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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↗

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↗

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↗

[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↗

[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.

Amino Acid Sequence↗

Foot and mouth disease virus production on microcarrier-grown cells.

The industrial usefulness of the production of foot and mouth disease virus with microcarrier grown cells has been tested at a large scale. The vaccines, intended for pigs, prepared with such virus give a good level of immunity against virulent challenges. They are stable and free from side effects.

Animals↗

[Study of the immunity in swine vaccinated with type C anti-foot-and-mouth disease vaccine].

Experiments were carried out to adapt a cell culture strain of the foot-and-mouth disease virus, type C, to the organism of susceptible pigs. It was established that 4 to 5 passages are needed to adapt the virus, all treated animals showing the symptoms of the disease from the 24 hour following infection which assumed a generalized course. In determining the index of protection (P) of a given F.M.D. vaccine through challenging immunized pigs its value proves to be 4.5. It is stressed that this is a suitable and economic method to evaluate the immunogenic properties of F.M.D. vaccines for pigs.

Animals↗

Guillain-Barré syndrome following hand-foot-and-mouth disease.

We describe a patient who developed Guillain-Barre syndrome (GBS) following hand-foot-and-mouth disease (HFMD) which is known to be caused by enterovirus infection. A 35-year-old man developed acute paraparesis and dysesthesia in the four limbs following typical skin eruption of HFMD. Electrophysiologic studies showed peripheral nerve demyelination predominant in the distal terminals. HFMD is a rare cause of meningitis, encephalitis, and polio-like myelitis, but GBS following HFMD has never been described.

Adult↗

Lessons from the foot and mouth disease outbreak in The Netherlands in 2001.

The Netherlands had recently developed a new strategy for the eradication of foot and mouth disease (FMD). When FMD was confirmed in Great Britain and France, recent imports of susceptible animals from these countries were traced and preventive measures were taken. On 21 March 2001, FMD was confirmed in The Netherlands. The disease was introduced by calves which became infected at a staging post in Mayenne, France, where infected sheep from Great Britain were present. A total of 26 farms were infected. Emergency vaccination of all susceptible animals was applied. Suppressive vaccination was chosen, implying that all vaccinated animals had to be slaughtered. Ring vaccination of all susceptible animals within 2 km of an infected herd was the standard procedure. However, in the 'Noord Veluwe', vaccination had to be applied to a larger area. The last affected farm was confirmed on 22 April 2001. Emergency vaccination contained the FMD infection rapidly. The last vaccinated animal was slaughtered on 25 May 2001. Many farmers were not convinced that the killing of their healthy, vaccinated animals was justified and tried to prevent the culling, but without success. Politicians and the public at large are now strongly opposed to the large-scale slaughter of vaccinated animals should a future outbreak of FMD occur. The Office International des Epizooties (OIE: World organisation for animal health) should incorporate control of vaccinated animals with non-structural protein (NSP) tests in the chapter on FMD in the International Animal Health Code.

Animals↗

[Mouth diseases and dental materials: potential for study with cytofluorometry].

BACKGROUND: The potential dental materials have of inducing mouth diseases is well known, and various methods have been developed to investigate this phenomenon. Among these there are histological studies of the pulp and periodontium exposed to dental materials, both in vivo and in vitro in humans and animals. Other studies are based on the clinical observation of the effects induced by dental materials. Aim of the paper is to evaluate the use of flow cytometry to analyse crevicular fluid to study its content in terms of inflammatory cells and inflammation mediators. METHODS: Samples of crevicular fluid were collected by aspiration using a bevelled needle mounted on a 5 ml syringe from patients without periodontitis and with periodontitis and multiple heterogeneous dental restorations. This method was adopted since it allows to place the cells of the fluid in suspension. Part of the fluid was analysed by cytology and part by flow cytometry. RESULTS: In the patients without periodontal disease cytological examination revealed the presence of desquamated epithelial cells and colonies of cocci, rods and spirochaetae. In the patients with periodontal disease the same examination revealed, besides the above, the presence of inflammatory cells. Flow cytometry confirmed the findings of cytological examination, thus proving to be an effective method for studying crevicular fluid. CONCLUSIONS: This method allows to identify specific inflammatory cells or mediators of inflammation in crevicular fluid and may therefore prove to be very useful in the study of alterations induced by some dental materials at the level of the gingival sulcus.

Adult↗

Peripheral blood concentrations of plasma steroids and a metabolite of prostaglandin F2 alpha in pregnant cows vaccinated against foot and mouth disease.

Blood samples were collected from 82 pregnant cows at 6-h intervals to determine the effects of vaccination against foot and mouth disease (FMD) on plasma concentrations of progesterone, total unconjugated oestrogens, total corticosteroids and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM). Twenty-two (26.8%) cows showed a strong post-vaccinal reaction in the form of a significant (P < 0.05) elevation in rectal temperature, accompanied by significant (P < 0.01) increases in plasma progesterone and corticosteroid concentrations for 36-42 h after vaccination. Seven (8.5%) cows returned to oestrus within 5-6 days when vaccinated during the first month of pregnancy and nine (10.9%) gave birth prematurely within 72-80 h when vaccinated during the last month of gestation. The importance of post-vaccinal reactions on embryonic mortality and premature birth is emphasized. It is suggested that cows in early and late pregnancy are most susceptible to adverse effects of vaccination against FMD.

Animals↗

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 contained IgG although the nude mouse serum was deficient in IgG1. Antigen-specific antibody could not be induced in spleen cell cultures from C57BL/6 mice after depletion of T cells with monoclonal antibody plus complement. However, the antibody response could be reconstituted if either a source of exogenous lymphokines or T cells from primed but not unprimed mice were added. Similarly, polyclonal stimulation or unprimed T cells could restore the in vitro response and thus complemented the finding of a low frequency of helper T cells in unprimed mice. Taken together, these data identify that the induction of IgG in FMDV-immunized mice is T cell-dependent and regulated by lymphokines. Furthermore, in nude mice a site other than the spleen must be responsible for the synthesis of the observed serum IgG.

Animals↗

Questionnaire survey of foot and mouth disease (FMD) and of FMD control by vaccination in villages in northern Thailand.

The authors describe a questionnaire survey of foot and mouth disease (FMD) and of FMD control by vaccination in sixty villages in northern Thailand. FMD was found to be common, with 90% of the villages surveyed reporting one or more outbreaks in the five years prior to interview, and 28% of villages reporting outbreaks every year. Outbreaks were more common in the wet season (June to September) and most outbreaks lasted for four weeks or less. The common sources of the most recent outbreak were the introduction of infected cattle and buffalo from a public market or surrounding village (25 of 60 cases), and commingling of cattle and buffalo with those of an infected neighbouring village (24/60). FMD was not perceived to be a major animal health problem, and there was substantial variation between villages in the percentage of animals vaccinated. The main reasons for not vaccinating were the fear that vaccination caused abortion, and the fact that animals were difficult to assemble for vaccination. It was concluded that vaccination cover needs to be raised significantly if FMD is to be effectively controlled.

Abortion, Veterinary↗

Immune response to foot-and-mouth disease virus in a murine experimental model: effective thymus-independent primary and secondary reaction.

The immune response against foot-and-mouth disease virus (FMDV) was studied in a murine model. In untreated control mice, the inoculation of 10,000 suckling mouse 50% lethal doses of Ol Campos FMDV i.p. was followed by a burst of viraemia that disappeared in less than 4 days, i.e. when the neutralizing antibodies (NAb) reached titres above one neutralizing unit. In mice treated with cyclophosphamide, the curves of viraemia and NAb were significantly delayed. Nu/nu mice injected with FMDV had curves of viraemia and NAb identical to those of their nu/t littermates. We then studied the secondary (memory) immune reaction in the same model. In order to investigate which preimmunized cells participate in the elimination of actively replicating FMDV, mice were irradiated, then infected with FMDV, and 24 hr later repopulated with cells obtained from either donor mice that had been previously immunized by infection with live virus, or non-infected controls. The transfer of control (non-immunized) lymphoid cells was unable to eliminate the viraemia in recipient animals at times significantly different from those observed with irradiated recipients receiving no cells, while repopulation of recipients with 10(8) immune lymphoid cells (obtained from pooled thymus, blood, peritoneal exudate, spleen and lymph nodes of preinfected donor mice) led to undetectable titres of viraemia at Day 5 post-infection (p.i.). High doses of thymus cells were totally inactive, while a few as 10(7) donor spleen cells were able to abort viraemia at 6 days p.i. When enriched preparations of B or T spleen cells were adoptively transferred, only B cells were able to abort viraemia in irradiated recipients. It is concluded that, in the murine model of FMDV infection, B cells are mainly responsible for primary response and short-term immunological memory. In both cases the protective immune reaction is T-independent.

Animals↗

Preparation and characterization of monoclonal antibodies to foot-and-mouth disease virus type Asia-1.

Eight clones of hybrid cells secreting monoclonal antibodies to foot-and-mouth disease virus type Asia-1 were prepared; three of them neutralized viral infection. The specificity of THE monoclonal antibodies was analyzed by various immunoenzyme assays using 146S viral particles, trypsin-treated 146S particles, 12S particles, and certain viral polypeptides. The epitopes unique for virus type Asia-1 and conservative among several types were detected on the surface of viral particles. Epitopes on the surface of viral particle were mapped.

Antibodies, Monoclonal↗

Use of peptides for immunization against foot-and-mouth disease.

A peptide corresponding to the major immunogenic site of the protein VP1 of foot-and-mouth disease virus (FMDV) will elicit a protective neutralizing antibody response in guinea-pigs, cattle and pigs. The response is much greater when the peptide is presented as a linear dimer or tetramer and pigs receiving as little as 40 micrograms peptide have been protected against challenge infection. An even greater response is obtained when the peptide is presented as part of the core protein of hepatitis B virus. Moreover, responsiveness to the peptide in non-responder mice can be stimulated by the simultaneous inoculation of an appropriate T-cell epitope linked to the FMDV peptide.

Animals↗

Diagnosis of foot-and-mouth disease by RT-PCR: use of phylogenetic data to evaluate primers for the typing of viral RNA in clinical samples.

The results of type-specific RT-PCR diagnostic assays on foot-and-mouth disease (FMD) viruses in clinical samples were mapped onto serotype-specific dendrograms representing the degree of nucleotide sequence variation between the FMD virus isolates. This novel approach assisted the selection of suitable PCR primer sets for the diagnosis of FMD virus isolates belonging to different topotypes within each serotype. These interpretations were qualified by using a universal (FMD virus group) specific primer to confirm that FMD virus RNA had been extracted from the samples under investigation. The analyses showed that the design of primer sets for the detection of FMD virus serotypes O, A, Asia 1, SAT 1 and SAT 3 were generally satisfactory, as most virus isolates within the major virus sub-groupings were successfully detected. However, the FMD virus serotype C and SAT 2 specific primers were less efficient as certain virus sub-groups were not detected. This identified the need for additional or alternative primers to improve RT-PCR procedures for more comprehensive detection of divergent virus strains within these serotypes. There were some examples where not all virus isolates from the same outbreak reacted with particular type-specific primers which suggested that either further minor refinements may be necessary in the primer design or that there were shortcomings in the RT-PCR methodology.

Animals↗

Virulence of temperature-sensitive mutants of foot-and-mouth disease virus.

A number of temperature-sensitive mutants isolated from two strains of foot-and-mouth disease virus were examined for their virulence in suckling mice. The majority of the mutants were found to be less virulent than the parent virus strains, ranging from slight to total attenuation, but two mutants retained parental levels of virulence. There was no correlation between mutant cut-off temperatures and virulence, or the revertant content of mutant preparations and virulence. It was not always possible to regain parental levels of virulence by isolating phenotypic revertants or wild-type recombinants from genetic crosses, but recombinants were significantly more virulent than either of the two input viruses in a genetic cross.

Animals↗