Pip Hamblin

Foot-and-Mouth Disease Virus Can Induce a Specific and Rapid CD4+ T-Cell-Independent Neutralizing and Isotype Class-Switched Antibody Response in Naïve Cattle

ABSTRACT The role of T-lymphocyte subsets in recovery from foot-and-mouth disease virus (FMDV) infection in calves was investigated by administering subset-specific monoclonal antibodies. The depletion of circulating CD4+ or WC1+ γδ T cells was achieved for a period extending from before challenge to after resolution of viremia and peak clinical signs, whereas CD8+ cell depletion was only partial. The depletion of CD4+ cells was also confirmed by analysis of lymph node biopsy specimens 5 days postchallenge. Depletion with anti-WC1 and anti-CD8 antibodies had no effect on the kinetics of infection, clinical signs, and immune responses following FMDV infection. Three of the four CD4+ T-cell-depleted calves failed to generate an antibody response to the nonstructural polyprotein 3ABC but generated a neutralizing antibody response similar to that in the controls, including rapid isotype switching to immunoglobulin G antibody. We conclude that antibody responses to sites on the surface of the virus capsid are T cell independent, whereas those directed against the nonstructural proteins are T cell dependent. CD4 depletion was found to substantially inhibit antibody responses to the G-H peptide loop VP1135-156 on the viral capsid, indicating that responses to this particular site, which has a more mobile structure than other neutralizing sites on the virus capsid, are T cell dependent. The depletion of CD4+ T cells had no adverse effect on the magnitude or duration of clinical signs or clearance of virus from the circulation. Overall, we conclude that CD4+ T-cell-independent antibody responses play a major role in the resolution of foot-and-mouth disease in cattle.

Emergency vaccination of sheep against foot-and-mouth disease: Significance and detection of subsequent sub-clinical infection

This study has quantified the level of foot-and-mouth disease virus (FMDV) replication and shedding in vaccinated sheep and correlated this to the severity of clinical signs, the induction of antibodies against FMDV non-structural proteins (NSPs) and the transmission of virus to in-contact vaccinated sentinel sheep. To mimic an emergency vaccination regime in the field, sheep were vaccinated with O(1) Manisa vaccine and 4 or 10 days later were indirectly challenged with aerosols from O(1) UKG FMDV infected pigs. Vaccinated and control unvaccinated sheep were monitored for a minimum of 39 days post-challenge. The vaccinated sheep became sub-clinically infected, with reduced virus replication and excretion compared to unvaccinated and clinically infected sheep. Seroconversion to NSP was weak and transient in sheep in which virus replication was of low level and short duration. Virus transmission from vaccinated sub-clinically infected sheep to introduced vaccinated sentinels was not sufficient to cause NSP seroconversion or significant virus shedding. 10% of 10 days and 20% of 4 days vaccinated sheep were virus carriers at greater than 28 days post-challenge compared to 37.5% in the unvaccinated and clinically infected sheep. These results suggest that the low levels of virus replication likely if an effective vaccine is administered at least 4 days prior to challenge exposure are unlikely to result in the spread of infection even under intensive management conditions. Although it may be difficult to detect this infection by serosurveillance, the significance of missing it is likely to be low and the main value of such testing will be to detect undisclosed clinical infection resulting from lack of observation or from exposure to virus before or very soon after vaccination or from vaccine failure due to maladministration or inappropriate strain selection.

Serological Survey for Foot-and-Mouth Disease Virus in Wildlife in Eastern Africa and Estimation of Test Parameters of a Nonstructural Protein Enzyme-Linked Immunosorbent Assay for Buffalo

ABSTRACT In this study we estimate the seroprevalence of foot-and-mouth disease virus (FMDV) in wildlife from eastern and central Africa. Sera were sourced from between 1994 and 2002 from a rinderpest surveillance program. Our study compared a nonstructural protein enzyme-linked immunosorbent assay (Cedi test) with a virus neutralization test. The study shows that there is only a low seroprevalence of FMDV in sampled nonbuffalo species. The seroprevalence in the Cape buffalo was high for SAT2, lower for SAT1, and lowest for SAT3. As the SAT2 serotype was most prevalent, the Cedi test largely reflected the occurrence of SAT2-positive animals. The results also suggest that SAT2 became dominant around 1998, with a large increase in seroprevalence. The sensitivity and specificity of the Cedi test were estimated by comparison to the combined virus neutralization test results from all three SAT tests. A Bayesian implementation of the Hui-Walter latent class model was used to estimate the test parameters. The model permits estimation in the absence of a gold standard test. The final model, using noninformative priors and assuming conditional independence of test performance, estimated Cedi test sensitivity at 87.7% and specificity at 87.3%. These estimates are similar to those for domestic bovines; they suggest that the Cedi test is a useful tool for screening buffalo for infection with the various serotypes of FMDV.

The field effectiveness of routine and emergency vaccination with an inactivated vaccine against foot and mouth disease

High potency, inactivated foot and mouth disease (FMD) vaccines may be used in non endemic countries for emergency vaccination during outbreaks in order to prevent virus spread. In endemic countries either standard or high potency vaccines are used for routine vaccination. Despite their wide use there is a shortage of data on the field effectiveness of inactivated FMD vaccines. Epidemics of FMD caused by viruses of serotype O occur frequently in Israel, where a high potency (≥6PD(50)) vaccine is used for both routine and emergency vaccination. We investigated an outbreak of FMD caused by a virus of serotype O, which took place during 2011 in a feedlot and an adjacent dairy herd. Post outbreak testing of antibodies against non-structural protein demonstrated that infection occurred in 96% of the calves that received two doses of vaccine at least three months prior to the outbreak and more than 50% showed clinical signs consistent with FMD. Replacement heifers that had been vaccinated 3-5 times with the last vaccination administered 7 months prior to the outbreak were all infected and 18% showed clinical signs. Testing of cattle sera of the same vaccination status as the affected cattle demonstrated low neutralizing antibody (NA) titers against the field virus strain and an r(1) value of 0.37 compared to the vaccine strain. In contrast, cattle vaccinated only once but up to two weeks before the outbreak, were almost all protected from clinical disease and to a lesser extent, protected from FMD virus infection, despite low NA titers. We conclude that emergency vaccination was highly effective due to a mechanism not associated with NA, whereas routine vaccination with the same vaccine formulation provided only limited protection due to poor longevity of the elicited immunity and low matching with the field strain (despite an r(1) higher than 0.3).

Interferon-γ Induced by In Vitro Re-Stimulation of CD4+ T-Cells Correlates with In Vivo FMD Vaccine Induced Protection of Cattle against Disease and Persistent Infection

The immune defense against FMDV has been correlated to the antibody mediated component. However, there are occasions when some animals with high virus neutralising (VN) antibody are not protected following challenge and some with low neutralising antibody which do not succumb to disease. The importance of cell mediated immunity in clinical protection is less clear and so we investigated the source and production of interferon-gamma (IFN-γ) in re-stimulated whole blood of FMDV immunized cattle and its correlation to vaccine induced protection and FMDV persistence. We were able to show a positive correlation between IFN-γ response and vaccine induced protection as well as reduction of long term persistence of FMD virus. When combining this IFN-γ response in re-stimulated blood with virus neutralizing antibody titer in serum on the day of challenge, a better correlation of vaccine-induced protection with IFN-γ and VN antibody was predicted. Our investigations also showed that CD4+ T-cells are the major proliferating phenotype and IFN-γ producing cells.

Foot-and-mouth disease virus epitope dominance in the antibody response of vaccinated animals.

Five neutralizing antigenic sites have been identified on the surface of serotype O foot-and-mouth disease virus (FMDV). A set of mAb neutralization-escape mutant viruses was used for the first time to evaluate the relative use of known binding sites by polyclonal antibodies from three target species: cattle, sheep and pigs. Antibodies to all five neutralizing antigenic sites were detected in all three species, with most antibodies directed against antigenic site 2, followed by antigenic site 1. In 76 % of cattle, 65 % of sheep and 58 % of pigs, most antibodies were directed against site 2. Antibodies specific to antigenic sites 3, 4 and 5 were found to be minor constituents in the sera of each of the target species. This implies that antigenic site 2 is a dominant neutralization immunogenic site in serotype O FMDV and may therefore be a good candidate for designing novel vaccines.

Evaluation of laboratory tests for sat serotypes of foot-and-mouth disease virus with specimens collected from convalescent cattle in Zimbabwe

During a field study in Zimbabwe, clinical specimens were collected from 403 cattle in six herds, in which the history of foot-and-mouth disease (fmd) vaccination and infection appeared to be known with some certainty. Five herds had reported outbreaks of disease one to five months previously but clinical fmd had not been observed in the sixth herd. A trivalent vaccine (South African Territories [sat] types 1, 2 and 3) had been used in some of the herds at various times either before and/or after the recent outbreaks of fmd. The primary aim of this study was to evaluate the performance of serological tests for the detection of sat-type fmd virus infection, particularly elisas for antibodies to non-structural proteins (nsps) of fmd virus and solid phase competition elisas (spces) for serotypes sat1 and sat2. Secondary aims were to examine nsp seroconversion rates in cattle that had been exposed to infection and to compare virus detection rates by virus isolation and real-time reverse transcriptase-pcr (rtrt-pcr) tests on both oesophagopharyngeal fluids and nasopharyngeal brush swabbings. In addition, the hooves of sampled animals were examined for growth arrest lines as clinical evidence of fmd convalescence. Laboratory tests provided evidence of fmd virus infection in all six herds; sat2 viruses were isolated from oesophagopharyngeal fluids collected from two herds in northern Zimbabwe, and sat1 viruses were isolated from three herds in southern Zimbabwe. Optimised rtrt-pcr was more sensitive than virus isolation at detecting fmd virus persistence and when the results of the two methods were combined for oesophagopharyngeal fluids, between 12 and 35 per cent of the cattle sampled in the convalescent herds were deemed to be carriers. In contrast, nasopharyngeal swabs yielded only two virus-positive specimens. The overall seroprevalence in the five affected herds varied with the different nsps from 56 per cent to 75 per cent, compared with 81 per cent and 91 per cent by homologous spce and virus neutralisation tests respectively. However, if serological test results were considered only for the cattle in which persistent infection with fmd virus had been demonstrated, 70 to 90 per cent scored seropositive in the different nsps.

Investigations into the cause of foot-and-mouth disease virus seropositive small ruminants in Cyprus during 2007.

In 2007, serological evidence for foot-and-mouth disease (FMD) infection was found as a result of differential diagnostic testing of Cypriot sheep suspected to be infected with bluetongue or contagious ecthyma. Seropositive sheep and goats were subsequently uncovered on ten geographically clustered flocks, while cattle and pigs in neighbouring herds were all seronegative. These antibodies were specific for serotype-O FMD virus, reacting with both structural and non-structural (NS) FMD viral proteins. However, no FMD virus could be recovered from the seropositive flocks. FMD had not been recorded in Cyprus since 1964 and there has been no vaccination programme since 1984. Since all the seropositive animals were at least 3 years old and home-bred, it was concluded that infection had occurred approximately 3 years previously had passed un-noticed and died out spontaneously. It therefore appears that antibodies to FMD virus NS proteins can still be detected around 3 years after infection of small ruminants, but that virus carriers cannot be detected at this time. This unusual situation of finding evidence of historical infection in a FMD-free country caused considerable disruption and alarm and posed questions about the definition of what constitutes a FMD outbreak.

Bovine Serum Panel for Evaluating Foot-and-Mouth Disease Virus Nonstructural Protein Antibody Tests

A panel of 36 sera has been assembled from experimental cattle that had been infected by inoculation or contact exposure with 4 serotypes of foot-and-mouth disease virus (FMDV) with or without prior vaccination. Virus replication and persistence had been characterized in all of the animals. The proportion of the sera scored positive by 5 tests for antibodies to the nonstructural proteins of FMDV varied, suggesting that the panel can discriminate between the sensitivity with which such tests are able to identify infected cattle. Use of this panel will help in assessment of new tests and quality control of existing methods.

Longevity of protection in cattle following immunisation with emergency FMD A22 serotype vaccine from the UK strategic reserve

To determine the longevity of protective immunity following a single administration of emergency vaccine, and establish whether the immune response could be enhanced by increasing the antigen payload even further, cattle were vaccinated with an A22 Iraq vaccine containing either 1x antigen payload (field dose) or 5x antigen payload. Six months post-immunisation all cattle received a homologous virus challenge. The magnitude of the virus neutralising antibody response elicited was consistent with the response to similarly formulated A serotype vaccines with a PD(50) greater than 32. All the vaccinated cattle, regardless of antigen payload, were protected from clinical disease following challenge although some cattle in both groups became sub-clinically infected. We conclude that immunisation with a single inoculation of vaccine from the UK emergency reserve can protect cattle from clinical disease for at least 6 months post-vaccination and that a boost may be unnecessary in an outbreak situation. Some animals may become sub-clinically infected but this is likely to be dependent on the severity of challenge. The study confirmed that a booster at 21 days post-vaccination was not necessary to maintain a cell-mediated response in cattle for 6 months. No increased benefits were recognised by increasing the antigen payload of this vaccine 5x.