-pigs the protein containing one copy (p71) of the viral determinant elicited only low levels of neutralizing antibody whereas protective levels were elicited by the proteins containing two (p72) or four (p74) copies of the det ...19872447225
use of outer membrane protein phoe as a carrier for the transport of a foreign antigenic determinant to the cell surface of escherichia coli k-12.phoe protein is an abundant transmembrane protein from the escherichia coli k-12 outer membrane. a synthetic oligodeoxynucleotide corresponding to an antigenic determinant of the c-terminal part of the vp1 protein of foot-and-mouth disease virus was inserted into the phoe gene in an area corresponding to a cell surface-exposed region of the phoe protein. the level of expression of the hybrid protein was normal and the protein was incorporated into the outer membrane. the vp1-epitope was exposed ...19872449378
studies on the infectivity of foot-and-mouth disease virus rna using microinjection.foot-and-mouth disease virus (fmdv) rna, isolated as virion rna from purified virus particles or as total rna from infected cells, has been microinjected into nuclei and cytoplasms of bhk cells. when injected directly into the nucleus fmdv rna was not infectious, whereas cytoplasmic injection resulted in a high proportion of productive infections. infectivity microinjection assays on dilution series of various fmdv rnas showed that both single-stranded positive sense 35s rna and double-stranded ...19882448416
antigenic sites on foot-and-mouth disease virus type a10.a set of monoclonal antibodies was used to isolate nonneutralizable foot-and-mouth disease virus variants, and the rnas of the variants were sequenced. cross-neutralization studies and mapping of the amino acid changes indicated two major antigenic sites. the first site was trypsin sensitive and included the vp1 140 to 160 sequence. the second site was trypsin insensitive and included mainly vp3 residues. two minor sites were located near vp1 169 and on the c terminus of vp1. comparison with pol ...19882455819
genetic and immunogenic variations among closely related isolates of foot-and-mouth disease virus.genetic heterogeneity among closely related isolates of foot-and-mouth disease virus (fmdv) has been measured by direct sequencing of the vp1-coding-region rna for three new fmdvs of serotype c1 and by additional sequences of rna from previously reported isolates, all belonging to a single episode of disease [sobrino et al., gene 50 (1986) 149-159]. in the ten viruses compared, eight different vp1 are represented. the changes include amino acid substitutions at a critical antigenic determinant o ...19882453395
immunization against foot-and-mouth disease with synthetic peptides representing the c-terminal region of vp1.foot-and-mouth disease virus challenge experiments in guinea-pigs and immunoassays with a range of peptides equivalent to either or both of the sequences 141 to 158 and 200 to 213 of vp1 showed the most effective structure, in terms of protection, to be one in which both 'sites' were present with a minimum of additional amino acids. an 80 residue peptide comprising amino acids 134 to 213 was considerably less effective than 40 or 45 residue peptides. the major site for the induction of protectio ...19882457649
class i mhc-restricted cytotoxic t cells efficiently recognize haemagglutinin that is defective in protein folding and cell surface expressions.cytotoxic t-cell recognition of an engineered variant of the influenza viral haemagglutinin (ha), expressed in vaccinia virus, was investigated. we show that the insertion of a foot-and-mouth disease virus (fmdv) immunogenic peptide into the ha results in major disruption of its higher order structure with intracellular rather than cell surface localization accompanying the loss of conformational epitopes detected by antibody. in contradistinction to antibody, recognition of the chimaeric molecu ...19882467191
extensive antigenic heterogeneity of foot-and-mouth disease virus of serotype c.the antigenic behavior of 46 field isolates of foot-and-mouth disease virus (fmdv) of serotype c has been studied with a panel of 24 monoclonal antibodies (mabs) prepared against fmdv c1 or fmdv c3 indaial. reactivities were assayed by immunodot, immunoelectrotransfer blot, and neutralization of infectivity. the epitopes recognized by the 10 nonneutralizing mabs are conserved in all isolates analyzed. in contrast, extreme antigenic heterogeneity is documented with regard to reactivity with 14 ma ...19882460992
evidence for more than one important, neutralizing site on foot-and-mouth disease virus. brief report.using polyclonal sera raised against foot-and-mouth disease virus in susceptible animals, evidence was obtained for the existence of at least one further important antigenic site in addition to the neutralizing site on vp1 140-160.19882453185
novel low-molecular-weight synthetic vaccine against foot-and-mouth disease containing a potent b-cell and macrophage activator.most synthetic peptide vaccines described to date are effective only in combination with proteins and freund's adjuvant. the work describes a novel completely synthetic virus peptide vaccine, which consists of a synthetic activator of b cells and macrophages, covalently linked to an amphiphilic alpha-helical t-cell epitope. the low-molecular-weight vaccine of 3.4 kda developed against foot-and-mouth disease virus (fmdv) is composed of a synthetic vp1 (135-154) with a sequence homologous to an fm ...19892470215
molecular dynamics of the alpha-helical epitope of a novel synthetic lipopeptide foot-and-mouth disease virus vaccine.a novel synthetic foot-and-mouth disease virus (fmdv) peptide vaccine consisting of a synthetic b-cell and macrophage activator covalently linked to an amphiphilic alpha-helical t-cell epitope was developed. the low molecular weight vaccine of 3400 daltons is composed of virus vp1 antigenic determinant and the immunologically active lipotripeptide tripalmitoyl-s-glyceryl-cysteinyl-seryl-serine (p3css) as built-in adjuvant. the vaccine, tripalmitoyl-s-glyceryl-cysteinyl-seryl-seryl-fmdv-vp1 (vp1 ...19892470437
neutralizing epitopes of type o foot-and-mouth disease virus. i. identification and characterization of three functionally independent, conformational sites.eleven neutralizing monoclonal antibodies (mabs) were produced to the o1bfs 1860/67 strain of foot-and-mouth disease virus (fmdv), and were characterized for their ability to bind viral and subviral antigens in different elisa tests and to neutralize heterologous type o isolates. neutralization escape variants of the homologous virus, isolated under pressure from five of these mabs, were used in cross-neutralization tests with all of the 11 antibodies. these studies identified three functionally ...19892471811
neutralizing epitopes of type o foot-and-mouth disease virus. ii. mapping three conformational sites with synthetic peptide reagents.four neutralizing monoclonal antibodies (mabs), recognizing three functionally independent, conformational sites on type o foot-and-mouth disease virus (fmdv) failed to react with immobilized structural proteins or synthetic peptides but bound to the isolated capsid protein vp1 and peptides in solution. inhibition elisa techniques were, therefore, applied using peptide antigens and anti-peptide sera to block mab binding to virus particles, permitting the identification of those portions of the v ...19892471812
epitope mapping of foot-and-mouth disease virus with neutralizing monoclonal antibodies.epitopes of strain a22 iraq 24/64 of foot-and-mouth disease virus have been mapped with monoclonal antibodies (mabs). three methods were used: (i) an indirect elisa using an overlapping set of peptides, (ii) production of neutralization escape variants against each mab and (iii) sequencing of neutralization escape variants. the study has shown that the virus has at least three overlapping liner neutralizing epitopes within a major antigenic site on vp1. the presence of a second, conformational s ...19892471783
orientation of epitopes influences the immunogenicity of synthetic peptide dimers.the immunogenicity of synthetic peptide dimers based on epitope sequences derived from the mycobacterial 65-kda antigen and the foot and mouth disease virus (fmdv) vp1 protein was examined in inbred mice. the analysis was directed towards the potential helper role of a t cell stimulatory mycobacterial epitope (65-85) with respect to poorly immunogenic sites either from the same molecule (422-436) or from vp1 (141-160). the 65-85 repeat homodimer induced an antibody response in cba/ca but not in ...19882464496
host cell selection of antigenic variants of foot-and-mouth disease virus.foot-and-mouth disease virus (fmdv) a22 iraq 24/64 adapted to grow in bhk monolayer cells induced antibodies which neutralized many isolates belonging to the a serotype. plaque-purified virus isolated from this stock also induced broadly reactive antibodies, showing that this property is not due to the combined response to a mixture of variants in the original stock virus. however, viruses obtained by passage in suspension bhk cells of either the monolayer cell-adapted virus or a virus cloned fr ...19892471782
evidence for at least four antigenic sites on type o foot-and-mouth disease virus involved in neutralization; identification by single and multiple site monoclonal antibody-resistant mutants.neutralizing monoclonal antibodies raised against type o foot-and-mouth disease virus have been characterized on the basis of their reactivity with a panel of single site monoclonal antibody-resistant mutants which had defined three antigenic sites. five antibodies neutralized all these mutants, but by selecting further single site mutants with one of these antibodies it was possible to define a fourth site involved in virus neutralization. two monoclonal antibodies still neutralized these mutan ...19892471793
analysis of neutralizing antigenic sites on the surface of type a12 foot-and-mouth disease virus.a series of seven neutralizing monoclonal antibodies (nmabs) directed against type a12 foot-and-mouth disease virus was used to generate neutralization-resistant variants. both plaque reduction neutralization and microneutralization assays showed that the variants were no longer neutralized by the nmabs used to generate them, although some of the variants still reacted with the nmabs at high antibody concentrations. results of cross-neutralization studies by both plaque reduction neutralization ...19892467993
implications of a quasispecies genome structure: effect of frequent, naturally occurring amino acid substitutions on the antigenicity of foot-and-mouth disease virus.we provide evidence that the quasispecies nature (extreme genetic heterogeneity) of foot-and-mouth disease virus is relevant to the virus evading an immune response. a monoclonal antibody neutralizing the viral infectivity (clone sd6) recognizes an epitope located around a highly conserved sequence (amino acid sequence arg-gly-asp-leu-ala at positions 141-145) in the capsid protein vp1 of foot-and-mouth disease virus of serotype c1. the amino acid substitutions ala-138----thr and leu-147----ile ...19892474821
helper t-cell determinants in vaccine design.mice belonging to the h-2d haplotype do not respond to the 141-160 peptide of foot-and-mouth disease virus protein vp1. however, when defined 'foreign' helper t-cell determinants from ovalbumin or sperm whale myoglobin are added they do respond. these observations are likely to have important implications for the design of peptide vaccines.19892476143
a possible homology between immunodeficiency virus p24 core protein and picornaviral vp2 coat protein: prediction of hiv p24 antigenic sites.with the use of a sensitive sequence comparison algorithm, a homology has been suggested between the primary structures of simian immunodeficiency virus (siv) p24 core protein and foot-and-mouth disease virus (fmd) vp2 coat protein. since the fmd sequence is homologous to picornaviral vp2 sequences with known three-dimensional architecture and since the siv p24 sequence can be convincingly aligned with that from human immunodeficiency virus (hiv), it was possible to predict an eight-stranded bet ...19892498082
[antigenic structure of the foot-and-mouth disease virus. iii. immunogenic properties of synthetic peptides of the sequence of the immunodominant region of vp1 proteins of the o1k and a22 strains of foot-and-mouth virus].immunogenic and protective properties of uncoupled and klh-conjugated peptides covering the sequence of the immunodominant region of vp1 proteins of the o1k and a22 strains of foot-and-mouth disease virus have been studied. the uncoupled peptides 136-148 o1k, 136-152 o1k, 131-149 a22 and 140-149 a22 were shown to be immunogenic in guinea pigs and induced 50-100% protection against homologous virus. on the other hand, the a22 specific peptides, in contrast to the o1k peptides, were not immunogeni ...19892480134
selection of antigenic variants of foot-and-mouth disease virus in the absence of antibodies, as revealed by an in situ assay.antigenic variants of foot-and-mouth disease virus (fmdv) of serotype c (isolate c-s8c1) were selected upon serial passage of the virus in cell culture in the absence of anti-fmdv antibodies. the variants rose from frequencies of less than 10(-2) in the initial plaque-purified fmdv c-s8c1 preparation, to 0.1 to 1 in three passaged populations. the proportion of antigenic variants was quantified using a new in situ plaque immunotest. a nitrocellulose filter is applied to the agar overlay of a fmd ...19892481712
antigenic comparison of different foot-and-mouth disease virus types using monoclonal antibodies defining multiple neutralizing epitopes on fmdv a5 subtypes.thirteen monoclonal antibodies (mabs) were elicited with a5 spain-86 virus, the cause of the most recent foot-and-mouth disease virus (fmdv) outbreak in spain. the mabs were tested for ability to bind 140s virions and 12s protein subunits by liquid-phase radioimmunoassay (ria), and to bind vp1 capsid protein by western immunoblot assay. one of the thirteen mab was virion (140s) specific, seven recognized 140s and 12s subunits, one bound to 140s, 12s and vp1 and four were 12s specific. these mabs ...19892473578
[recombinant plasmids containing hybrid protein genes with antigenic determinants of the foot and mouth virus].plasmids have been constructed which contain genes coding for fused proteins including beta-galactosidase or human leukocyte interferon alpha 2 and monomeric or pentameric form of the main antigenic determinant of the foot-and-mouth disease virus (fmdv) serotype 01k. expression of the hybrid genes has been studied. it is shown that fused proteins, containing beta-galactosidase and the antigenic determinant (monomer or pentamer), interact specifically with anti-fmdv anti-sera and with antibodies ...19892473757
identification of virus neutralizing epitopes on naturally occurring variants of type a12 foot-and-mouth disease virus.four naturally occurring antigenic variants of foot-and-mouth disease virus type a12 were examined for their capacity to be neutralized by a number of monoclonal antibodies (mab) which recognize different sites on the virus surface. the vp1 coding region of the rna genome was sequenced and amino acid changes were determined for the variants. one of the neutralizing sites accounted for the differing antigenic properties of the variants and the epitope was mapped to amino acid residues 150-156 of ...19892483013
protective and immunostimulating activity of a low dose of cyclophosphamide in the experimental infection of mice with foot-and-mouth disease virus.administration to mice of a low, non-immunosuppressive dose of cyclophosphamide 4 days before infection with foot-and-mouth disease virus decreases viral replication, enhances the immune response against the virus and prevents pancreatic damage.19892536333
the three-dimensional structure of foot-and-mouth disease virus at 2.9 a resolution.the structure of foot-and-mouth disease virus has been determined at close to atomic resolution by x-ray diffraction without experimental phase information. the virus shows similarities with other picornaviruses but also several unique features. the canyon or pit found in other picornaviruses is absent; this has important implications for cell attachment. the most immunogenic portion of the capsid, which acts as a potent peptide vaccine, forms a disordered protrusion on the virus surface.19892537470
the suitability of different microtitre plates for detection of antibody to virus antigens by indirect elisa.to optimize enzyme linked immunosorbent assays (elisas) for the detection of virus-specific antibodies, a range of commercially available microtitre plates was evaluated for their ability to bind virus antigen. rinderpest virus and foot-and-mouth disease virus were investigated as target antigens. binding capacity for antigen, binding ratios (attachment of specific antibody versus that of non-immune antibody) and the variation in the results of the tests within and between plates were measured. ...19892541132
the nucleotide sequence of the structural-protein-coding region of foot-and-mouth disease virus serotype sat3.the nucleotide sequence coding for the structural proteins and nonstructural protein p2a has been determined for a foot-and-mouth disease virus (fmdv) isolated in africa. this virus, serotypically designated sat3 (south african territories type 3), shows about 60% homology at the nucleotide level to prototype viruses from the o, a and c serotypes of fmdv. the highest region of variability was shown in structural protein vp1, presumably a consequence of its position on the surface of the virus an ...19892541051
extensive cell heterogeneity during persistent infection with foot-and-mouth disease virus.coevolution of viruses and the host cells occurred in bhk-21 cell cultures persistently infected with foot-and-mouth disease virus (fmdv) (j. c. de la torre, e. martínez-salas, j. diez, a. villaverde, f. gebauer, e. rocha, m. dávila, and e. domingo, j. virol. 62:2050-2058, 1988). in the present report we provide evidence of an extreme phenotypic heterogeneity of the cells, which was generated in the course of persistence. a total of 248 stable cell clones isolated from fmdv carrier cultures at e ...19892535753
manipulation of antipeptide immune response by varying the coupling of the peptide with the carrier protein.antibodies were raised against synthetic peptides of two regions of the surface protein vp1 of foot-and-mouth disease virus. the peptides were conjugated with keyhole limpet hemocyanin via c- or n-terminal amino acid residues by use of different coupling agents. the fine specificity of the resulting antibodies was determined by pepscan methods. in general, amino acid residues specific for antibody recognition tended to be located opposite to those used for coupling with the carrier protein. depe ...19892538727
resistance to foot-and-mouth disease virus mediated by trans-acting cellular products.upon serial passage of bhk-21 cells persistently infected with foot-and-mouth disease virus (fmdv) c-s8c1, cells with increased resistance to the virus were selected (j. c. de la torre, e. martinez-salas, j. diez, and e. domingo, j. virol. 63:59-63, 1989). two highly resistant cell clones, 74a11 and 74d12, were transformed to puromycin resistance (purr) and were fused to bhk-21 cells transformed to neomycin resistance (neor). the hybrid neor purr cells showed the specific resistance to fmdv c-s8 ...19892539526
t cell-dependent induction of antibody against foot-and-mouth disease virus in a mouse model.nude and normal balb/c mice were primed by intravenous inoculation of purified, infectious foot-and-mouth disease virus (fmdv) type a24, strain cruzeiro. frequency estimation of antigen-specific antibody-secreting cells (asc) and thy 1+ t cells in the spleens of immunized mice identified that the igm response was similar for both nude and normal mice, whereas substantial numbers of both igg asc and thy 1+ cells were present in normal mice only. in contrast, nude and normal mouse sera both contai ...19892543745
serological prospects for peptide vaccines against foot-and-mouth disease virus.antibodies to a synthetic peptide corresponding to the 141 to 160 amino acid sequence of the protein vp1 of type o foot-and-mouth disease virus (fmdv) neutralize a wider range of type o isolates than anti-virion serum. extending this peptide at the amino terminus reduced the number of strains neutralized by the antipeptide sera. reactions with antisera to peptides representing non-contiguous native sequences showed that it was also possible to increase the number of strains effectively neutraliz ...19892479714
[primary structure of the a22 rna polymerase gene of the foot and mouth disease virus].complete nucleotide sequence of gene rna polymerase for the foot-and-mouth disease virus subtype a22 has been determined.19892545214
antibodies to foot-and-mouth disease virus infection associated (via) antigen: use of a bioengineered via protein as antigen in an elisa.an enzyme-linked immunosorbent assay (elisa) to detect antibodies to foot-and-mouth disease (fmd) virus infection associated (via) antigen (viral rna polymerase) in cattle sera, was developed using a bioengineered via (biovia) protein antigen. compared with the classical immunodiffusion test, with viral rna polymerase purified from infected cell cultures as antigen, this elisa was more sensitive. however, depending on the cattle population examined, sera with antibodies to viral rna polymerase, ...19892549685
evaluation of techniques to demonstrate foot-and-mouth disease virus in bovine tongue epithelium: comparison of the sensitivity of cattle, mice, primary cell cultures, cryopreserved cell cultures and established cell lines.tongue epithelia infected with each of the 7 serotypes of foot-and-mouth disease virus (fmdv) were used to evaluate in vivo and in vitro systems for the detection of fmdv. cattle inoculated by the intradermal route in the tongue (idl) and suckling mice inoculated intraperitoneally were compared for susceptibility to fmdv with freshly prepared bovine thyroid cell cultures; cultures from cryopreserved bovine thyroid, bone marrow, mammary gland, myocardium, tongue, ovary and kidney cells; cultures ...19892549683
modification of foot-and-mouth disease virus after serial passages in the presence of antiviral polyclonal sera.foot-and-mouth disease virus (fmdv) shows a remarkable antigenic variability. like other rna viruses, this virus has a high rate of mutation. it has been proposed that selection exerted by the host's antibodies could play a major role in the rapid evolution of fmdv. the present work reports the selection of fmdv antibody-resistant populations (nr), after serial passages of cloned fmdv a24 cruzeiro strain on secondary monolayers of bovine fetal kidney cells in the presence of subneutralizing anti ...19892548330
the cell attachment site on foot-and-mouth disease virus includes the amino acid sequence rgd (arginine-glycine-aspartic acid).the amino acid sequence rgd (arginine-glycine-aspartic acid) is highly conserved in the vp1 protein of foot-and-mouth disease virus (fmdv), despite being situated in the immunodominant hypervariable region between amino acids 135 and 160. rgd-containing proteins are known to be important in promoting cell attachment in several different systems, and we report here that synthetic peptides containing this sequence are able to inhibit attachment of the virus to baby hamster kidney (bhk) cells. inhi ...19892543752
sequences of capsid protein vp1 of two type a foot-and-mouth disease viruses.we have sequenced the nucleotides of the regions that encode the capsid protein vp1 of the foot-and-mouth disease viruses (fmdv) a5bernbeuren/1984 and a iran/1987. amino acid sequences and secondary protein structures are provided. both proteins consist of 212 amino acids. the sequences and secondary structures are compared to those of fmdv a22/cccp/64, a strain previously endemic in the near east. nucleotide divergency among the three sequences is highest for fmdv a5bernbeuren/1984 (18% compare ...19892548339
development of foot-and-mouth disease virus strain characterisation--a review. 19892552629
characterization of anti-idiotypic antibodies generated against foot-and-mouth disease virus neutralizing monoclonal antibodies.a series of seven neutralizing monoclonal antibodies (nmabs) against type a12 foot-and-mouth disease virus (fmdv) was used to induce polyclonal anti-idiotypic antibodies (anti-ids) in rabbits. the anti-ids were semi-purified through isotype affinity columns and assayed by solid-phase radioimmunoassay for cross-reactivity. nmabs which map to the same epitope on the virion appear to contain a common idiotype, and the corresponding anti-ids competitively inhibited the virus-nmab reaction. using a m ...19892550021
foot-and-mouth disease virus-neutralizing antibodies induced in mice by anti-idiotypic antibodies.a neutralizing monoclonal antibody (nmab) to foot-and-mouth disease virus (fmdv) was used as antibody-1 (ab1) to induce anti-idiotypic antibodies (a-idab) in rabbits. the rabbit a-idab (ab2) were isolated on protein a-sepharose, followed by cycles of separation on idiotype and isotype affinity columns. the specificity of the ab2 for the paratope of ab1 was determined by direct binding to ab1 in solid-phase radioimmunoassay (sp-ria), and by competition ria (c-ria) with virus for binding to the ab ...19892553588
specificity of enzyme-substrate interactions in foot-and-mouth disease virus polyprotein processing.a series of transcripts derived from fmdv cdna plasmids containing defined regions of the genome were translated in a rabbit reticulocyte lysate system. the products were analysed directly or following incubation with an fmdv-infected cell processing extract. processing by the l proteinase at the l/1a cleavage site occurred when most of the p1-2a protein was absent. substitution of sequences upstream of the 2c/3a cleavage site showed that the 3c proteinase was also able to cleave at an entirely ...19892554577
serological probes for some foot-and-mouth disease virus nonstructural proteins.foot-and-mouth disease virus (fmdv) o1 kaufbeuren-specific cdna fragments were subcloned into the e. coli expression vector prit.2t. fusion proteins thus produced in bacteria were purified by affinity chromatography and inoculated into rabbits. three sera thus obtained were found to be monospecific for fmdv proteins 3a, 3c, and 3d, respectively. two others were prevalently directed against protein 2c, but in addition, either to protein 2b or to protein 3a. five out of six mature nonstructural vi ...19892554586
analysis of foot-and-mouth disease virus-neutralizing idiotypes from immune bovine and swine with anti-murine idiotype antibody probes.rabbit anti-idiotypic antibodies (a-idab) induced by foot-and-mouth disease virus (fmdv) neutralizing mab were used as probes to identify anti-fmdv id in immune serum from bovine and swine. in a competitive ria, at least two of the a-idab exhibited a dose-dependent capacity to compete with labeled virus for anti-fmdv antibodies from a convalescent bovine serum. these a-idab were immobilized on activated sepharose and used to isolate anti-viral id from bovine, swine, and murine fmdv immune sera. ...19892553817
comparison of a liquid-phase blocking sandwich elisa and a serum neutralization test to evaluate immunity in potency tests of foot-and-mouth disease vaccines.sera from cattle vaccinated against either foot-and-mouth disease virus (fmdv) strains a10 holland, o1 bfs, or c1 detmold were tested in a serum neutralization test (snt) and a liquid-phase blocking sandwich elisa (lbe), and the titers were compared with the results of intradermolingual challenge tests. the lbe test results were significantly more reproducible (p less than 0.005) than the snt results. the correlation coefficients between snt and lbe were 0.91 for fmdv strains a10 holland and o1 ...19892553819
a theoretical study of the acidification of the rhinovirus capsid.electrostatic calculations for human rhinovirus 14 indicate that histidine-base residue pairs in the region of a beta-strand interaction between pentamers may be involved in a ph-induced process that leads to the release of viral rna. other picornavirus sequences are examined for these residue pairs, a subset of which is present in enteroviruses. foot and mouth disease virus possesses one of the residue pairs, and cardioviruses, which undergo a separate ph and halide ion-induced capsid dissociat ...19892555222
[antigenic structure of foot-and-mouth virus. iv. synthesis and immunogenic properties of new fragments of the vp1 protein of food-and-mouth virus strain a22].a synthesis of new fragments of vp1 protein with the specificity of a22 strain of foot-and-mouth disease virus is described. immunization with the free 136-152 peptide and klh-conjugates of the peptides 136-152 and 197-213 induced 60-80% protection of guinea pigs against challenge with the a22 virus. synthetic peptides corresponding to the 10-24, 50-69 and 175-189 sequences of vp1 did not show any protective activity. we have found that uncoupled peptides 175-189 and 197-213 are able to induce a ...19892556150
assay sensitivity and differentiation of monoclonal antibody specificity in elisa with different coating buffers.buffers of different ph and ionic strength were employed as coating buffers for antigen adsorption to microtitre plates. their efficiency for coating plates with rinderpest virus (rpv) and foot-and-mouth disease virus (fmdv) antigens was studied by elisa with polyclonal and monoclonal antibody preparations. while the adsorption and detection of rpv antigen with polyclonal antiserum was highly dependent on the ionic strength and ph of coating buffer, adsorption of antigenically active fmdv antige ...19892551906
protection induced by synthetic peptides corresponding to three serotypes in foot and mouth disease virus. 19892558525
hydrolysis of a series of synthetic peptide substrates by the human rhinovirus 14 3c proteinase, cloned and expressed in escherichia coli.the 3c proteins of several picornaviruses, including poliovirus, foot-and-mouth disease virus (fmdv) and encephalomyocarditis virus (emcv), have been demonstrated to be cysteine-type proteinases, involved in the processing of the respective polyproteins expressed by the monocistronic rna genome. nucleotide sequencing data have indicated that the human rhinovirus 14 (hrv-14) rna genome encodes a homologous 3c protein. the hrv-14 3c protein was purified to homogeneity from escherichia coli express ...19892555433
[an immunoenzyme method of isolation of foot-and-mouth disease virus by using beta-lactamase conjugate with virus-specific antibodies].it was shown that in was feasible to use conjugates of virus-specific antibodies and beta-lactamase from bacillus licheniformis 749/c to identify aphthosa virus antigens. the antigen titers determined by enzyme immunoassay (eia) using a beta-lactamase conjugate were 5-64 times higher than the analogous indices of the complement fixation test. unlike eia, that by using the antibody conjugates with peroxidase or alkaline phosphatase there were observed no "background" responses.19892559667
[use of sarcoma 180 tg cells for obtaining ascitic fluid from mice hyperimmune to the foot-and-mouth disease virus].mice immunized with fmdv c3 arg 84 antigen were inoculated intraperitoneally with sarcoma 180 tg. the ascitic fluid obtained by ventral puncture contained high titers of antibodies, similar to those obtained from serum, as determined by neutralization and elisa tests. ascitic volumes were 10 to 20 times greater than those obtainable with.19892559425
antigenic variation of foot-and-mouth disease virus of serotype c during propagation in the field is mainly restricted to only one structural protein (vp1).the primary structure of vp3, vp2 and vp4 capsid protein genes has been determined for six epizootiologically-related foot-and-mouth disease virus (fmdv) isolates of serotype c1, two of which presented immunogenic differences as determined by a cross-protection assay. the results obtained have been compared with those previously reported for the corresponding vp1 genes martinez et al. (1988) gene 62, 75-84. high rates of fixation of mutations have been estimated for the four capsid protein genes ...19892560293
use of in situ hybridization for the detection of foot-and-mouth disease virus in cell culture.biotinylated complementary dna (cdna) and rna probes were prepared from a specific and highly conserved section of the foot-and-mouth disease virus (fmdv) genome coding for the rna-dependent rna polymerase. hybridization was conducted on fmdv-infected, bovine enterovirus (bev)-infected, and noninfected swine kidney cell cultures. the detection system utilized the enzyme system streptavidin-alkaline phosphatase, the substrate phosphate, and the chromogen nitroblue tetrazolium. intense cytoplasmic ...19892562224
survival of foot-and-mouth disease virus in sausage meat products (italian salami).determination of the survival of foot- and-mouth disease virus (fmdv) in fresh meat from experimentally infected swine and in several types of sausage meat (italian salami) produced according to the technology widely applied by the principal italian producers has been carried out. the purpose of the experiment was to assess if typical italian salami can be considered safe with regard to the spread of fmd through international trade. the results obtained showed: (a) high titers of fmdv were detec ...19892561953
[antigenic structure of the foot-and-mouth virus. v. protection of naturally susceptible animals from foot-and-mouth disease using a synthetic peptide].we have synthesized the peptide representing 135-159 vp1 sequence of a22 strain of the foot-and-mouth disease virus (fmdv). the synthetic peptide induced 100% protection of guinea pigs against the disease. two-fold immunization of cuttle with the peptide and single immunization of sheep induced full protection of the animals against a22 strain of fmdv.19892561049
the isoelectrofocusing technique in comparison of some sudanese type sat-1 foot-and-mouth disease viruses.isoelectric focusing technique (ieft) was employed to compare type sat-1 fmd virus from sudan. results of the ieft tests were compared with available previous serological and epidemiological data on the viruses used. possible potential uses of the test, in parallel with previously available serological and epidemiological data, are discussed.19892562038
[immune response against foot-and-mouth disease virus in cattle: effect of vaccination].foot and mouth disease virus (fmdv) is one of the most feared animal virus and vaccination still has to be used in many countries. in previous reports, using a murine model, we studied the cellular basis of immune responses against fmdv and were able to show that they are atypical. in cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against fmdv, except for antibody responses, but practically nothing concernin ...19892562135
[primary structure of the gene for vp1 protein of the foot-and-mouth disease virus of asia 1 serotype].the nucleotide sequence of the cdna for the viral rna region coding for the main antigenic protein of the epidemic stomatitis virus of asia 1 serotype has been identified. the amino acid sequences in the regions of vp1 protein antigenic determinants of the serotype asia 1 virus and other serotypes viruses have been compared.19892561378
hybridoma cell lines secreting monoclonal antibodies to foot-and-mouth disease virus type asia-1.various immunizing regimens, cell culture requirements and cell fusion conditions were examined for efficient production of hybridomas secreting anti-foot-and-mouth disease virus (fmdv) antibodies. a highly sensitive streptavidin-biotin-based enzyme-linked immunosorbent assay (elisa) was used for screening of hybridomas for specific antibody production as well as for determining the serotype specificity of the antibodies. six hybridoma cell lines generating antibodies to fmdv type asia-1 (vaccin ...19892569807
fimbriae of bacteroides nodosus: protein engineering of the structural subunit for the production of an exogenous peptide.the pattern of sequence variation between bacteroides nodosus fimbrial subunits of different serotypes suggests a degree of flexibility, which might be exploited for protein engineering approaches for the expression of other peptides. we have tested this using the well-characterized peptide epitope from vp1 of foot-and-mouth disease virus (fmdv), residues 144-159: lrgdlqvlaqkvartl (strain 01-bfs). using bacterial codon usage, several oligonucleotides were designed for the substitution of this se ...19892564674
translational fusions with fragments of the trpe gene improve the expression of a poorly expressed heterologous gene in escherichia coli.a series of plasmids expressing fusions between the trpe gene product, anthranilate synthase component i and the major immunogen (vp1) of foot and mouth disease virus were constructed such that increasing amounts of the 3' end of trpe were deleted. deletions removing up to 70% of trpe had little effect on the quantity of fusion protein expressed, while the number of molecules appeared to increase. larger deletions led to a steady decrease in both the quantity of fusion protein produced and in th ...19892674321
type 1 fimbriae of escherichia coli as carriers of heterologous antigenic sequences.a strategy has been designed for the construction of recombinant bacterial strains which eventually may become useful as live vaccines and which may also be relevant for the preparation of conventional vaccines. the approach used is the fusion of small antigenic peptide sequences into specific segments of a protein whose location on the bacterial surface ensures that the recombinant organism is able to present the inserted antigen to the host (animal or human) infected by the bacterium. the chos ...19892576014
small peptides induce antibodies with a sequence and structural requirement for binding antigen comparable to antibodies raised against the native protein.antisera were raised against the chemically synthesized peptide corresponding to each epitope of three foot-and-mouth disease virus strains. peptide synthesis was further used to determine which amino acid residues in each epitope are important for the specificity of antisera raised against the whole virus. the specificity of the antibody paratope for its epitope was shown to depend on structure as well as sequence. anti-virus sera demonstrated a greater specificity for the homologous peptide th ...19852578661
generation of a sheep x mouse heterohybridoma cell line (1c6.3a6t.1d7) and evaluation of its use in the production of ovine monoclonal antibodies.a stable aminopterin-sensitive sheep x mouse heterohybridoma cell line (1c6.3a6t.1d7) for use in the generation of sheep monoclonal antibodies is described. the line was first constructed by fusing the mouse myeloma line, nso, to normal sheep lymphocytes obtained from the efferent lymphatic vessel of a cannulated popliteal lymph node. the line was rendered sensitive to aminopterin through a combination of irradiation and treatment with the anti-metabolite drug 6-thioguanine. characterisation of ...19892760466
potential secondary and tertiary structure in the genomic rna of foot and mouth disease virus.the nucleotide sequence of the 5' untranslated region of foot and mouth disease virus (fmdv), serotype a10 has been determined. this completes the first total genomic sequence for any one serotype of fmdv. analysis of the sequence to the 3' side of the poly (c) tract reveals the presence of a 24 nucleotide repeated motif which has homologies with a sequence located upstream of the transcriptional initiation site from several mammalian fibrinogen genes. the function of this element in fmdv is unc ...19872821491
[immunologic relations between the foot-and-mouth disease virus isolates a5 westerwald 1951 and a5 bernbeuren 1984]. 19872820373
detection and typing of foot-and-mouth disease virus by enzyme-linked immunosorbent assay: a sensitive, rapid and reliable technique for primary diagnosis.a highly sensitive indirect sandwich enzyme-linked immunosorbent assay suitable for adoption as the routine diagnostic and typing test for foot-and-mouth disease virus of all seven serotypes is described. the assay uses rabbit and guinea pig antisera raised against inactivated 146s virus antigens. strong homotypic and minimal heterotypic reactions with both whole virion 146s and derived virion subunit 12s antigens achieved a detection sensitivity approximately 125 times that of the complement fi ...19872825310
[assays of cytotoxicity and antiviral activity of crude and semipurified extracts of green leaves of melia azedarach l].crude extracts from fresh green leaves of melia azedarach l contain an antiviral factor (fav) able to inhibit the replication of several animal viruses, e.g. polio, vsv, hsv, fmdv, sindbis, junín, pichinde and tacaribe in vero or bhk-21 cells. crude preparations were subjected to different steps of purification like chromatography on sephadex g-100 and deae-sephadex. the antiviral activity of g-100 and deae fractions was fully conserved, whereas contaminating proteins were lost. two types of cyt ...19852825236
safety and efficacy of foot-and-mouth disease vaccines containing endonuclease-inactivated virions.inactivation of foot-and-mouth disease virus (fmdv) by means of virion-associated endonuclease was found to be suited to the production of safe and potent vaccines, which proved to be equal or better than those containing formaldehyde or ethyleneimine in guinea-pig potency tests. first order inactivation kinetics were regularly shown, with half life values which varied according to the different temperatures used. inactivation brought about extensive degradation of fmdv rna, while it did not adv ...19872823495
experimental infection of eland (taurotrages oryx), sable antelope (ozanna grandicomis) and buffalo (syncerus caffer) with foot-and-mouth disease virus.the course of experimental infection of a type sat 1 fmdv strain was studied in buffalo, sable antelope and eland following tongue inoculation and contact and has been compared with that in cattle. all species became infected, although disease was less severe in the game animals and larger amounts of virus were required to infect game animals than cattle. neutralizing antibody titres were high and were maintained for an extended period in buffalo, sable antelope and eland. the carrier state was ...19892584449
crystallization and preliminary x-ray diffraction analysis of foot-and-mouth disease virus.foot-and-mouth disease virus has been crystallized with the objectives of (1) determining the composition and conformation of the major immunogenic site(s) and (2) comparing its structure with those of the related polio, rhino and mengo viruses, representing the other three genera of the picornaviruses. most of the work has been done with virus strain o1bfs 1860, which crystallized as small rhombic dodecahedra of maximum dimension 0.3 mm. virus recovered from crystals was infectious, and was ind ...19872824786
neutralization sites of type o1 foot-and-mouth disease virus defined by monoclonal antibodies and neutralization-escape virus variants.monoclonal antibodies (mabs) were derived from mice infected with foot-and-mouth disease virus type o1 brugge (fmdv 01b) or immunized with inactivated virions (140 s) or viral subunits (12 s). a total of 19 neutralizing mabs were characterized of which 17 recognized conformationally determined epitopes and two recognized amino acid sequences on isolated vp1. neutralizing mabs were used to select antigenic variants of fmdv o1b. based on cross-neutralization and binding assays with mabs the varian ...19882827379
immunoglobulin profiles in nasal and buccal secretions from normal crossbred calves after vaccination with inactivated virus and/or experimental exposure to foot-and-mouth disease virus type asia i. 19872827446
[in vitro translation of foot-and-mouth disease virus ribonucleic acid]. 19872829771
response to foot-and-mouth disease vaccines in newborn calves. influence of age, colostral antibodies and adjuvants.oil-emulsified (oe) and aqueous (aq) vaccines were prepared with the same batch of inactivated a24 8345 foot and mouth disease virus (fmdv). calves born to vaccinated dams did not respond to the aq vaccine 30 or 90 days post partum. when the oe vaccine was used on a similar group of calves, no responses were elicited up to 21 days post partum. however, calves 30 or more days old responded like adult cattle to the oe vaccine. when the oe vaccine was used in colostral antibody-free calves 3-30 day ...19882828089
[preparation of the 35s ribonucleic acid of foot-and-mouth disease virus]. 19872829770
[differentiation of foot-and-mouth disease viruses by counterimmunoelectrophoresis].studied were the opportunities for employing the method of counter immunoelectrophoresis (cie) to differentiate f. m. d. viruses of different origin, comparing the results obtained with those reached through the immunodiffusion test (idt). it was found that cie could be used successfully for the serotyping of f. m. d. viruses, the method being as precise as idt, however, the results are recorded at the 90th minute, while idt requires at least 24 hours to read the results.19872830709
[cdna cloning of foot-and-mouth disease virus]. 19872829769
vp1 of serotype c foot-and-mouth disease viruses: long-term conservation of sequences.the nucleotide sequences of the vp1-coding regions of several isolates of serotype c3 foot-and-mouth disease virus (fmdv) were determined. the deduced amino acid sequences were compared with those of serotype c1 fmdv. the results provide evidence for two different lineages of fmdv c3 and document the potential for both long-term conservation and rapid evolution of fmdv.19882831408
[quantitative studies of ammonium sulfate precipitation of foot-and-mouth disease virus]. 19872831829
[diagnosis of foot-and-mouth disease and strain identification of the foot-and-mouth disease virus]. 19872831824
[foot-and-mouth disease virus replication in cloned bhk-21 cell cultures with reduced serum requirements]. 19872831826
cell mediated immunity to foot and mouth disease virus vaccine in buffaloes. 19872831140
rapid selection of genetic and antigenic variants of foot-and-mouth disease virus during persistence in cattle.rapid evolution of foot-and-mouth disease virus (fmdv) is documented during persistent infections of cattle. the carrier state was established experimentally with plaque-purified fmdv of serotype c3. virus was recovered from the esophageal pharyngeal area of the animals up to 539 days postinfection. analysis of capsid proteins by electrofocusing and by electrophoretic mobility of the genomic poly(c)-rich tract suggested heterogeneity in several isolates and sequential dominance of viral subpopul ...19882835508
coevolution of cells and viruses in a persistent infection of foot-and-mouth disease virus in cell culture.virus and cells evolve during serial passage of cloned bhk-21 cells persistently infected with foot-and-mouth disease virus (fmdv). these carrier cells, termed c1-bhk-rc1 (j.c. de la torre, m. dávila, f. sobrino, j. ortín, and e. domingo, virology 145:24-35, 1985), become constitutively resistant to the parental fmdv c-s8c1. curing of late-passage c1-bhk-rc1 cells of fmdv by ribavirin treatment (j.c. de la torre, b. alarcón, e. martínez-salas, l. carrasco, and e. domingo, j. virol. 61:233-235, 1 ...19882835509
infection of cattle by airborne foot-and-mouth disease virus: minimal doses with o1 and sat 2 strains.equipment has been constructed and methods developed for exposing individual cattle to two strains of foot-and-mouth disease (fmd) virus in aerosols to determine the minimal infective dose by the respiratory route. the aerosols used were produced either artificially by a spinning-top aerosol generator, in which case they were of homogeneous small particle size (less than 3 micron in diameter) or else they were derived naturally from infected pigs, in which case the particles were heterogeneous i ...19872832913
detection of foot-and-mouth disease virus with dna probes in bovine esophageal-pharyngeal fluids.infectivity and dot-blot hybridization techniques were compared for the detection of fmdv in esophageal-pharyngeal fluids from experimentally infected cows. the probe used includes the viral polymerase sequence which allows the detection of the three types of virus (a, o, and c) with equivalent sensitivity. virus was detected by dot-blot hybridization as well as by infectivity, according to sample analysis of esophageal-pharyngeal fluids extracted seven days post-infection. it was not possible t ...19882833204
new approaches to animal vaccines utilizing genetic engineering.control of infectious diseases in livestock is an important determinant in the success of a nation's effort to efficiently meet its need for animal products. genetic engineering offers many new options in the design of animal vaccines. monoclonal antibodies, dna cloning, recombination, and transfection are examples of techniques that facilitate innovative strategies in antigen identification, production, and delivery. this article reviews the use of genetic engineering in the production of vacci ...19882837363
analysis of neutralizing epitopes on foot-and-mouth disease virus.for the investigation of the antigenic determinant structure of foot-and-mouth disease virus (fmdv), neutralizing monoclonal antibodies (mabs) against complete virus were characterized by western blot (immunoblot), enzyme immunoassay, and competition experiments with a synthetic peptide, isolated coat protein vp1, and viral particles as antigens. two of the four mabs reacted with each of these antigens, while the other two mabs recognized only complete viral particles and reacted only very poorl ...19882835507
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