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PubMed1998

Analysis of the B and T cell response in guinea pigs induced with recombinant vaccinia expressing foot-and-mouth disease virus structural proteins.

Recombinant vaccinia viruses expressing foot-and-mouth disease virus (FMDV) P1 and VP1 genes have been used to study the immune response induced by these viral polypeptides in guinea pigs. Anti-FMDV antibodies, but not neutralizing activity, were detected in the sera from immunized animals. The results indicate that both CD4+ and CD8+ FMDV-specific T cells were induced by the vaccinia recombinants. Consistently with the activation of CD4+ T cells, lymphocytes from immunized animals specifically proliferated in vitro in response to whole virus. The induction of virus-specific CD8+ T cells was determined by CTL assay of immune splenocytes restimulated in vitro with FMDV infected cells. Altogether, the results obtained indicate that both B and T cell immune responses to FMDV are elicited upon immunization of guinea pigs with vaccinia recombinants expressing FMDV structural polypeptides.

Animals↗
PubMed1990

Mouse footpad Langerhans cells as an indicator for safety of foot and mouth disease virus vaccines.

The effect of various vaccines against foot and mouth disease virus (FMDV) was tested on Langerhans cell density in the footpad epidermis of mice. Injection of monovalent, bivalent and trivalent FMDV vaccines caused a reduction in Langerhans cell density in the murine skin, which was more marked at the center of the footpad, the site of injection, than at the periphery. Testing of the various components of the vaccine showed that saponin caused a marked reduction in Langerhans cells while injection of aluminium hydroxide had a lesser effect and the virus alone had no effect on these cells. Thus Langerhans cell density could serve as an efficient marker to test the safety of vaccines to FMDV since the integrity of Langerhans cells, which are the antigen-presenting cells in the skin epidermis, is needed for an effective immune response to the vaccine.

Adenosine Triphosphatases↗
PubMed1989

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 VP1. Another epitope originally thought to occupy a single site in the area of amino acids 168-179 of VP1 was found on all of the variants. Competitive binding assays did not identify the exact binding site for this monoclonal antibody and the results suggest that the epitope may represent a site more complex than a sequential epitope. A polyclonal antiserum to a 13 kDa fragment of VP1 (residues 55-179) was found to have all the virus reactivity associated with sites located between residues 133-164 of VP1. In contrast, an antiserum to VP1 was found to have additional virus binding sites outside of the 133-164 region of VP1.

Antibodies, Monoclonal↗
PubMed1980

Nucleotide sequence heterogeneity of the RNA from a natural population of foot-and-mouth-disease virus.

The genomic RNA from isolates of foot-and-mouth-disease virus (FMDV) of serological types O or C obtained during epizootic outbreaks have been analysed by two-dimensional gel electrophoresis of the T1 RNase-generated oligonucleotides (T1 fingerprinting). Among virus isolates that are closely related serologically, 4-12 oligonucleotide changes were detected constitute the genome, the variations affect 0.7%-2.2% positions in FMDV RNA. Higher nucleotide-sequence divergence exists between the genomic RNAs from serologically unrelated viruses, while a 100-fold lower RNA sequence heterogeneity has been detected by analysis of individual clones derived from one viral isolate. Oligonucleotide mapping indicates that the variant oligonucleotides are scattered throughout the FMDV genome. We suggest that extensive genetic variability at many RNA sites is the basis for the antigenic diversity of FMDV.

Antigens, Viral↗
PubMed1984

Purification and immunogenicity of fusion VP1 protein of foot and mouth disease virus.

A procedure has been developed to purify foot and mouth disease virus (FMDV) VP1 surface antigens from recombinant Escherichia coli. The VP1 antigens are expressed as fusion proteins derived from the E. coli Trp operon and VP1 surface protein of FMDV. The procedure is capable of recovering greater than 96% of the desired product at a purity of greater than 96%. The resulting antigens induce significant levels of virus-neutralizing antibody in guinea pigs and cattle as determined by a mouse protection assay [Skinner, H.H. (1952) Proc. Int. Vet. Congr., 15th 1, 195]. E. coli contaminants have a deleterious effect on ion-exchange chromatography as well as immunogenicity of the expressed fusion VP1 antigens. The method presented removes significant E. coli contaminants, yielding fusion VP1 proteins which are immunogenically potent. In particular, virus neutralization titers at 100-micrograms dosage of the fusion VP1 proteins of the O1 and A24 serotypes are similar to that induced by the natural VP1 proteins isolated from FMD virions.

Antigens, Viral↗
PubMed1980

Biochemical aspects of variation in foot-and-mouth disease virus.

The biochemical basis for variation in foot-and-mouth disease virus (FMDV) has been explored by analysis of the virus RNA and the virus-induced and structural proteins of three isolates of the virus. Two of the isolates were from serotype A and the third was from serotype O. Hybridization studies of the RNAs showed greater than 80% homology between the two type A viruses and about 65% homology between the two type A viruses and the virus of type O. The ribonuclease T1 maps of the three viruses gave distinct patterns typical of FMDV, but did not show that any two of the three viruses were more closely related. The virus-induced primary translation products, P88, P52 and P100 isolated from infected cells, were compared by tryptic peptide analysis. Combinations of 3H- and 14C-leucine-labelled polypeptides were hydrolysed with trypsin and resolved on an ion-exchange column. Much greater differences were found in P88 than in P52 or P100, indicating that the major variation occurs in the region of the genome coding for the structural proteins. Similar analysis of combinations of the structural proteins of the three viruses showed that there were differences in VP1, VP2 and VP3 and these results were supported by those obtained by PAGE analysis of the Staphylococcus aureus V8 protease cleavage products.

Aphthovirus↗
PubMed1991

Comparison of the immune response elicited by infectious and inactivated foot-and-mouth disease virus in mice.

The immune response to foot-and-mouth disease virus (FMDV) elicited by infection or immunization with inactivated virus in adult mice was examined. A model of adoptive transfer of immunocompetent cells was used for this purpose. The results presented here indicate that both short- and long-term secondary immune responses elicited by high doses of inactivated virus are indistinguishable, at the humoral or cellular level, from that observed after infection. The responses to inactivated or infectious virus were both efficiently mediated by B cells. However, immunization with low doses of inactivated virus induced a response which, although effective in aborting infection, was fully dependent on FMDV-specific T cell cooperation. These findings suggest that the different immune responses observed after infection and immunization are mainly the result of the different viral mass presented to the immune system in each case.

Animals↗
PubMed1977

Evidence for a mouse pathogenicity locus in certain temperature-sensitive mutants of foot-and-mouth disease virus.

Serial tissue culture passaging of three foot-and-mouth disease temperature-sensitive mutants demonstrated the stability of their temperature sensitivity and mouse avirulence characteristics. Recovery of mouse-virulent temperature-sensitive viruses after passage of the mutants in mice suggested that these were not covariant expressions of the same locus, but were under the control of different genes.

Animals↗
PubMed1980

Heterogeneity of the genome-linked protein of foot-and-mouth disease virus.

The genome-linked protein of foot-and-mouth disease virus was examined by electrofocusing in polyacrylamide gels. Two proteins of different charge and amino acid composition were found. The tryptic peptide maps of the proteins were dissimilar. The possible relationship between the two proteins is discussed.

Aphthovirus↗
PubMed1998

Multiple virulence determinants of foot-and-mouth disease virus in cell culture.

Hypervirulent variants of foot-and-mouth disease virus (FMDV) of serotype C arise upon serial cytolytic or persistent infections in cell culture. A specific mutation in the internal ribosome entry site of persistent FMDV was previously associated with enhanced translation initiation activity that could contribute to the hypervirulent phenotype for BHK-21 cells. Here we report that several hypervirulent FMDV variants arising upon serial cytolytic passage show an invariant internal ribosome entry site but have a number of mutations affecting structural and nonstructural viral proteins. The construction of chimeric type O-type C infectious transcripts has allowed the mapping of a major determinant of hypervirulence to the viral capsid. Tissue culture-adapted FMDV displayed enhanced affinity for heparin, but binding to cell surface heparan sulfate moieties was not required for expression of the hypervirulent phenotype in Chinese hamster ovary (CHO) cells. Virulence was identical or even higher for glycosaminoglycan-deficient CHO cells than for wild-type CHO cells. FMDV variants with decreased affinity for heparin were selected from a high-binding parental population and analyzed. Substitutions associated with decreased heparin binding were located at positions 173 of capsid protein VP3 and 144 of capsid protein VP1. These substitutions had a moderate effect on virulence for BHK-21 cells but completely abrogated infection of CHO cells. The comparative results with several FMDV isolates show that (i) increased affinity for heparin and alterations in cell tropism may be mediated by a number of independent sites on the viral capsid and (ii) the same capsid modifications may have different effects on different cell types.

Amino Acid Substitution↗
PubMed2001

Identification of T-cell epitopes in nonstructural proteins of foot-and-mouth disease virus.

Porcine T-cell recognition of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP) was tested using in vitro lymphoproliferative responses. Lymphocytes were obtained from outbred pigs experimentally infected with FMDV. Of the different NSP, polypeptides 3A, 3B, and 3C gave the highest stimulations in the in vitro assays. The use of overlapping synthetic peptides allowed the identification of amino acid regions within these proteins that were efficiently recognized by the lymphocytes. The sequences of some of these antigenic peptides were highly conserved among different FMDV serotypes. They elicited major histocompatibility complex-restricted responses with lymphocytes from pigs infected with either a type C virus or reinfected with a heterologous FMDV. A tandem peptide containing the T-cell peptide 3A[21-35] and the B-cell antigenic site VP1[137-156] also efficiently stimulated lymphocytes from infected animals in vitro. Furthermore, this tandem peptide elicited significant levels of serotype-specific antiviral activity, a result consistent with the induction of anti-FMDV antibodies. Thus, inclusion in the peptide formulation of a T-cell epitope derived from the NSP 3A possessing the capacity to induce T helper activity can allow cooperative induction of anti-FMDV antibodies by B cells.

Amino Acid Sequence↗
PubMed1976

Typing foot-and-mouth disease virus by fluorescent antibody technique.

Typing of foot-and-mouth disease (FMD) virus was performed by the direct fluorescent antibody (FA) technique. Type-specific FA was prepared from the following two sorts of procedures: (1) FA against live virus (FA-live) was prepared from hyperimmune serum taken from guinea pigs having received live FMD virus. Then it was adsorbed with concentrated heterotype antigen. (2) FA against inactivated virus (FA-Inact) was prepared from antiserum taken from guinea pigs immunized with purified FMD virus inactivated with acetylethyleneimine. Seventeen strains of FMD virus (seven strains of type A, seven strains of type O, and three strains of thpe C) were used. Type-specific FMD virus antigen was detected distinctly from the monolayer of BHK cells infected with each type of virus and fixed in acetone, in spite of negative results obtained from the cells fixed in methyl alcohol. All the 17 strains were typed successfully by the implementation of these two FA methods.

Aphthovirus↗
PubMed1976

Serological and immunological relationships among type A foot-and-mouth disease strains in South America.

Field strains of foot-and-mouth disease (FMD) virus which differ immunogenically from vaccine strains appear from time to time and may present serious problems for FMD campaigns. In order that appropriate measures can be taken it is important to determine the degree of differences between the vaccine strain and the variant strain soon after its appearance. With the A strains studied it was observed that a serological relationship by complement fixation test of greater than or equal to 0.40 would indicate sufficient protection of the vaccine strain for the field strain. With a lower relationship, however, the vaccine strains usually do not fully protect against the field strain. The mouse protection test can be used to determine how serious the lack of protection may be. With this test routine it is possible to issue alerts for intensified epidemiological surveillance within 2-3 weeks after the field strain is received at the Center.

Animals↗
PubMed1967

Effect of incubation temperature and serum content in agar overlay on plaque production by foot-and-mouth disease virus.

The numbers of plaques produced by foot-and-mouth disease virus in primary cultures of calf kidney cells conditioned to 24, 30, and 37 C were essentially the same if virus was absorbed at 37 C. Adsorption was as effective at 30 as at 37 C in cultures conditioned to the respective temperature, but 24 C was less effective under this condition. However, when the adsorption temperature for cultures conditioned to 37 C was decreased to 30 or 24 C, fewer plaques were produced than in cultures conditioned to and maintained at the lower temperatures during absorption. Comparison of the number of plaques produced in cultures overlaid with nutrient agar containing from 0.5 to 10% bovine serum revealed no difference in relation to serum concentration. However, plaque size was related directly to the concentration of serum in the overlay.

Agar↗
PubMedDate not supplied

[Effect of the contrast conditions using solutions of phosphotungstic acid on the electron microscopic image of the foot-and-mouth disease virus].

A method of negative staining of foot-and-mouth disease virus preparations permits to obtain separately positive (2% phosphotungstic acid solution, pH 3.0) and negative (2% PTA solution, pH 6.8 +/- 8.0) stainings. When a 3--4% PTA solution, pH 6.8 +/- 8.0 is used, simultaneous positive and negative staining of each virion is possible which characterizes the functional heterogeneity of the virion protein membrane in interaction with PTA anions.

Aphthovirus↗
PubMed2002

Temporal and geographical distribution of cases of foot-and-mouth disease during the early weeks of the 2001 epidemic in Great Britain.

Estimates of the likely dates of infection of the early cases of the 2001 foot-and-mouth disease (FMD) epidemic indicate that at least 57 premises in 16 counties in Great Britain were infected before the first case was disclosed. Nationwide animal movement controls were imposed within three days of the first case being confirmed on February 20, when FMD was only known to be in two counties, and these controls limited its geographical spread. After the first few cases were confirmed, new cases were rapidly discovered, and the epidemic curve for the daily number of confirmed cases peaked five weeks later, 11 days later than the peak of the curve based on the estimated dates of infection. In the peak week, both curves showed an average daily number of 43 new cases. The estimated dates of infection are believed to be relatively unbiased for the early cases, for which they were derived from a known contact with infection. However, for the later cases they were estimated mainly from the age of the clinical signs of the disease, and were biased by species and other factors, a bias which would probably have made the estimated dates later than was in fact the case.

Animals↗
PubMed1989

Simultaneous vaccination of piglets against foot-and-mouth disease and classical swine fever.

Six-week-old piglets, born of unvaccinated sows, were vaccinated against foot-and-mouth disease (FMD) with a trivalent, inactivated vaccine containing an adjuvant or vaccinated against classical swine fever (CSF) with a live attenuated vaccine or against both diseases simultaneously at two different sites. The antibody response to the FMD vaccine was not significantly influenced by the simultaneous vaccination against CSF. FMD vaccine administered simultaneously with the CSF vaccine produced a significantly higher antibody response to CSF than occurred with CSF vaccination only.

Animals↗