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PubMed2006

Interaction of foot-and-mouth disease virus with dendritic cells.

Despite several decades of investigation, the manner in which foot-and-mouth disease virus (FMDV) interacts with the innate and adaptive immune compartments is not completely understood. The importance of elucidating this relationship is emphasized by the inability of current FMDV vaccines to provide long-term protection and the recent outbreaks of FMDV in formerly disease-free countries. Dendritic cells (DCs) are professional antigen-presenting cells that have evolved to monitor the environment and provide a link between the innate and adaptive immune systems. Comprehending the cross-talk between DC and FMDV will provide valuable information towards understanding the host response to the virus and will aid in the design of effective tools and vaccines to block virus spread.

Animals↗
PubMed2005

Cowden's syndrome affecting the mouth, gastrointestinal, and central nervous system: a case report and review of the literature.

Cowden's syndrome (CS) is a rare genodermatosis, of autosomal dominant inheritance and variable phenotype, principally characterized by the formation of hamartomas in various organs, including skin, thyroid, breast, brain, and gastrointestinal tract and by the increased risk for the development of malignancy. The present report details the features of a very rare presentation of Cowden's syndrome affecting not only the mouth and gastrointestinal mucosa but also the central nervous system.

Adult↗
PubMed2004

Re-emergence of foot-and-mouth disease in Botswana.

The re-emergence of foot-and-mouth disease (FMD) in Botswana is reported. The disease outbreak occurred in the Matsiloje Extension Area of Francistown veterinary district situated in the northeastern part of the country in an Office International des épízooties (OIE) recognized FMD free zone without vaccination. The disease affected cattle only and did not spillover into sheep and goats resident in the same extension area, as demonstrated by lack of seroconversion to FMD when tested. The virus isolate associated with the outbreak was identified as FMD virus; Southern African Territories (SAT) type SAT-2. The disease outbreak is discussed in relation to FMD outbreaks that have occurred previously within and outside Botswana.

Animals↗
PubMed2005

Microarray-based detection and typing of foot-and-mouth disease virus.

Foot-and-mouth disease virus (FMDV) is the most economically important veterinary pathogen because of its highly infectious nature and the devastating effects the virus has on the livestock industry. Rapid diagnostic methods are needed for detection and typing of FMDV serotypes and differentiation from other viruses causing vesicular diseases. We developed a microarray-based test that uses a FMD DNA chip containing 155 oligonucleotide probes, 35-45 base pair (bp) long, virus-common and serotype-specific, designed from the VP3-VP1-2A region of the genome. A set of two forward primers and one reverse primer were also designed to allow amplification of approximately 1100 bp of target sequences from this region. The amplified target was labelled with Alexa-Fluor 546 dye and applied to the FMD DNA chip. A total of 23 different FMDV strains representing all seven serotypes were detected and typed by the FMD DNA chip. Microarray technology offers a unique capability to identify multiple pathogens in a single chip.

Animals↗
PubMed2006

Foot-and-mouth disease - quantification and size distribution of airborne particles emitted by healthy and infected pigs.

There is strong evidence to suggest that foot-and-mouth disease (FMD) can be transmitted by airborne virus up to many kilometres from a virus source. Atmospheric dispersion models are often used to predict where this disease might spread. This study investigated whether FMD virus (FMDV) aerosol has specific characteristics which need to be taken into consideration in these models. The characteristics and infectiousness of particles emitted by 12 pigs have been studied pre- and post-infection with O UKG 2001 FMDV. Aerosol generated by individual pigs was found log normally distributed in the range 0.015-20.0microm with concentrations between 1000 and 10000cm(-3) at the smallest size and <1cm(-3) above 10microm. No differences in either the total number of particles produced or their size distribution were detected between uninfected and infected pigs. However, a correlation between aerosol concentration and animal activity was found with a more active pig producing significantly greater concentrations than those that were less active. Viable virus was found up to a maximum of 6.3 log TCID(50)/24h/animal. The virus was distributed almost equally across the three size ranges; <3, 3-6 and >6microm. No correlation could be established between the production of virus and animal activity. In general the production of airborne virus closely followed the detection of viraemia in the blood and the presence of clinical symptoms. However, in one instance a pig excreted as much airborne virus as the other animals in the study, but with less virus detected in its blood. The results suggest that there is little merit in including a sophisticated virus release pattern based on physical activity periods or FMDV aerosol size spectrum, together with the appropriate dry deposition calculations, in models used to predict airborne spread of FMD. An estimate of the total daily virus production based on the clinical assessment of disease and virus strain is sufficient as input.

Air Microbiology↗
PubMed2004

Vaccination of pigs two weeks before infection significantly reduces transmission of foot-and-mouth disease virus.

The objective of this study was to investigate whether and at what time interval could vaccination reduce transmission of foot-and-mouth disease virus (FMDV) among pigs. Reduction of virus transmission by vaccination was determined experimentally. Transmission of FMDV was studied in three groups of ten pigs: one non-vaccinated group and two groups that were vaccinated 7 days (-7 dpi) and 14 days before inoculation (-14 dpi), respectively. Five randomly selected pigs from each group were inoculated with FMDV type O Taiwan, while the other five pigs left in the groups were exposed to the inoculated pigs by direct contact. Clinical signs were recorded, virus isolation and RT-PCR were carried out on oropharyngeal fluid (OPF), and the neutralizing antibody titres and the antibody response against non-structural (NS) proteins of FMDV were determined. No virus transmission was observed in the -14 dpi group, whereas virus transmission was observed in all contact pigs affecting both the non-vaccinated and the -7 dpi group. The reproduction ratio R in the -14 dpi vaccinated group was significantly lower than that of the non-vaccinated group. This study confirms the potential of vaccination as an important tool to reduce transmission of FMDV.

Animals↗
PubMed2004

Reintroduction of foot-and-mouth disease in Argentina: characterisation of the isolates and development of tools for the control and eradication of the disease.

This paper describes the antigenic and molecular characterisation of foot-and-mouth disease virus (FMDV) strains isolated during the 2000-2002 epidemic in Argentina, and the strategy implemented for disease control. Two different FMDV serotypes, O and A, were involved. Of the various field isolates studied, two distinct O1 lineages (strains Corrientes/00 and Misiones/00) and two serotype A lineages (A/Argentina/00 and A/Argentina/01 prototypes) were identified. The genome sequences of these strains were compared with sequences of previous regional isolates and sequences of vaccine strains. O1 strains were found to be related to regional strains while serotype A strains were found to be more distanced from them. The updating of the antigenic composition of the vaccines used in the emergency was a key issue, since the outbreaks stopped shortly after the implementation of the vaccination programs. The O1 strains quickly disappeared from the field following strict control measures and the use of vaccines containing O1/Campos strain. However, in the case of the A serotype strains, the situation was different, since the use of a vaccine containing strain A24/Cruzeiro yielded acceptable levels of protection only after re-vaccination. Therefore, the new field strains A/Argentina/00 and A/Argentina/01 were incorporated into the vaccine, leading to an effective control of the disease. Viral circulation greatly diminished, as indicated by the significant reduction in the number of outbreaks and in the number of animals with antibodies against non-structural proteins. Satisfactory levels of protective antibodies were subsequently detected in the cattle population (above 75% protection). The absence of outbreaks after January 2002 indicated that the epidemic was controlled.

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↗
PubMed2005

Development of transgenic alfalfa plants containing the foot and mouth disease virus structural polyprotein gene P1 and its utilization as an experimental immunogen.

The use of transgenic plants as vectors for the expression of viral and bacterial antigens has been increasingly tested as an alternative methodology for the production of experimental vaccines. Here, we report the production of transgenic alfalfa plants containing the genes encoding the polyprotein P1 and the protease 3C of foot and mouth disease virus (FMDV). The immunogenicity of the expressed products was tested using a mouse experimental model. Parenterally immunized mice developed a strong antibody response and were completely protected when challenged with the virulent virus. This report demonstrates the possibility of using transgenic plants to express polyprotein P1 and the protease 3C of FMDV and their utilization as effective experimental immunogens.

Agrobacterium tumefaciens↗
PubMed2005

The application of new techniques to the improved detection of persistently infected cattle after vaccination and contact exposure to foot-and-mouth disease.

Detection of antibodies to the non-structural proteins (NSP) of foot-and-mouth disease virus (FMDV) was compared with conventional serological and virological methods and with RT-PCR for the identification of FMDV carrier animals obtained after experimental contact challenge of vaccinated cattle. Transmission from carriers to sentinels was also monitored. Twenty FMDV vaccinated and five unvaccinated cattle were challenged by direct contact with five donor cattle excreting FMDV and monitored until 28 days post challenge-exposure . Twelve vaccinated and three unvaccinated animals were retained up to 24 weeks post exposure to FMDV in order to monitor viral persistence, transmission and antibody responses. In nine vaccinated animals, infection persisted beyond 28 days post exposure, virus being detected more frequently and for longer in oesophagopharyngeal samples from these animals when examined by RT-PCR rather than by virus isolation. Although recovery of FMDV RNA became increasingly sporadic over time, the number of RNA copies detected in positive samples declined only slowly. Two naïve sentinel cattle housed with the persistently infected animals between 93 and 168 days after the latter had been challenge-exposed to FMDV did not become infected. There were differences in the ability of commercially available serological tests to detect antibodies to FMDV non-structural proteins (NSP) in vaccinated and subsequently challenged cattle. Although no single test could identify all of the vaccinated cattle that became persistently infected, the most poorly recognised animals were those with the least evidence of virus replication based on other tests. The potential of the detection of antibodies to the 2B NSP of FMDV for diagnosing persistent FMDV infection was demonstrated.

Animals↗
PubMed2005

Expressions of Bovine IFN-gamma and foot-and-mouth disease VP1 antigen in P. pastoris and their effects on mouse immune response to FMD antigens.

As a highly contagious disease in cloven-hoofed animals, foot-and-mouth disease virus (FMDV) may cause a considerable social-economic loss in those countries affected. IFN-gamma has a wide range of antiviral and immune modulating functions. Thus, to study the immune enhancing effects of recombinant Bovine IFN-gamma (rBoIFN-gamma) on a recombinant FMDV vaccine, BoIFN-gamma, FMDV VP1 and BoIFN-gamma/VP fusion genes were cloned, expressed, co-expressed in pichia pastoris (P. pastoris) respectively, and subsequent immune effects have been evaluated in this study. The results showed that the genes encoding for BoIFN-gamma, VP1 and BoIFN-gamma/VP1 are successfully expressed in P. pastoris and their products are directly secreted into the cultural supernatant at a high level of 1.0 g/L analyzed by thin-layer scanning. In addition, rVP1 alone could induce both humoral and marginal cell-mediated immune responses in mice, while the group with co-inoculations of rBoIFN-gamma could markedly enhance both humoral and cell-mediated immune responses; even more dramatic immune responses were observed with the group inoculated with the fusion product, rBoIFN-gamma/VP1. The fusion product could be further investigated for its utility of FMDV vaccine development.

Animals↗
PubMed2005

Interferon-gamma production in vitro from whole blood of foot-and-mouth disease virus (FMDV) vaccinated and infected cattle after incubation with inactivated FMDV.

Studies were performed to determine whether a rapid method to detect cell mediated immune responses to foot-and-mouth disease virus (FMDV) could be used either as a diagnostic test or provide a correlate of protection in animals post-vaccination. Using protocols based on the BOVIGAM assay for tuberculosis, whole blood samples from FMDV vaccinated or control animals, before and after challenge infection, were stimulated overnight with inactivated FMDV antigen. The quantity of interferon gamma (IFN-gamma) produced in the supernatants was measured using an ELISA. Specific induction of IFN-gamma was detected in samples from vaccinated, infected and vaccinated-then-infected cattle. Further development of this assay may provide a useful tool for the diagnosis of FMDV immune animals, including the identification of vaccinated animals that have been subsequently infected with FMDV. In these studies, combining the results of the IFN-gamma assay with virus neutralising antibody titre, in groups of vaccinated animals, provided a correlation with the capacity to control virus replication after subsequent challenge.

Animals↗
PubMed2005

Comparison of immune responses after intra-typic heterologous and homologous vaccination against foot-and-mouth disease virus infection in pigs.

This study compares the immune responses and protection induced by intra-typic heterologous vaccination with that induced by homologous vaccination against challenge with foot-and-mouth disease virus (FMDV). Humoral and cell-mediated immune responses and protection against challenge with FMDV O Taiwan were examined in a non-vaccinated group, a group vaccinated with O Taiwan FMD vaccine and a group vaccinated with O Manisa FMD vaccine. Five pigs from each group were challenged with FMDV type O Taiwan 14 days after vaccination and five other pigs were contact-exposed to the inoculated pigs. Both homologous and heterologous vaccination protected against challenge with FMDV O Taiwan at 2 weeks after vaccination. In the heterologous vaccinated group, cross-neutralizing antibody titres against O Taiwan could be detected although the ratio 'r(1)' was 0.4, which was significantly smaller than the critical r-value. Cell-mediated immune responses were detected after both homologous and heterologous vaccination. Virus-induced in vitro lymphocyte (cross-) proliferation and production of both a Th1-type (IFN-gamma) and a Th2-type (IL-10) cytokine response were demonstrated in cultures of peripheral blood mononuclear cells (PBMC). The findings show that heterologous (emergency) vaccination can prevent clinical disease and shedding of virus. The induction of the cell-mediated immune responses after (heterologous) vaccination needs more research but data on these responses might provide additional tools for both vaccine choice and vaccine development.

Animals↗
PubMed2005

Simultaneous detection of antibodies to foot-and-mouth disease non-structural proteins 3ABC, 3D, 3A and 3B by a multiplexed Luminex assay to differentiate infected from vaccinated cattle.

For the first time, a multiplex bead immunoassay was used to test simultaneously, with a single sample, the immune response to foot-and-mouth disease non-structural proteins 3ABC, 3A, 3B and 3D from experimentally infected and vaccinated cattle. We cloned and expressed these non-structural proteins (NSPs) as recombinant antigens. The purified proteins were coupled to microspheres labeled with anti-His monoclonal antibody with different proportions of red and orange fluorescent dyes and reacted against serum specimens. Antibody reacting against different NSPs, and thus, the different colored beads, was detected by use of the Luminex system. This multiplex bead immunoassay can detect the immune response to NSPs in cattle as early as 7 days post-infection. In general antibodies to the protein 3D appeared early after infection and anti-3ABC antibodies were detected at higher levels than the other NSPs. A clear differentiation was established between infected and vaccinated or uninfected cattle. The multiplex bead immunoassay was compared to individual indirect enzyme-linked immunosorbent assays (iELISAs) for the same NSP's responses. Results indicated that this new assay had a high positive correlation with those generated by iELISA. The Luminex-based technology promises to be a sensitive and efficient method that permits multiplexed NSP antibody detection from a single sample and would therefore provide both a time and cost saving to the laboratory.

Animals↗
PubMed2005

Enhanced immunogenicity of microencapsulated multiepitope DNA vaccine encoding T and B cell epitopes of foot-and-mouth disease virus in mice.

The role of poly(D,L-lactic-co-glycolic acid, PLGA) microparticles on enhancing immune responses of multiepitope DNA vaccines was investigated in vitro and in vivo. pcDNA-SG encoding T and B cell epitopes of foot-and-mouth disease virus (FMDV) was encapsulated into PLGA microparticles. PLGA microparticles could protect themselves from nuclease degradation in vitro. PLGA-pcDNA-SG microparticles could be uptaken by cells and expressed His-tagged SG immunogen in vitro and in vivo. A prolonged expression and presentation of SG immunogen were observed by confocal laser scanning microscopy in the lymphocytes from the mice incubated with PLGA-pcDNA-SG microparticles, compared with the mice immunized with naked pcDNA-SG. PLGA-pcDNA-SG microparticles displayed a significant stronger immunogenicity than naked DNA vaccines with a higher titer of virus-specific antibody, elevated IFN-gamma production and enhanced lymphocyte proliferation. PLGA-DNA microparticle could elicit augmented humoral and cellular responses with reduced amounts and times of immunization.

Animals↗
PubMed2006

The effect of vaccination on foot and mouth disease virus transmission among dairy cows.

The aim of this study was to quantify the effect of a single vaccination of dairy cows on foot and mouth disease virus (FMDV) transmission. To estimate if vaccination could significantly reduce virus transmission, we performed two replicates of a transmission experiment with one group of vaccinated and one group of non-vaccinated dairy cows (ten animals per group). Half of both groups were intranasally inoculated, with FMDV field isolate O/NET2001, and housed with the other half of the group (contact-exposed cows) from the next day onwards. Virus transmission was quantified by estimating the reproduction ratio R, which is the average number of secondary cases caused by one infectious animal. In the non-vaccinated groups all cows became infected and Rnv was significantly above 1. In the vaccinated groups infection was demonstrated in three inoculated cows, and no transmission was observed (Rv was 0, not significantly below 1). Transmission was significantly reduced in the groups of vaccinated cows when compared to the groups of unvaccinated cows. Our findings indicate that after a single vaccination cows are protected against infection of FMD and that most likely no virus transmission will occur within a vaccinated herd.

Animals↗
PubMed2005

Extract from Agaricus blazei Murill can enhance immune responses elicited by DNA vaccine against foot-and-mouth disease.

The fungus Agaricus blazei Murill (ABM) is particularly rich in polysaccharides, which have shown particularly strong results in treating and preventing cancers. The goal of this study was to investigate whether co-administration of the ABM extract with foot-and-mouth disease virus (FMDV) DNA vaccine could increase the immune responses. Compared with the control mice, which received FMDV DNA vaccine alone, significant increase in not only the FMDV-specific antibody response but also T cell proliferation was observed in mice which received FMDV DNA vaccine plus the ABM extract. Taken together, these results demonstrated that application of the ABM extract might provide a strategy to improve the efficacy of DNA vaccines.

Adjuvants, Immunologic↗
PubMed2004

Comparable sensitivity and specificity in three commercially available ELISAs to differentiate between cattle infected with or vaccinated against foot-and-mouth disease virus.

Three commercially available ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus (FMDV) were evaluated, using sera from uninfected, vaccinated, infected, inoculated, first vaccinated and subsequently infected, and first vaccinated and subsequently inoculated cattle. We compared antibody kinetics to non-structural proteins, sensitivity, and specificity. One of the ELISAs had a higher sensitivity and much lower specificity than the other two, therefore we established standardised cutoff values for the compared assays using receiver operated characteristic (ROC) curves. Using the standardised cutoff values, all three ELISAs produced comparable results with respect to sensitivity and specificity. Antibody development to non-structural proteins after infection and after vaccination/infection was not significantly different. Development of antibodies, however, both neutralising and directed to non-structural proteins, was significantly delayed after intranasal inoculation as compared to intradermolingual infection. Based on results of sera obtained after vaccination and experimental infection all three assays can be used for testing sera collected between 4 weeks and 6 months after infection. More information is needed on the prevalence of positive reactors in a situation where emergency vaccination has been used and FMD transmission was still observed.

Animals↗