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PubMed1986

Synthesis of fusion proteins containing antigenic determinants of foot-and-mouth disease virus.

Part of the genome of foot-and-mouth disease virus (FMDV) type 01,BFS, including the sequence encoding the capsid polypeptide VP1, was cloned in Escherichia coli following a new cloning strategy. The clone containing the VP1 sequence was used for the construction of two expression plasmids encoding VP1 fusion proteins. Subsequently, substantial amounts of the two VP1-beta-galactosidase fusion proteins, containing either one (amino acid region 140-160) or two (amino acid regions 140-160 and 200-213) antigenic determinants of the virus, were synthesized by E. coli bacteria. The protein containing the amino acid region 140-160 of VP1 fused to beta-galactosidase efficiently induced antibodies in rabbits specifically reacting with FMDV type 01,BFS. The same protein was also capable of eliciting neutralizing antibodies. The fusion protein containing both antigenic determinants did not efficiently induce antibodies reacting with FMDV.

Animals↗
PubMed1997

Characteristic in vitro evolution pattern of foot and mouth disease virus A81/Castellanos/Arg/87.

The in vitro evolution of Foot and mouth disease virus (FMDV) A/81/Castellanos/Arg/87 (A/Castellanos/87) was studied by partial biological and biochemical characterization of viral populations selected after 25 passages on secondary fetal bovine kidney cell monolayers. These passages were performed in the presence or absence of immune pressure exerted in the form of antiviral polyclonal serum. While the viral populations passaged in the absence of immune pressure acquired characteristics such as antigenic heterogeneity, VP1 amino acid modification and plaque size reduction, the populations selected after immune pressure also presented both neutralizing resistance and attenuation for suckling mice. The comparison with other previously studied FMDV strains suggests that FMDV A/Castellanos/87 adopts a differential response to immunological pressure and other selective forces. In addition, the sequencing analysis of viral selected populations shows a restriction in the number and type of amino acid replacements tolerated by FMDV capsid proteins.

Amino Acid Sequence↗
PubMed1978

Early events in the interaction between foot-and mouth disease virus and primary pig kidney cells.

Foot-and-mouth disease virus (FMDV) attached to pig kidney cells at 0 degrees C and could only be recovered in a form with a sedimentation coefficient and buoyant density lower than that of the native virus. Incubation of the virus-cell complex at 37 degrees C caused disruption of about 80% of the particles into a 12S protein sub-unit that had the same polypeptide composition as that produced by reducing the pH of the virus below pH7. The remaining 20% had the same polypeptide and RNA composition as the native virus but it had a lower sedimentation coefficient, buoyant density and specific infectivity. These lower values are probably due to the association of the virus with cell membrane components. The 12S subunits were shown to be located inside the cell, indicating that disruption of the virus had occurred within the cell. The results are discussed in relation to the different cell mediated alteration of other picornaviruses.

Adsorption↗
PubMed1985

Analysis of the secondary structure of the poly(C) tract in foot-and-mouth disease virus RNAs.

Sodium bisulphite modification of foot-and-mouth disease virus (FMDV) RNA in solution indicates that the majority of the poly(C) tract in the RNA is single-stranded in concordance with previous results with encephalomyocarditis virus RNA. The reaction kinetics are biphasic; 60% of the cytidylic acid in the poly(C) tract reacts like synthetic poly(C), and the remainder with the kinetics of the cytidylic acid in the rest of the RNA. The reactivity of the poly(C) tract with poly(I) indicates that it is looped out and exposed in the RNA. The deamination reaction has also been used to investigate the structure of the replicative form (RF) and replicative intermediate (RI) isolated from infected cells. Analysis by gel electrophoresis of the long RNase A- and T1-resistant oligonucleotides of RI suggests that it has five single-stranded poly(C) tracts to every one which is base-paired. Bisulphite reactivity of the poly(C) tract and gel electrophoresis of the ribonuclease-resistant oligonucleotides of RF indicate that the poly(C) is base-paired to a poly(G) tract in this molecule. The presence of a poly(G) tract in RF and RI provides unequivocal evidence that the poly(C) is replicated via poly(G) in the negative strand.

Animals↗
PubMed1990

Heterotypic recognition of foot-and-mouth disease virus by cattle lymphocytes.

Lymphoproliferation against foot-and-mouth disease (FMD) virus was examined using peripheral blood mononuclear cells from vaccinated cattle. Ten weeks after revaccination the optimum conditions for proliferation were obtained with 1 microgram/ml of purified virus after 5 to 6 days in culture. This contrasted with the response at 20 months post-revaccination, when the response required less antigen and showed a peak response after 3 to 4 days in culture. Proliferation was specific for FMD virus, but was cross-reactive between serotypically distinct strains of the virus. The proliferative response to isolated virus proteins (VP) involved all three major capsid proteins (VP1, -2 and -3), although the proliferation of lymphocytes from heterotypically vaccinated cattle was due to VP3. Furthermore, the response induced by purified virus, chemically fixed virus and subunit virus particles was indistinguishable and thus it is likely that processing was required for the induction of proliferation. Together these data strongly suggest that FMD virus-induced lymphoproliferation is T cell-mediated and that VP3 may contain dominant, cross-reactive sequences.

Animals↗
PubMed1967

Chemically characterized media for study of foot-and-mouth disease virus in baby hamster kidney cells.

Foot-and-mouth disease virus can be grown in baby hamster kidney cells with a chemically characterized medium containing only tris(hydroxymethyl)-amino-methane (Tris) buffer, glucose, glutamine, and salts. Virus infectivity was only 0.5 log unit less than in a complex cell growth medium containing serum, tryptose phosphate, and lactalbumin hydrolysate. At high multiplicity of infection, production was maximal in 5 hr, with the virus remaining largely intracellular. Glucose and glutamine appeared to act independently of each other although both were required at about the same time during the virus production cycle. Glutamine had the greater effect and could not be replaced by amino acids, purines, and pyrimidines. Glutamine also stimulated cellular oxygen uptake in both normal and infected cells. Serum and other organic components added singly to the defined medium did not increase the virus yield. Studies on uninfected cells over a 5-hr incubation period showed that the defined medium maintained protein and ribonucleic acid synthesis at rates similar to the complex cell growth medium. These rates were much lower in media containing only inorganic salts and Tris buffer. Glucose, however, was more important to uninfected cellular metabolism than was glutamine. Defined medium containing dialyzed calf serum produced the highest rate of protein synthesis.

Amino Acids↗
PubMed1976

[Immunogenic quality of foot-and-mouth disease virus].

Routine analysis of suspensions of foot-and-mouth disease virus and eluates of vaccine by the isopycnic method in analytical ultracentrifuge demonstrates the important heterogeneity of the viral population. This heterogeneity increases during inactivation of the virus by formol. In view of this dispersion of the physical characteristics of viral particles it may be asked (a) whether the immunogenic value is linked to the total quantity of particles which, we know, are very different from each other and of which we can determine only the partial or total parameters or (b) whether, on the contrary, the immunogenic value is linked to the activity of marginal particles, very small in number, and perhaps even quantitatively indistinguishable by the physical methods available. However, the hypothesis of particles possessing different properties but converging in the final result cannot, a priori, be excluded. Consequently it cannot be hoped that a simple relation between the concentration in weight of a viral suspension and its immunogenic property may be determined.

Antigens, Viral↗
PubMed2004

A first molecular epidemiological study of SAT-2 type foot-and-mouth disease viruses in West Africa.

Thirty-one viruses causing SAT-2 outbreaks in seven West African countries between 1974 and 1991, and four viruses representative of East and Central Africa were genetically characterized in this study. Four major viral lineages (I-IV) were identified by phylogenetic analysis of an homologous 480 nucleotide region corresponding to the C-terminus end of VP1. Lineage I comprised two West African genotypes with viruses clustering according to year of isolation rather than geographical origin. Lineage II was represented by viruses isolated between 1979 and 1983 in two neighbouring West African countries, Senegal and The Gambia. Viruses from Nigeria and Eritrea, representative of West and East Africa respectively, constituted lineage III, whilst lineage IV, comprising viruses from Central and East Africa, was regionally and genetically distinct. This study revealed that unrestricted animal movement in West Africa is a major factor in disease dissemination and has also provided the first indication of trans-regional virus transmission.

Africa, Western↗
PubMed2004

Differentiating infection from vaccination in foot-and-mouth-disease: evaluation of an ELISA based on recombinant 3ABC.

Recent devastating outbreaks of foot-and-mouth disease (FMD) in Europe have reopened the discussion about the adequacy of the non-vaccination strategy implemented by the EU in 1991. Here we describe the evaluation of a new commercially available test kit for the discrimination between vaccination and infection. The test is based on the detection of antibodies against the recombinant non-structural (NS) protein 3ABC. In contrast to immunization with vaccines free of 3ABC, these antibodies are elicited as a consequence of infection. Testing more than 3600 negative sera from several countries revealed a specificity of > 99% for bovine, ovine, and porcine samples. Antibodies specific for 3ABC can be detected as soon as 10 days post-infection. As compared with the occurrence of antibodies against structural proteins of FMDV, anti-3ABC antibodies can be detected 5-10 days later, depending on the species. No anti-3ABC antibodies were detected in sera from vaccination experiments or in field sera from vaccinated animals. However, anti-3ABC antibodies can be detected in vaccinated animals upon challenge. These results provide evidence that this test can facilitate the use of vaccines in new strategies against FMD.

Animals↗
PubMed2000

Carriers of foot-and-mouth disease virus: a review.

This review describes current knowledge about persistent foot-and-mouth disease virus (FMDV) infections, the available methods to detect carrier animals, the properties of persisting virus, the immunological mechanisms, and the risk of transmission. In particular, knowledge about the carrier state, the period in which virus can be isolated from animals 28 days or longer post infection, is important, because the risk that animals may carry the virus will influence the diagnostic and preventive measures that need to be taken. Although many years of research have led to much knowledge about foot-and mouth disease and its causative agent, there are still numerous aspects of the virus and the disease that are not yet fully understood. Areas for further research on persistence of FMDV are discussed.

Animals↗
PubMed1987

Liposome encapsulated subunit (VP1) and virion vaccines against foot-and-mouth disease.

Subunit vaccine prepared from VP1 protein of foot-and-mouth disease virus (FMDV) types 0 and Asia 1 protected guinea pigs against FMD and also induced high levels of antibody. Liposomes have been used as a safe and potent immunological adjuvant for FMD vaccines. Vaccines prepared from inactivated virus types 0 and Asia 1 encapsulated in liposomes protected guinea pigs against challenge with homologous virus and showed good antibody response in pigs on a small scale field trial.

Adjuvants, Immunologic↗
PubMed1990

Protection of cattle and swine against foot-and-mouth disease, using biosynthetic peptide vaccines.

A single dose of foot-and-mouth disease (FMD) virus protein 1 (VP1) peptide, expressed in Escherichia coli as a fusion protein with 190 amino acids (AA) of the LE' protein of the tryptophan operon of E coli, elicited an immune response in steers sufficient to withstand the challenge of exposure to animals with acute FMD. The 58-micrograms dose of viral peptide, composed of a segment of the VP1 from the A12 strain (A12) of FMD virus (FMDV; A12-32dimer) in a tandem repeat configuration of AA137 through 168 and emulsified with oil adjuvant, elicited a serologic response in cattle equivalent to that obtained using conventional whole virus vaccines. Two groups of swine were vaccinated, 1 with the A12-32dimer as used in cattle and 1 with AA131 through 157 from VP1 of the A24 strain (A24) of FMDV (A24-peptide), expressed in the same system as A12-32dimer, but as a single copy per molecule. In swine, the 58-micrograms dose of the A12-32dimer repeated at 28 days was an effective immunogen; all swine were protected against A12 and, in addition, the vaccine protected 50% of the swine against A24. The 29-micrograms dose of A24-peptide, administered according to the same schedule, elicited protection against A24 in 50% of the vaccinates and, in addition, protected 25% of those vaccinates against A12. The serologic response elicited by A12-32dimer against A24 virus was considerably greater than the response elicited by A24-peptide against A12 virus. The evidence of multiple immunogenic epitopes between AA131 and AA168 was evaluated.

Amino Acid Sequence↗