A synthetic peptide containing the consensus sequence of the G-H loop region of foot-and-mouth disease virus type-O VP1 and a promiscuous T-helper epitope induces peptide-specific antibodies but fails to protect cattle against viral challenge

A pilot study was carried out in cattle to determine the immunogenicity of a synthetic consensus peptide comprising the G-H loop region of foot-and-mouth disease virus (FMDV) type-O VP1 and a non-VP1 T-helper (Th) epitope. Cattle vaccinated intramuscularly either once (n = 5) or twice (n = 4) with 50 microg of the peptide preparation at a 21-day interval developed antibodies to the peptide as determined by ELISA with the exception of one steer that received a single dose. However, neutralizing antibody titers against FMDV type-O were modest and all animals presented with clinical FMD signs upon challenge 21 days after the last vaccination. In contrast, four of the five animals inoculated with an inactivated FMD type-O commercially prepared vaccine developed neutralizing antibodies and were fully protected against clinical disease following virus challenge 21 days post-vaccination (dpv). Nucleotide sequence comparison of the VP1 region between the challenge virus and RT-PCR products recovered from a lesion of the peptide-vaccinated animal with the highest neutralizing antibody titer 5 days post-challenge (dpc) showed no evidence for selection of a neutralization-resistant mutant. We conclude that although the synthetic peptide induced an antibody response in cattle, it failed to confer protection against FMDV challenge.     

Aspects of heat inactivation of foot-and-mouth disease virus in milk from intramammarily infected susceptible cows

In skim milk obtained from susceptible cows after intramammary and intravenous inoculation (primary infected milk), foot-and-mouth disease (FMD) virus type O1 was slower inactivated by heat treatment than virus that had been added to pre-exposure skim milk. Residual virus infectivity in heated primary infected milk was more efficiently detected in bovine thyroid cell cultures than in secondary pig kidney (PK2) cell cultures. Untreated primary infected milk was found to inhibit both FMD-virus and vesicular stomatitis virus plaque formation in PK2 cells, suggesting the presence of interferon. The results of further tests confirmed that the interfering activity in unheated primary infected milk was indeed caused by an interferon. Interferon excretion in primary infected milk was investigated using a series of milk samples from three cows. Maximum interferon titres were found after 24 h, coinciding with or shortly after the first virus excretion peak. The results are discussed with particular reference to the use of primary infected milk in studies of the thermal inactivation of FMD-virus.     

Aspects of heat inactivation of foot-and-mouth disease virus in milk from intramammarily infected susceptible cows.

In skim milk obtained from susceptible cows after intramammary and intravenous inoculation (primary infected milk), foot-and-mouth disease (FMD) virus type O1 was slower inactivated by heat treatment than virus that had been added to pre-exposure skim milk. Residual virus infectivity in heated primary infected milk was more efficiently detected in bovine thyroid cell cultures than in secondary pig kidney (PK2) cell cultures. Untreated primary infected milk was found to inhibit both FMD-virus and vesicular stomatitis virus plaque formation in PK2 cells, suggesting the presence of interferon. The results of further tests confirmed that the interfering activity in unheated primary infected milk was indeed caused by an interferon. Interferon excretion in primary infected milk was investigated using a series of milk samples from three cows. Maximum interferon titres were found after 24 h, coinciding with or shortly after the first virus excretion peak. The results are discussed with particular reference to the use of primary infected milk in studies of the thermal inactivation of FMD-virus.     

Prospects,including time-frames,for improved foot and mouth disease vaccines

Inactivated foot and mouth disease (FMD) vaccines have been used successfully as part of eradication programmes. However, there are a number of concerns with the use of such vaccines and the recent outbreaks of FMD in disease-free countries have increased the need for improved FMD control strategies. To address this requirement, new generation FMD vaccines are being developed. Currently, one of the most promising of these vaccine candidates utilises an empty viral capsid subunit delivered to animals by a live virus vector. This candidate, a replication-defective recombinant human adenovirus containing the capsid and 3C proteinase coding regions of FMD virus (FMDV), induces an FMDV-specific neutralising antibody response in inoculated animals. Upon challenge with a virulent animal-passaged homologous virus, swine and cattle vaccinated with this recombinant adenovirus are protected from clinical signs of FMD as well as from FMDV replication. One inoculation of a high dose of this vaccine candidate protected swine from challenge as early as seven days after vaccination.     

The effect of vaccination regimen on the transfer of foot and mouth disease antibodies from the sow to her piglets

Four groups of pregnant sows were inoculated with type O1 foot and mouth disease (FMD) oil emulsion vaccine at various times before farrowing and samples of the sow's serum, colostrum and milk, and piglet's serum, collected during the first week after farrowing, were analysed for FMD virus neutralizing activity. No FMD neutralizing antibodies were detectable in the piglets serum at birth but they were present 1.5 h after suckling and peak titres were reached 1-3 days later. There was no significant difference between the antibody titres of colostrum samples collected from different teats at farrowing. However, similar samples collected 3 days later showed significant (P less than 0.005) fore to hind variation. The principal FMD virus neutralizing antibody class present in the sow's serum at farrowing and in their 3-day-old piglets was governed by the inoculation schedule employed. When the last vaccinations were given congruent to 30 days before farrowing (dbf) the predominant FMD virus neutralizing class was IgG. However, when the sows were vaccinated only congruent to 12 dbf the predominant class was IgM. A significant correlation was observed between the sow's serum titres and colostrum titres at farrowing (r = 0.90), and also between sows colostrum titres at farrowing and their 3-day-old piglets serum titres (r = 0.99).     

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.     

The effect of vaccination regimen on the transfer of foot and mouth disease antibodies from the sow to her piglets.

Four groups of pregnant sows were inoculated with type O1 foot and mouth disease (FMD) oil emulsion vaccine at various times before farrowing and samples of the sow''s serum, colostrum and milk, and piglet''s serum, collected during the first week after farrowing, were analysed for FMD virus neutralizing activity. No FMD neutralizing antibodies were detectable in the piglets serum at birth but they were present 1.5 h after suckling and peak titres were reached 1-3 days later. There was no significant difference between the antibody titres of colostrum samples collected from different teats at farrowing. However, similar samples collected 3 days later showed significant (P less than 0.005) fore to hind variation. The principal FMD virus neutralizing antibody class present in the sow''s serum at farrowing and in their 3-day-old piglets was governed by the inoculation schedule employed. When the last vaccinations were given congruent to 30 days before farrowing (dbf) the predominant FMD virus neutralizing class was IgG. However, when the sows were vaccinated only congruent to 12 dbf the predominant class was IgM. A significant correlation was observed between the sow''s serum titres and colostrum titres at farrowing (r = 0.90), and also between sows colostrum titres at farrowing and their 3-day-old piglets serum titres (r = 0.99).     

Effect of emergency FMD vaccine antigen payload on protection, sub-clinical infection and persistence following direct contact challenge of cattle

Previous work, in sheep vaccinated with emergency foot-and-mouth disease (FMD) vaccine, indicated the benefit of increasing the antigen payload in inhibiting local virus replication and consequently persistence following an indirect aerosol challenge with a virus homologous to the vaccine strain. The work presented here investigates this possibility further using cattle and a more severe semi-heterologous direct contact challenge. The quantitative dynamics of virus replication and excretion in both vaccinated and non-vaccinated cattle following challenge are examined. Two experiments were carried out each involving 20 vaccinated and 5 non-vaccinated cattle. An O(1) Manisa vaccine (18 PD(50)) was used for the first, previously reported experiment [Cox SJ, Voyce C, Parida S, Reid SM, Hamblin PA, Paton DJ, et al. Protection against direct contact challenge following emergency FMD vaccination of cattle and the effect on virus excretion from the oropharynx. Vaccine 2005;23:1106-13]. The same vaccine was used for the second experiment described in this paper except the antigen payload was increased 10-fold per bovine dose, resulting in significantly higher FMD virus neutralising antibody titres prior to challenge. Twenty-one days post-vaccination the cattle received a 5-day direct contact challenge with FMD virus from five further non-vaccinated cattle infected 24h earlier with O UKG 34/2001. All vaccinated cattle regardless of antigen payload were protected against clinical disease. Sub-clinical oropharyngeal infection was detected in animals from both experiments but the level of virus replication shortly after direct contact challenge was significantly reduced in vaccinated animals. Cattle immunised with the 10-fold antigen payload cleared the virus more readily and consequently at 28 days post-challenge fewer animals were persistently infected compared to the single strength vaccine. Following a severe challenge, the results from both experiments show that use of emergency vaccine can prevent or decrease local virus replication and thereby dramatically reduce the amount of virus released into the environment, particularly during the early post-exposure period. Additionally, increasing the antigen payload of the vaccine may reduce sub-clinical infection, leading to fewer persistently infected virus carrier animals.     

Vaccination against foot and mouth disease reduces virus transmission in groups of calves

The aim of vaccination during an epidemic of foot and mouth disease (FMD) is not to induce clinical protection, but to reduce virus transmission. Since no quantitative data were available on the effectiveness of vaccination in cattle, we investigated whether a single vaccination against FMD could reduce virus transmission in groups of calves by estimating the reproduction ratio R, i.e. the average number of secondary cases caused by one infectious animal in a susceptible population. We performed two experiments with six groups of either four vaccinated or four non-vaccinated calves each. Vaccination was carried out with O(1) Manisa vaccine. Two weeks after vaccination, two calves per group were inoculated intra-nasally with FMDV field isolate O/NET 2001. The two other calves were contact-exposed to the inoculated calves. Contact infections were observed by clinical inspection, virus isolation and RT-PCR on heparinised blood, oro-pharyngeal fluid and probang samples and antibody response to non-structural proteins. In all six non-vaccinated groups, transmission to contact-exposed calves was recorded; in the vaccinated groups, virus transmission was observed to one contact-exposed calf. In the non-vaccinated groups R(c) was 2.52 and significantly above 1, whereas in the vaccinated groups R(v)=0.18 and significantly below 1, indicating that vaccination may successfully be applied as additional intervention tool to reduce virus transmission in a future epidemic of FMD.     

Protective immunity against foot-and-mouth disease virus induced by a recombinant vaccinia virus

We report the construction of a recombinant vaccinia virus expressing the precursor for the four structural proteins of FMD virus (FMDV) (P1) strain C3Arg85 using a procedure for isolation of recombinant vaccinia viruses based solely on plaque formation. Adult mice vaccinated with this recombinant vaccinia virus elicited high titers of neutralizing antibodies against both the homologous FMDV and vaccinia virus, measured by neutralization assays. Liquid phase blocking sandwich enzyme-linked immunosorbent assays (ELISAs) using whole virus as antigen showed high total antibody titers against homologous FMDV, similar to those induced by the conventional inactivated vaccine. When ELISAs were carried out with heterologous strains A79 or O1Caseros as antigens, sera from animals vaccinated with the recombinant virus cross-reacted. Mice boosted once with the recombinant vaccinia virus were protected against challenge with infectious homologous virus. These results indicate that recombinant vaccinia viruses are efficient immunogens against FMDV when used as a live vaccine in a mouse model.     

A foot-and-mouth disease SAT2 vaccine protects swine against experimental challenge with a homologous virus strain,irrespective of mild pathogenicity in this species

FMDV serotype SAT2 is most frequently associated with outbreaks in ruminants. However, the risk of it spreading from cattle to pigs cannot be excluded. To assess the efficacy of an SAT2-type FMD inactivated vaccine against homologous challenge in pigs, a suitable challenge strain adapted to pigs was produced. After two passages in two pigs each, a FMDV stock of SAT2 challenge strain was produced. This material was used to infect two groups of five pigs. The first group being vaccinated 28 days before challenge and the other one left as an unvaccinated control. Clinical signs were recorded, virus shedding was assessed on mouth swabs, and neutralising antibody titres were determined. At least 80% of the vaccinated pigs were protected against clinical disease. Furthermore, no virus shedding was observed in any of the vaccinated pigs. This study shows that experimentally inoculated pigs can become infected with a SAT2 serotype. Furthermore, vaccination offers protection against generalisation and viral excretion, confirming the potential of vaccination as an important tool in the control of FMD in pigs.     

Isotype-specific antibody responses to foot-and-mouth disease virus in sera and secretions of "carrier' and "non-carrier' cattle.

Isotype-specific antibody responses to foot-and-mouth disease virus (FMDV) were measured in the sera and upper respiratory tract secretions of vaccinated and susceptible cattle challenged with FMDV by direct contact or by intranasal inoculation. A comparison was made between cattle that eliminated FMDV and those that developed and maintained a persistent infection. Serological and mucosal antibody responses were detected in all animals after challenge. IgA and IgM were detected before the development of IgG1 and IgG2 responses. IgM was not detected in vaccinated cattle. Challenge with FMDV elicited a prolonged biphasic secretory antibody response in FMDV "carrier'' animals only. The response was detected as FMDV-specific IgA in both mucosal secretions and serum samples, which gained statistical significance (P < 0.05) by 5 weeks after challenge. This observation could represent the basis of a test to differentiate vaccinated and/or recovered convalescent cattle from FMDV "carriers''.     

Comparison of vaccination with rhesus CMV (RhCMV) soluble gB with a RhCMV replication-defective virus deleted for MHC class I immune evasion genes in a RhCMV challenge model

A human cytomegalovirus (HCMV) vaccine to prevent infection and/or reduce disease associated with congenital infection or visceral disease in transplant recipients is a high priority, but has remained elusive. We created a disabled infectious single cycle rhesus CMV (RhCMV) deleted for glycoprotein L (gL) and the MHC class I immune evasion genes Rh178 and Rh182-189, and restored its epithelial cell tropism by inserting the Rh128-131A genes. The resulting virus, RhCMVRΔgL/178/182–189, was used to vaccinate rhesus monkeys intramuscularly and was compared with vaccination of animals with soluble RhCMV glycoprotein B (gB) in alum/monophosphoryl lipid A or with PBS as a control. At 4?weeks after the second vaccination, an increased frequency of RhCMV-specific CD8 T cells was detected in animals vaccinated with the RhCMVRΔgL/178/182–189 vaccine compared to animals vaccinated with soluble gB. In contrast, monkeys vaccinated with soluble gB had 20-fold higher gB antibody titers than animals vaccinated with RhCMVRΔgL/178/182–189. Titers of neutralizing antibody to RhCMV infection of fibroblasts were higher in animals vaccinated with gB compared with RhCMVRΔgL/178/182–189. Following vaccination, monkeys were challenged subcutaneously with RhCMV UCD59, a low passage virus propagated in monkey kidney epithelial cells. All animals became infected after challenge; however, the frequency of RhCMV detection in the blood was reduced in monkeys vaccinated with soluble gB compared with those vaccinated with RhCMVRΔgL/178/182–189. The frequency of challenge virus shedding in the urine and saliva and the RhCMV copy number shed at these sites was not different in animals vaccinated with RhCMVRΔgL/178/182–189 or soluble gB compared with those that received PBS before challenge. Although the RhCMVRΔgL/178/182–189 vaccine was superior in inducing cellular immunity to RhCMV, it induced lower titers of neutralizing antibody and antibody to gB than the soluble gB vaccine; after challenge, animals vaccinated with soluble gB had a lower frequency of virus detection in the blood than those vaccinated with RhCMVRΔgL/178/182–189.     

Growth of foot-and-mouth disease virus in the upper respiratory tract of non-immunized, vaccinated, and recovered cattle after intranasal inoculation.

Non-immunized, vaccinated, and recovered cattle were inoculated intranasally with various doses of foot-and-mouth disease virus. Samples of oesophageal-pharyngeal (OP) fluid were taken periodically for up to 7 days after inoculation and virus titres of these samples were plotted as pharyngeal virus growth curves. In non-immunized cattle, the length of the lag period and of the growth period were inversely proportional to the dose of virus given. Maximum titres were observed when clinical signs were first detected. Three of the 10 cattle studied had virus growth rates that were lower than rates of others given the same dose of virus, and clinical signs appeared later than expected in these three cattle. Cattle vaccinated with an inactivated virus oil-adjuvant vaccine had pharyngeal virus growth curves that were similar to those obtained from non-immunized cattle for 30 h. after inoculation. Titres of virus in OP fluid samples taken 2-7 days after inoculation were substantially lower in cattle with a high pre-exposure serum mouse protection index than titres from partly-immunized or non-immunized cattle. Nine of 14 cattle had detectable but reduced virus growth after intranasal inoculation with homologous virus. Five recovered cattle inoculated with heterologous virus reacted similarly to non-immunized animals.     

Efficient priming against classical swine fever with a safe glycoprotein E2 expressing Orf virus recombinant (ORFV VrV-E2)

An increasing demand in livestock animal husbandry for intervention or emergency vaccination strategies requires a rapid onset of protection linked to prevention of infectious agent spread. Using the new recombinant parapoxvirus (PPV) Orf virus (ORFV) as a vaccine expressing the CSFV E2 glycoprotein we demonstrate that a single intra-muscular application confers solid protection. In the prime only concept, multi-site application of the vector vaccine turned out to be superior to single-site application as no pyrexia occurred after virulent CSFV challenge and CSFV neutralizing serum antibodies regularly were detectable before challenge. Vector virus vaccinated swine were able to cope with the lymphocyte and in particular B-cell depression in peripheral blood after challenge showing no clinical signs and no viremia. Early after challenge CSFV-specific IFN gamma production (IFN-gamma) and high neutralizing serum antibody titers clearly differentiated na?ve from vaccinated and protected animals. After CSFV challenge neutralizing serum antibodies titers in vector vaccinated swine were significantly higher than those in sera from live attenuated vaccine primed animals. Horizontal challenge virus transmission was prevented under strict sentinel isolation before mingling but not in next-door stables separated by a wooden barrier at the day of challenge. The presence of CSFV-specific pre-challenge serum antibodies although in low titers is a good prognostic parameter for solid protection after ORFV vector vaccination even when a significant CSFV-specific IFN-gamma production was not detectable before challenge. A heterologous prime-boost regimen as a combination of prime with baculovirus-expressed glycoprotein E2 followed by boost with the parapoxvirus vector turned out to be a better immune stimulant than a homologous prime/boost with the modified live CSFV vaccine. A similar beneficial effect became evident when the challenge infection mimicked the booster vaccination after a single PPV vector prime.     

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.     

Use of ENABL® adjuvant to increase the potency of an adenovirus-vectored foot-and-mouth disease virus serotype A subunit vaccine

A foot-and-mouth disease (FMD) recombinant subunit vaccine formulated with a lipid/polymer adjuvant was evaluated in two vaccine efficacy challenge studies in steers. The vaccine active ingredient is a replication-deficient human adenovirus serotype 5 vector encoding the FMD virus (FMDV) A24/Cruzeiro/BRA/55 capsid (AdtA24). In the first study, AdtA24 formulated in ENABL® adjuvant was compared to a fourfold higher dose of AdtA24 without adjuvant. Steers vaccinated with AdtA24 + ENABL® adjuvant developed a significantly higher virus neutralizing test (VNT) antibody titer and an improved clinical response following FMDV A24/Cruzeiro/BRA/55 intradermal lingual challenge at 14 days post-vaccination (dpv) than steers vaccinated with the active ingredient alone. In the second study, vaccination with AdtA24 formulated in ENABL® at the same dose used in the first study, followed by FMDV A24/Cruzeiro/BRA/55 challenge on 7 or 14 dpv, prevented clinical FMD in all steers and conferred 90% protection against viremia. In addition, post-challenge FMDV titers in nasal samples from vaccinated steers compared to unvaccinated steers were significantly reduced. In both studies, none of the AdtA24 vaccinated steers developed antibodies to the FMDV non-structural proteins prior to challenge with FMDV, indicative of the capacity to differentiate infected from vaccinated animals (DIVA). These results demonstrate that administration of AdtA24 formulated in ENABL® adjuvant lowered the protective dose and prevented clinical FMD following exposure of vaccinated steers to virulent FMDV at 7 or 14 dpv.     

Quantification of foot and mouth disease virus excretion and transmission within groups of lambs with and without vaccination

Sheep are well known to be susceptible for foot and mouth disease virus (FMDV), but it is unknown whether the infection can spread and persist in a sheep population. We therefore quantified virus transmission by performing experiments with FMD virus strain O/NET/2001 in groups of lambs. We used six groups of four lambs each, in which half of each group was inoculated and the other half was contact-exposed. To quantify the effectiveness of a single vaccination we also included six groups of four lambs each, vaccinated with O Manisa vaccine, 14 days prior to inoculation. Oropharyngeal fluid was obtained with a swab (OPF-swab), and blood samples were collected daily to determine virus excretion and serological response. We calculated the transmission rate beta (the number of new infections per day per infectious animal), and the reproduction ratio R (the number of secondary infections caused by one infectious individual). The mean daily virus excretion and the number of days the lambs excreted virus in the OPF differed significantly between vaccinated and non-vaccinated lambs. The transmission rate beta in the unvaccinated groups was 0.105 (95% confidence limit 0.044; 0.253) per day. The duration of the infectious period (T) was 21.11 (95% confidence limit 10.6; 42.1) days. With the final size of infection we estimated the reproduction ratio R in the non-vaccinated groups to be 1.14 (0.3; 3.3), and in vaccinated groups 0.22 (0.01; 1.78). Virus transmission as quantified by the final size did not differ statistically between the vaccinated and the non-vaccinated groups of lambs. In conclusion, FMDV seems able to persist in a sheep population, although the reproduction ratio was only slightly larger than one. As a consequence, vaccination might only have a small effect on transmission, in spite of the fact that virus excretion and duration of virus excretion is significantly reduced after vaccination.     

Chemokine CCL20 plasmid improves protective efficacy of the Montanide ISA™ 206 adjuvanted foot-and-mouth disease vaccine in mice model

This study aimed to investigate the chemokine CCL20, a macrophage inflammatory protein-3 alpha, for adjuvant potential in inactivated foot-and-mouth disease (FMD) vaccine. Groups of mice were injected intramuscularly with either murine CCL20 DNA or CCL20 protein two days ahead of the immunization with Montanide ISA206 adjuvanted inactivated FMD vaccine and humoral and cellular immune responses were measured in post-vaccinal sera. We demonstrated that the mice immunized with CCL20 plasmid plus FMD vaccine showed earlier and significantly (p < 0.05) higher neutralizing antibody responses compared to the mice vaccinated with CCL20 protein plus FMD vaccine. In fact, CCL20 as a protein did not show any adjuvant effect and the immune responses induced in this group were comparable to that of the mice vaccinated with FMD vaccine alone. All the vaccination groups showed serum IgG1 and IgG2 antibody responses; however, the mice vaccinated with CCL20 plasmid plus FMD vaccine showed significantly (p < 0.05) higher IgG1 and IgG2 responses and the responses remained high at all-time points post vaccination, although not always statistically significant. Upon restimulation of the vaccinated splenocytes with the inactivated FMD viral antigen, significantly (p < 0.05) higher IFN-γ and IL-2 levels in culture supernatants were found in animals vaccinated with the CCL20 plasmid plus FMD vaccine, which is indicative of the T_H1 type of cellular immunity. On challenge with the homologous FMD virus on 28th day post immunization, CCL20 plasmid plus FMD vaccine showed complete protection (100%) while animals immunized with CCL20 protein plus FMD vaccine or FMD vaccine alone showed 66% protection. In summary, we show that prior injection of CCL20 plasmid improved protective efficacy of the inactivated FMD vaccine and thus offers a valuable strategy to modulate the efficacy and polarization of specific immunity against inactivated vaccines.