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PubMed2004

Comparative immunogenecity of foot and mouth disease virus antigens in FMD-haemorrhagic septicaemia combined vaccine and FMD vaccine alone in buffalo calves.

Humoral immune response was evaluated by monitoring the serum antibody titres and virus specific IgM titres against Foot and Mouth Disease (FMD) virus antigens in serum samples obtained from different groups of calves inoculated with combined vaccine or FMD vaccine alone, on 0, 7, 14, 21, 28, 42 and 56 days post-vaccination (DPV). The cellular immune response was monitored by MTT based lymphoproliferation in peripheral blood mononuclear cell cultures. Higher liquid phase blocking (LPB) ELISA antibody titres were observed in calves receiving combined vaccine as compared to calves immunized with FMD vaccine alone with the peak titres in both the groups obtained on 21 days post-vaccination. However, the virus specific IgM titres were significantly higher in group of calves inoculated with combined vaccine than FMD vaccine alone. The lymphoproliferative responses against FMDV types O, A22 and Asia 1 in the groups receiving combined vaccine and FMD vaccine alone started increasing gradually after day 14 and reached peak levels on 28 DPV followed by a gradual decline subsequently. The group receiving combined vaccine showed higher proliferative responses on in vitro stimulation with FMD virus type O, whereas, with FMD virus type Asia 1, the responses were significantly higher on 14 and 21 DPV as compared to the group immunized with FMD vaccine alone. However, in the group receiving combined vaccine, the responses on in vitro stimulation with FMD virus type A22 were significantly higher than FMD vaccine alone group on all DPV except on 42 DPV.

Animals↗
PubMed2005

Protective immune response against foot-and-mouth disease virus challenge in guinea pigs vaccinated with recombinant P1 polyprotein expressed in Pichia pastoris.

Vaccination of the susceptible livestock with potent, safe and cost effective vaccine is the primary requirement to control foot-and-mouth disease (FMD) in an endemic country. In this study, an alternative approach was used in which structural protein genes of all the four serotypes of FMDV (O, Asia 1, A22 and C) were expressed separately in methylotrophic yeast Pichia pastoris. The recombinant polyproteins (P1) were characterized by SDS-PAGE and in Western Blot analysis. Partially purified protein was used for immunization in guinea pigs with different adjuvant formulations and immune response studied. Ninety micrograms of the recombinant protein per monovalent dose was used for immunization. A single injection of a monovalent or polyvalent vaccine was given to guinea pigs with various adjuvant combinations viz., Monovalent recombinant protein either adjuvanted with Montanide-ISA50V or Indigenous oil, Monovalent recombinant protein mixed with 1/10th dose of inactivated oil-adjuvanted virus vaccine and Polyvalent recombinant protein with Montanide ISA50V. FMDV specific humoral immune response was observed at about 28th day post vaccination. The immune response as assessed by indirect ELISA and Serum neutralization test titres was found to be 320-640 and 16-32, respectively. When challenged with virulent homologous type 'O' virus, the guinea pigs showed protective C index of 2.01,1.81, 2.56 and 2.48, respectively, with above said adjuvant combinations. The study has shown that yeast-expressed FMDV P1 polyprotein in a single dose could elicit a protective immune response in guinea pigs, and this could be a possible future vaccine candidate in homologous host.

Animals↗
PubMed1983

Antibody response in pig nasal fluid and serum following foot-and-mouth disease infection or vaccination.

Nasal fluid and serum collected from pigs after exposure to live foot and mouth disease (FMD) virus or injection of single oil emulsion (w/o) or double oil emulsion (w/o/w) vaccines were examined for FMD neutralizing activity. After virus exposure the response profiles of serum and nasal mucus were similar to one another. In both, neutralizing activity rose to a peak at one to two weeks after exposure and then subsided slowly. After vaccination with either the w/o or w/o/w preparations a neutralizing response was demonstrable in the serum three to seven days after the first injection, and this was boosted by revaccinations 56 and 117 days later. The neutralizing activity was also detectable in nasal fluid seven days after the first vaccination, but subsequent revaccinations 56 and 117 days later provoked neutralizing titres which were no greater than those observed after the initial vaccination.

Animals↗
PubMed2000

Genetic and antigenic variance of foot-and-mouth disease virus type Asia1.

The capsid protein encoding genes of five recent type Asia1 foot-and-mouth disease virus isolates, representative of three genotypes, were sequenced. The deduced amino acid sequences were aligned to each other and to two published sequences. The sequence differences suggested different antigenic properties of the isolates. One isolate was used to generate monoclonal antibodies (mAbs) which were analyzed for neutralizing activity and reactivity with trypsinized virus. Trypsin removes the major antigenic sites located at VP1. The five virus isolates formed three reaction patterns with the mAbs, irrespective of their genotype. Combination of all data allowed to suggest the location of the epitope of each antibody: the VP1 G-H and the VP2 B-C loop, the VP3 B-B knob, and the N-terminus of VP2, respectively, were involved.

Amino Acid Sequence↗
PubMed2006

Sequence analysis of the non-structural 3A and 3C protein-coding regions of foot-and-mouth disease virus serotype Asia1 field isolates from an endemic country.

A total of 18 foot-and-mouth disease virus (FMDV) serotype Asia1 field isolates belonging to two different lineages (including the divergent group) as delineated earlier in VP1-based phylogeny were sequenced in the non-structural 3A and 3C protein-coding regions. The phylogenetic trees representing the regions coding for the non-structural proteins were very similar to that of the structural VP1 protein-coding region. Phylogenetic comparison at 3C region revealed clustering of Asia1 viruses with the isolates of serotypes O, A and C in the previously identified clade. Comparison of amino acid sequences identified lineage-specific signature residues in both the non-structural proteins. Overall analysis of the amino acid substitutions revealed that the 3A coding region was more prone to amino acid alterations than 3C region.

Animals↗
PubMed2002

Evidence of recombination in the capsid-coding region of type A foot-and-mouth disease virus.

Recombination is one of the factors that contribute to genetic diversity in foot-and-mouth disease virus (FMDV). Similarity and bootscan analyses have provided evidence of recombination in the capsid-coding (P1) region of the virus. In the present study, of the 14 subtype A22 field isolates that were distributed in three previously described genotypes (IV, VI and VII) based on the 1D (VP1-encoding) gene sequence (Tosh et al., 2002 ), one isolate (IND 170/88) was found to be a hybrid of genotypes VI and VII in the P1 region. VP1, VP4, the 5' region of VP2 and the 3' region of VP3 of this virus were characteristic of genotype VI, whereas the remaining 3' region of VP2 and the 5' region of VP3 were characteristic of genotype VII. No insertion or deletion was observed in the recombinant virus. Recombination in the P1 region may provide an escape mechanism for the virus.

Animals↗
PubMed1999

Serotyping of foot-and-mouth disease virus by antigen capture reverse transcriptase/polymerase chain reaction.

The technique of capturing of foot-and-mouth disease virus (FMDV) from clinical material in microcentrifuge tubes coated with type-specific antibodies and amplifying the viral sequences by RT/PCR in the same tube, promoted the detection and serotyping of FMDV with high sensitivity and specificity. The efficiency of antigen capturing and shelf life of the coated tubes was improved by glutaraldehyde fixation of antibodies to the tubes. Virus in infected tissues, even after storage for 25-30 years at 70 degrees C, could be successfully typed by this method. Conserved sequences flanking the variable region of immunoreactive VP1 gene of FMDV were used as primers in the assay and hence the nucleotide sequence analysis of the product could reveal the strain variation. The test has been found to be at least 125-fold more sensitive than type specific ELISA and of comparable sensitivity as other protocols for detection of FMDV by RT/PCR.

Animals↗
PubMed2006

Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus.

To validate the use of serology in substantiating freedom from infection after foot-and-mouth disease (FMD) outbreaks have been controlled by measures that include vaccination, 3551 sera were tested with six assays that detect antibodies to the non-structural proteins of FMD virus. The sera came from naïve, vaccinated, infected and vaccinated-and-infected animals; two-thirds from cattle, the remainder from sheep and pigs. The assays were covariant for sensitivity, but not necessarily for specificity. A commercial kit from Cedi-diagnostics and an in-house assay from IZS-Brescia were comparable to the NCPanaftosa-screening index method described in the Diagnostic Manual of the World Animal Health Organisation. Using these three tests the specificity and sensitivity for the detection of carriers in vaccinated cattle approaches or exceeds 99% and 90%, respectively.

Animals↗
PubMed2005

[High expression of the foot-and-mouth disease's structural protein P1 in Escherichia coli and analysis of its biology activity].

Foot-and-mouth disease virus (FMDV) is the aetiological angent of a highly contagious viral disease. The complete gene encoding the structural protein of FMDV (P1) was subcloned into expression vector pGEX-KG, resulting in the fusion expression plasmid pKG-P1. After transformed into E. coli BL21(DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-P1 fusion protein was expressed in high level. The molecular weight of the fusion protein wa 110kD and the expressed products were soluble. Western-blotting was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV. The fusion proteins were further purified by GST purification kit and an indirect ELISA (P1-ELISA) based on the purified proteins was developed. Comparison between P1-ELISA and the standard indirect haemagglutinin assay showed the two methods had 87 per cent agreement by detecting 864 serum samples, indicating the purified P1 protein was specific as the antigen of indirect P1-ELISA.

Capsid Proteins↗
PubMed1988

A comparison of enzyme-linked immunosorbent assay, complement fixation and virus isolation for foot and mouth disease diagnosis.

A total of 205 epithelial tissue samples were examined for the presence of foot and mouth disease virus by either the complement-fixation (CF) test, enzyme-linked immunosorbent assay (ELISA) and/or by virus isolation in bovine thyroid or kidney cell cultures. The virus was isolated from 134 of the 201 (67%) specimens. Samples, from which virus was isolated, were termed virus-positive samples. The CF test detected viral antigen in 30 (24%) of 123 virus-positive samples, whereas the ELISA detected it in 100 (81%) of these specimens. The ELISA was thus at least 3 times more efficient than the CF test in detecting the virus in epithelial-tissue samples. There were 5 samples from which virus was not isolated but which were positive with the ELISA procedure. The ELISA was particularly useful for testing samples from pigs and for assessing specimens from animals with resolving lesions. The ELISA gave virus type-specific results in 89% of 63 virus-positive cases compared to 40% for the CF test. The ELISA was thus a very useful, accurate and sensitive method for the direct testing of epithelial tissues of affected animals.

Animals↗
PubMed1995

Direct evaluation of the immunodominance of a major antigenic site of foot-and-mouth disease virus in a natural host.

The immunodominance of a major antigenic site of foot-to-mouth disease virus (FMDV) (serotype C; clone C-S8c1) in a natural host has been evaluated by serum immunoglobulin fractionation. Nineteen sera from either convalescent or vaccinated swine were fractionated by affinity chromatography using a synthetic peptide representing antigenic site A (the G-H loop of capsid protein VP1) coupled to a Sepharose matrix. Antigen-binding and neutralizing activities of serum fractions were quantitated. On average, about 57 or 27% of the virus-neutralizing activity (and about 35 or 12% of the virus-binding activity) from convalescent or vaccinated swine, respectively, corresponded to antibodies against site A. The results provide direct evidence of the important contribution of site A, and also of additional sites unrelated to site A, in the evoking of neutralizing antibodies by FMDV in a natural host. The proportion of antibodies directed to site A varied greatly among individual swine. Some animals evoked remarkably low levels of antibodies specific for site A although they were competent to raise antibodies against other antigenic sites of FMDV. Thus, the major antigenic site of FMDV shows heterogeneous dominance in a natural host. Possible implications for evolution of viral quasispecies are discussed.

Amino Acid Sequence↗
PubMed1999

Chaperonin 10 of Mycobacterium tuberculosis induces a protective immune response to foot-and-mouth disease virus.

Chaperonin 10 of M. tuberculosis conferred partial or total protection against generalized foot-and-mouth disease (FMD) in guinea-pigs challenged with O1 Lausanne FMD virus. Chaperonin 10-immunized animals mounted an antibody response to the protein, one epitope of which was found in the C-terminal half. A similar recognition pattern was observed in FMD-convalescent guinea-pigs, swine and cattle. Anti-chaperonin 10 sera showed antiviral activity against FMDV-infected BHK-21 cells. There was strong evidence that early after infection these cells actively secrete their histones and that antisera to the chaperonin recognize them. The same antisera reacted with purified histones in immunoblotting. Most important, exogenously added histones abrogated the anti-viral activity of the antiserum and an anti-histone monoclonal antibody had strong antiviral activity against FMDV-infected BHK-21 cells. These results are consistent with previous reports on displacement of histones from the nuclear compartment and immune recognition of self-histones after viral infections. On the whole, they indicate that M. tuberculosis chaperonin 10 enables the immune system to react against early abnormalities of virus-infected cells; this is accomplished by antibody cross-reacting with histones released during virus infection.

Amino Acid Sequence↗
PubMed1964

PRODUCTION AND PURIFICATION OF MILLIGRAM AMOUNTS OF FOOT-AND-MOUTH DISEASE VIRUS FROM BABY HAMSTER KIDNEY CELL CULTURES.

A stable line of baby hamster kidney cells for use in the production of, and subsequent purification of, foot-and-mouth disease virus (FMDV) was grown in large quantities on the cylindrical surfaces of 2-liter Baxter bottles. The bottles, in round wire cages, were rotated on a three-tiered roller mill. The cells retained their rapid growth characteristics and susceptibility to FMDV in a tris(hydroxymethyl)aminomethane buffer-containing medium which was especially formulated for large-scale work. This medium, without being changed, sustained cell growth for 6 to 7 days to yield confluent layers containing 500 to 750 million cells per bottle. In small-scale virus-growth experiments, harvested fluids contained about 10(3.8) to 10(8.8) plaque-forming units (PFU) per ml. This corresponded to a yield of 30 to 50 PFU per cell. In production runs with 190 cultures, the infectious fluids usually contained 10(7.9) to 10(9.2) PFU per ml, and the mass of essentially pure virus obtained therefrom ranged from 7 to 17 mg concomitant with cumulative infectivity recoveries of about 20%.

Animals↗
PubMed1998

The foot and mouth disease virus type O outbreak of 1992 is not related to vaccine strain (O/R2/75).

Vaccination is the only pragmatic approach to control foot and mouth disease in India. Strict quality control measures are essential to supply potent vaccine to the field application, in addition to monitoring the performance of the vaccine in the field. During the process of monitoring, an outbreak of FMD in vaccinated animals caused by type "O" virus in Tanjavur district of Tamil Nadu and a type "O" virus from unvaccinated herd of Karnataka were studied. Field isolates and vaccine virus were sequenced and analyzed. Data indicated that the virus from the outbreak in vaccinated cattle was a variant which could escape neutralization by antibodies against vaccine virus.

Amino Acid Sequence↗
PubMed1998

Antibody to the nonstructural proteins of foot-and-mouth disease virus in vaccinated animals exposed to infection.

Cattle which have been infected with foot-and-mouth disease (FMD) virus can be differentiated from those that have been vaccinated on the basis of the detection of antibody to one or more of the non-structural (NS) proteins of the virus. Cattle which have been protected by vaccination can become persistently infected with FMD virus (FMDV) without ever showing clinical signs. Vaccinated, protected cattle which are persistently infected cannot be distinguished from animals that merely have been vaccinated on the basis of serological tests for antibody to the structural proteins of FMDV. Sera were collected from groups of cattle for varying periods after exposure to infection under experimental conditions. On the basis of isolation of virus from probang samples collected during the course of the experiments it was possible to classify the cattle according to the following criteria; naive, infected and eliminated the virus (convalescent), infected and persistently infected with FMDV (carriers), vaccinated alone, vaccinated and either convalescent or carrier. Sera were examined for antibody to the NS proteins Lb, 2C, 3A, 3D, and 3ABC by an indirect profiling ELISA using E. coli-expressed fusion proteins as antigens. Considerable variation was observed in the antibody response to NS proteins of both naive and vaccinated animals following infection. The extent of individual variation was so great that convalescent animals could not be differentiated from carrier animals on the basis of their antibody response to any of the NS proteins examined. The majority of vaccinated, protected animals showed an antibody response to NS proteins, particularly 3ABC, following exposure to infection. However, the carrier state was demonstrated in some vaccinated, protected animals in which no antibody response to any of the NS proteins could be detected. The detection of antibody to NS proteins can therefore be used on a group, or herd, basis to detect circulation of virus in a vaccinated population but further investigations in the field are required to determine the sampling level necessary for statistical acceptance. On an individual animal basis, however, freedom from antibody to NS proteins in a vaccinated animal, or an animal of unknown history, does not necessarily imply that the animal is free from infection with FMD virus and, furthermore, the titre of antibody to NS proteins is not a useful predictive measure of whether or not an infected animal has successfully eliminated the virus.

Animals↗
PubMed2005

Analysis of the immune response against mixotope peptide libraries from a main antigenic site of foot-and-mouth disease virus.

The design of vaccines for RNA viral diseases is complicated by the high genetic variability of the viruses, which favors the selection of escape mutants. A case in point is foot-and-mouth disease virus (FMDV), for which only limited protection has been observed in vaccination with single peptides. We have explored the potential of immunogens of higher sequence diversity, covering a broad range of field or culture-induced mutations at the immunodominant site A of FMDV, serotype C. Four mixotope-type peptide libraries, containing ca. 3 x 10(3) or ca. 3 x 10(5) peptides each, in either linear or cyclic form, and combining most significant mutations found or induced at site A have been synthesized and used to immunize guinea-pigs. Substantial levels of serum conversion have been observed for all four mixotope libraries, as well as for single peptides, linear or cyclic, corresponding to the consensus site A sequence. The specificity and neutralizing ability of the anti-mixotope and -peptide antibodies have been evaluated by direct ELISA and by plaque reduction and micro-neutralization assays, respectively. Challenge experiments with an infectious, guinea-pig-adapted FMDV strain, have shown higher protection rates in animals immunized with the cyclic versions, either in single sequence or in combinatorial mixotope form.

Animals↗
PubMed1983

Aerosol exposure of cattle to foot-and-mouth disease virus.

Slight modifications of a small, plastic covered greenhouse provided a chamber for the exposure of cattle of all ages to aerosols of foot-and-mouth disease virus. Particle size distributions of aerosols were 76% less than 3 microns, 17% 3-6 microns, and 7% greater than 6 microns immediately after the deVilbis no. 40 nebulizer used was turned off and 90% less than 3 microns, 8% 3-6 microns, and 2% greater than 6 microns 20-30 min later. Pharyngeal virus growth curves and viremia patterns correlated well with the dose of virus to which test cattle were exposed and were similar to those of cattle inoculated intranasally.

Aerosols↗
PubMed1971

Effects of oxyamylose and polyacrylic acid on foot-and-mouth disease and hog cholera virus infections.

Two interferon-inducing polycarboxylates were tested for antiviral activity on foot-and-mouth disease (FMD) virus infections in mice, guinea pigs, and swine. Polyacrylic acid, given intraperitoneally, had a protective effect on infection by FMD virus administered in the peritoneal cavity of mice and in the foot pad of guinea pigs. Chlorite-oxidized oxyamylose (COAM) was effective in mice at a dosage of 2 mg/kg. Swine were not protected against naturally transmitted FMD by 120 mg/kg of COAM nor by polyacrylic acid. Swine were not totally unresponsive to COAM since it delayed symptoms of hog cholera. Interferon was not detected in the serum of COAM-treated swine. With FMD virus, an example was found of activity of interferon inducers in experimental hosts and lack of activity in a natural host.

Acrylates↗