Vaccinology of classical swine fever: from lab to field

There are two types of classical swine fever vaccines available: the classical live and the recently developed E2 subunit vaccines. The live Chinese strain vaccine is the most widely used. After a single vaccination, it confers solid immunity within a few days that appears to persist lifelong. The E2 subunit vaccine induces immunity from approximately 10-14 days after a single vaccination. The immunity may persist for more than a year, but is then not complete. The Chinese strain vaccine may establish a strong herd immunity 1-2 weeks earlier than the E2 vaccine. The ability of the Chinese vaccine strain to prevent congenital infection has not been reported, but the E2 subunit vaccine does not induce complete protection against congenital infection. Immunological mechanisms that underlie the protective immunity are still to be elucidated. Both types of vaccine are considered to be safe. A great advantage of the E2 subunit vaccine is that it allows differentiation of infected pigs from vaccinated pigs and is referred to as a DIVA vaccine. However, the companion diagnostic E(rns) ELISA to actually make that differentiation should be improved. Many approaches to develop novel vaccines have been described, but none of these is likely to result in a new DIVA vaccine reaching the market in the next 5-10 years. Countries where classical swine fever is endemic can best control the infection by systematic vaccination campaigns, accompanied by the normal diagnostic procedures and control measures. Oral vaccination of wild boar may contribute to lowering the incidence of classical swine fever, and consequently diminishing the threat of virus introduction into domestic pigs. Free countries should not vaccinate and should be highly alert to rapidly diagnose any new outbreak. Once a new introduction of classical swine fever virus in dense pig areas has been confirmed, an emergency vaccination programme should be immediately instituted, for maximum benefit. The question is whether the time is ripe to seriously consider global eradication of classical swine fever virus.     

The effects of challenge on the humoral and cellular immune responses in pseudorabies vaccinated swine.

The effects of challenge exposure on the humoral and cellular immune responses in pseudorabies vaccinated swine were studied in 84 barrows. The pigs were divided into seven groups and challenge exposed to a virulent strain of pseudorabies virus on months 1, 3, 5, 8, 10, 12 and 14 after vaccination. The pigs were vaccinated with commercial attenuated and inactivated pseudorabies virus vaccines. The protection conferred by vaccination was equally effective with both types of vaccines. The levels of cellular and humoral immunity after challenge exposure in pigs vaccinated with either type of vaccine were similar. The cell-mediated immune response can be effectively used for the early detection of pigs exposed to pseudorabies virus. Virus isolation attempts from the brain and spleen in most of the vaccinated pigs were unsuccessful.     

Protective efficacy of a Classical swine fever virus C-strain deletion mutant and ability to differentiate infected from vaccinated animals

Classical swine fever (CSF) continues to be the most economically damaging pig disease in the world. The disease can be effectively controlled by vaccination with the live C-strain vaccine. This vaccine, however, does not enable the serological differentiation between infected and vaccinated animals (DIVA) and its use can therefore impose severe trade restrictions. CSF-specific diagnostic ELISAs detect antibodies directed against the conserved and immunodominant A domain of the E2 structural glycoprotein. We previously reported the production of a C-strain virus in which the immunodominant TAVSPTTLR epitope of the A domain is stably mutated with the aim to render the virus suitable as a DIVA vaccine. We here report that a single vaccination with this vaccine virus protected pigs from a lethal challenge dose of the highly virulent Brescia strain. Analysis of the sera, however, demonstrated that a commercially available E2 ELISA was unsuitable as an accompanying DIVA test.     

Antibody responses to inactivated vaccines and natural infection in cattle using bovine viral diarrhoea virus ELISA kits: Assessment of potential to differentiate infected and vaccinated animals

Bovine viral diarrhoea virus (BVDV) is one of the most common and economically important viral infections of cattle. As vaccination is common in most European countries, differentiation between infected and vaccinated animals is one of the key challenges facing BVDV eradication campaigns. This study was designed to compare the ability of commercial ELISA kits to differentiate antibodies generated following vaccination with four different commercial inactivated BVDV vaccines from antibodies generated following challenge with virulent BVDV. Although none of the tested vaccine–ELISA combinations was able to differentiate an infected from a vaccinated animal (DIVA) at the individual animal level, p80 blocking ELISAs, in combination with inactivated BVDV vaccines, may have some value under certain circumstances at herd level. In most cases, antibody responses to BVDV vaccines cannot be clearly distinguished from responses seen in the early phase of natural infection. No commercial BVD vaccine showed true marker qualities for DIVA using p80 blocking ELISAs.     

Pseudorabies virus eradication by area‐wide vaccination is feasible

Summary

This article reviews the rationale for using marker vaccines and companion diagnostic tests in the eradication of pseudorabies virus (PRV). Recent advances in vaccinology and epidemiology indicate that, despite the inability to induce complete immunity, vaccination is a useful tool in the battle against PRV. This review focuses on the effectiveness of vaccination under field conditions and on herd, management and regional factors that are associated with PRV introduction or transmission.     

禽流感免疫预防的探讨

近年来,在禽流感新型疫苗研究上取得了一定进展,但是目前国内外禽流感免疫仍主要使用灭活疫苗.大量的研究和防控实践表明,有效的禽流感疫苗接种可预防家禽发病,阻断病毒的传播,即使免疫禽受到强毒攻击感染后病毒排出量也显著减少,这就减少了病毒发生自然突变的机会,降低了疫病向禽群和人传播的风险;疫苗的正确应用可在控制禽流感中发挥重要作用.因此,应根据不同地区总体防控策略和具体疫情状况,正确评估禽流感免疫预防作用.在禽流感发生的高风险地区,全面预防接种是防控禽流感的关键技术措施,具有至关重要的作用.     

Vaccination against classical swine fever virus: limitations and new strategies

The most widely used vaccines for the control of classical swine fever (CSF) in countries where it is endemic are live attenuated virus strains, which are highly efficacious, inducing virtually complete protection against challenge with pathogenic virus. In the European Union (EU), the combination of prophylactic mass vaccination and culling of infected pigs in endemic regions has made it possible to almost eradicate the disease. However, it is not possible to discriminate between infected and vaccinated animals, thus hampering disease control measures that rely on serology. Therefore, vaccination was banned at the end of 1990 before the internal common market was established in the EU. Vaccination is allowed only in severe emergencies. In addition, there are strict restrictions on the international trade in pig products from countries using vaccination. To circumvent these problems, marker vaccines which allow differentiation of infected from vaccinated animals (DIVA) have been developed. There are several approaches, ranging from protective peptides, single expressed proteins, naked DNA and chimeric viruses. To date, two subunit vaccines based on the E2 glycoprotein are commercially available and have been tested extensively for their efficacy. The accompanying discriminatory tests are based on an ELISA detecting another viral glycoprotein, the E(rns). The subunit vaccines were found to be less efficacious than live attenuated vaccines. In addition, the currently available discriminatory tests do not provide high enough specificity and sensitivity. Although there is an urgent need for more advanced marker vaccines and better discriminatory tests, the development of new DIVA vaccines against CSF is hampered by the small market potential for these products.     

Avian influenza virus: prospects for prevention and control by vaccination

Although vaccination does not always prevent infection of avian influenza (AI) virus, the clear benefit of vaccination is in its ability to prevent disease and to reduce the amount of virus in circulation. Thus, judicious use of vaccination can be an important component of an AI control program. However, the long-term use of vaccination without eradication may result in the selection of the antigenically divergent strains, which compromises the value of vaccination. In this review, the effectiveness of currently available and future AI vaccines is discussed with suggestions for the ideal use of vaccination even with antigenic drift of the virus.     

Recombinant adenovirus expressing the haemagglutinin of peste des petits ruminants virus (PPRV) protects goats against challenge with pathogenic virus; a DIVA vaccine for PPR

Peste des petits ruminants virus (PPRV) is a morbillivirus that can cause severe disease in sheep and goats, characterised by pyrexia, pneumo-enteritis, and gastritis. The socio-economic burden of the disease is increasing in underdeveloped countries, with poor livestock keepers being affected the most. Current vaccines consist of cell-culture attenuated strains of PPRV, which induce a similar antibody profile to that induced by natural infection. Generation of a vaccine that enables differentiation of infected from vaccinated animals (DIVA) would benefit PPR control and eradication programmes, particularly in the later stages of an eradication campaign and for countries where the disease is not endemic. In order to create a vaccine that would enable infected animals to be distinguished from vaccinated ones (DIVA vaccine), we have evaluated the immunogenicity of recombinant fowlpox (FP) and replication-defective recombinant human adenovirus 5 (Ad), expressing PPRV F and H proteins, in goats. The Ad constructs induced higher levels of virus-specific and neutralising antibodies, and primed greater numbers of CD8⁺ T cells than the FP-vectored vaccines. Importantly, a single dose of Ad-H, with or without the addition of Ad expressing ovine granulocyte macrophage colony-stimulating factor and/or ovine interleukin-2, not only induced strong antibody and cell-mediated immunity but also completely protected goats against challenge with virulent PPRV, 4 months after vaccination. Replication-defective Ad-H therefore offers the possibility of an effective DIVA vaccine.     

Comparison of two real-time RT-PCR assays for differentiation of C-strain vaccinated from classical swine fever infected pigs and wild boars

Classical swine fever is one of the most important infectious diseases for the pig industry worldwide due to its economic impact. Vaccination is an effective means to control disease, however within the EU its regular use is banned owing to the inability to differentiate infected and vaccinated animals, the so called DIVA principle. This inability complicates monitoring of disease and stops international trade thereby limiting use of the vaccine in many regions. The C-strain vaccine is safe to use and gives good protection. It is licensed for emergency vaccination in the EU in event of an outbreak. Two genetic assays that can distinguish between wild type virus and C-strain vaccines have recently been developed. Here the results from a comparison of these two real-time RT-PCR assays in an interlaboratory exercise are presented. Both assays showed similar performance.     

猪口蹄疫O型合成肽疫苗及其主要特点

口蹄疫是当前严重危害我国养猪业的疾病之一,长期以来免疫接种是我国预防该病的重要措施。近年来研制出的猪口蹄疫O型合成肽疫苗以其免疫原性高、生物安全性好、可有效区分免疫动物和感染动物等优点引起了人们的高度重视。本文主要介绍了口蹄疫病毒、O型口蹄疫病毒的抗原位点和猪口蹄疫O型合成肽疫苗的研究概况及其主要特点,旨在为猪口蹄疫O型合成肽疫苗的推广应用提供参考。     

口蹄疫自然感染动物与免疫动物鉴别诊断研究进展

口蹄疫是偶蹄动物高度接触性传染病 ,能引起巨大经济损失。国际兽医局将其列为 A类动物传染病之首。除发达国家外 ,大多数发展中国家都采用注射疫苗的办法来控制该病的流行。因此如何区分感染动物和免疫动物是口蹄疫防制中迫切需要解决的问题。目前口蹄疫灭活疫苗的生产工艺可以将绝大部分的非结构蛋白除去 ,因而灭活疫苗免疫动物只能产生结构蛋白抗体 ,而感染动物能产生结构蛋白抗体 ,也能产生非结构蛋白抗体 ,因此 ,检测非结构蛋白抗体为鉴别口蹄疫感染动物与免疫动物提供了美好前景。文章从鉴别诊断的原理 ,非结构蛋白的免疫特性 ,鉴别诊断所面临的问题及解决方案 ,应用非结构蛋白作为鉴别诊断抗原的研究现状等方面进行了综述。     

Pseudorabies virus latency and reactivation in vaccinated swine

Latency and reactivation of pseudorabies virus in swine was studied. Thirty-one pigs were assigned to 5 groups and were given 1 of 4 vaccines; 10 remained unvaccinated controls. All pigs were then challenge exposed with a sublethal dose of virulent pseudorabies virus. One hundred one days after challenge exposure, all pigs were treated with dexamethasone to reactivate the virus. Virus-positive tonsil and nasal mucus isolates were recovered from 29 of the 31 pigs over a 12-day period. Frequency and duration of virus-positivity were significantly (P less than 0.05) and consistently lower among vaccinated pigs than among the unvaccinated controls. It was concluded that vaccination before challenge exposure had little or no effect on the rate of establishment of virus latency, but that vaccination reduced shedding after subsequent reactivation of the virus.     

基于新城疫病毒V蛋白鉴别鸡群感染与免疫的间接ELISA方法的建立

本研究以新城疫病毒(NDV)V蛋白羧基端结构域(Vc)的重组蛋白为包被抗原,建立了用于检测NDV V蛋白抗体的间接ELISA方法,并采用该方法检测了鸡群免疫或接毒后血清中的V蛋白抗体水平。结果显示:两组不同NDV灭活疫苗组在免疫后的3周内检测结果均为阴性;两组灭活疫苗免疫3周后再人工感染NDV强毒的鸡群,攻毒后第7、14和21 d,NDV阳性率分别为60%、80%、70%和50%、80%、70%;两组不同的NDV弱毒疫苗免疫组鸡群,仅在免疫后第21 d阳性率分别为20%和10%。以上结果表明,NDV疫苗免疫组与强毒感染组的V蛋白抗体阳性率存在明显差异,本方法可在群体水平上区分新城疫疫苗免疫与强毒感染鸡群,为NDV血清学诊断和流行病学调查提供了一种新的检测手段。     

Genetic differentiation of infected from vaccinated animals after implementation of an emergency vaccination strategy against classical swine fever in wild boar

Oral emergency vaccination against classical swine fever is a powerful tool to control disease outbreaks among European wild boar and thus to safeguard domestic pigs in affected regions. In the past, when virus detection was mainly done using virus isolation in cell culture or antigen enzyme-linked immunosorbent assays, modified live vaccine strains like C-strain "Riems", were barely detectable after oral vaccination campaigns. Nowadays, the use of highly sensitive molecular techniques has given rise to an increase in vaccine virus detections. This was also the case during the 2009 outbreak among German wild boar and the subsequent vaccination campaigns. To guarantee a rapid differentiation of truly infected from C-strain vaccinated animals, a combination of differentiating multiplex rRT-PCR assays with partial sequencing was implemented. Here, we report on the rational and use of this approach and the lessons learned during execution. It was shown that positive results in the recently developed vaccine strain (genotype) specific rRT-PCR assay can be taken as almost evidentiary whereas negative results should be confirmed by partial sequencing. Thus, combination of multiplex rRT-PCR assays as a first line differentiation with partial sequencing can be recommended for a genetic DIVA strategy in areas with oral vaccination against classical swine fever in wild boars.     

Effect of various vaccination procedures on shedding, latency, and reactivation of attenuated and virulent pseudorabies virus in swine.

Various procedures of vaccination for pseudorabies were compared for their effects on shedding, latency, and reactivation of attenuated and virulent pseudorabies virus. The study included 6 groups: group 1 (10 swine neither vaccinated nor challenge-exposed), group 2 (20 swine not vaccinated, but challenge-exposed), and groups 3 through 6 (10 swine/group, all vaccinated and challenge-exposed). Swine were vaccinated with killed virus IM (group 3) or intranasally (group 4), or with live virus IM (group 5) or intranasally (group 6). The chronologic order of treatments was as follows: vaccination (week 0), challenge of immunity by oronasal exposure to virulent virus (week 4), biopsy of tonsillar tissue (week 12), treatment with dexamethasone in an attempt to reactivate latent virus (week 15), and necropsy (week 21). Vaccination IM with killed or live virus and vaccination intranasally with live virus mitigated clinical signs and markedly reduced the magnitude and duration of virus shedding after challenge exposure. Abatement of signs and shedding was most pronounced for swine that had been vaccinated intranasally with live virus. All swine, except 4 from group 2 and 1 from group 4, survived challenge exposure. Only vaccination intranasally with live virus was effective in reducing the magnitude and duration of virus shedding after virus reactivation. Vaccination intranasally with killed virus was without measurable effect on immunity. Of the 55 swine that survived challenge exposure, 54 were shown subsequently to have latent infections by use of dexamethasone-induced virus reactivation, and 53 were shown to have latent infections by use of polymerase chain reaction (PCR) with trigeminal ganglia specimens collected at necropsy. Fewer swine were identified by PCR as having latent infections when other tissues were examined; 20 were identified by testing specimens of olfactory bulbs, 4 by testing tonsil specimens collected at necropsy, and 4 by testing tonsillar biopsy specimens. Eighteen of the 20 specimens of olfactory bulbs and 3 of the 4 tonsil specimens collected at necropsy in which virus was detected by PCR were from swine without detectable virus-neutralizing antibody at the time of challenge exposure. One pig that had been vaccinated intranasally with live virus shed vaccine virus from the nose and virulent virus from the pharynx concurrently after dexamethasone treatment. Evaluation of both viral populations for unique strain characteristics failed to provide evidence of virus recombination.     

Analysis of glycoprotein I (gI) negative and aberrant pseudorabies viral diagnostic isolates.

Glycoprotein I (gI) phenotypes and genotypes of 4 pseudorabies viral diagnostic isolates were evaluated by use of in vitro DNA amplification, monoclonal antibody binding, gI-specific serodiagnostic responses, and in vivo virulence approaches. Three viruses were avirulent and did not elicit gI-specific serologic responses, react with gI-specific monoclonal antibodies, or contain gI epitope-encoding DNA sequences. The fourth virus was virulent and did elicit a gI-specific serodiagnostic response. Compared with reference virulent pseudorabies viruses, however, the fourth isolate had reduced reactivity with a group of gI monoclonal antibodies and had a single nucleotide sequence substitution with a corresponding putative amino acid change in the epitopically dominant portion of the gI molecule. Presumably, the first 3 isolates represented diagnostic recoveries of viruses derived from gI-deleted modified-live pseudorabies viral vaccines, whereas the fourth isolate was a virulent but gI-aberrant wild-type virus. Thoroughly assessing the gI status of pseudorabies viral diagnostic isolates was considered to be essential in evaluating the epidemiologic importance of these viruses and in monitoring the validity of gI-based vaccine companion tests now used worldwide in pseudorabies control and eradication programs.     

Round table on control of Aujeszky's disease and vaccine development based on molecular biology.

A summary is given on the 4 topics which were discussed during the round table and which represent current knowledge on the molecular biology of Aujeszky's disease (pseudorabies) virus. They include a review on 1. the genome and gene products of the virus; 2. the viral genes associated with virulence; 3. the immunological role of the viral gene products and 4. studies intended to compare the efficacy of several commercially available vaccines and to establish a possible correlation between antibodies against individual structural viral proteins and degree of protection. It was concluded that gI deleted vaccines appear to be the best choice for use in intensive vaccination programmes towards eradication of Aujeszky's disease virus. However, there remains a need for development of more potent vaccines which induce strong humoral and cell mediated immune responses and afford complete protection, virological protection included. It is often observed that live vaccine strains which are completely avirulent lose much capacity to replicate and spread within the vaccinated animal. It is, however, not excluded that a certain degree of dissemination may be needed to be fully efficacious. Loss of virulence may thus be accompanied by too much loss of immunogenicity. An improved genetic stability of live vaccine strains when they are obtained by genetic manipulation, possibly justifies a more widespread dissemination of the vaccine strain in the body compared to that with conventionally developed strains or compared to what is presently allowed.     

Vaccinating chickens against avian influenza with fowlpox recombinants expressing the H7 haemagglutinin

OBJECTIVE: To evaluate the vaccine efficacy of a fowlpox virus recombinant expressing the H7 haemagglutinin of avian influenza virus in poultry. PROCEDURE: Specific-pathogen-free poultry were vaccinated with fowlpox recombinants expressing H7 or H1 haemagglutinins of influenza virus. Chickens were vaccinated at 2 or 7 days of age and challenged with virulent Australian avian influenza virus at 10 and 21 days later, respectively. Morbidity and mortality, body weight change and the development of immune responses to influenza haemagglutinin and nucleoprotein were recorded. RESULTS: Vaccination of poultry with fowlpox H7 avian influenza virus recombinants induced protective immune responses. All chickens vaccinated at 7 days of age and challenged 21 days later were protected from death. Few clinical signs of infection developed. In contrast, unvaccinated or chickens vaccinated with a non-recombinant fowlpox or a fowlpox expressing the H1 haemagglutinin of human influenza were highly susceptible to avian influenza. All those chickens died within 72 h of challenge. In younger chickens, vaccinated at 2 days of age and challenged 10 days later the protection was lower with 80% of chickens protected from death. Chickens surviving vaccination and challenge had high antibody responses to haemagglutinin and primary antibody responses to nucleoprotein suggesting that although vaccination protected substantially against disease it failed to completely prevent replication of the challenge avian influenza virus. CONCLUSION: Vaccination of chickens with fowlpox virus expressing the avian influenza H7 haemagglutinin provided good protection against experimental challenge with virulent avian influenza of H7 type. Although eradication will remain the method of first choice for control of avian influenza, in the circumstances of a continuing and widespread outbreak the availability of vaccines based upon fowlpox recombinants provides an additional method for disease control.     

Influenza vaccines and vaccination strategies in birds

Although it is well accepted that the present Asian H5N1 panzootic is predominantly an animal health problem, the human health implications and the risk of human pandemic have highlighted the need for more information and collaboration in the field of veterinary and human health. H5 and H7 avian influenza (AI) viruses have the unique property of becoming highly pathogenic (HPAI) during circulation in poultry. Therefore, the final objective of poultry vaccination against AI must be eradication of the virus and the disease. Actually, important differences exist in the control of avian and human influenza viruses. Firstly, unlike human vaccines that must be adapted to the circulating strain to provide adequate protection, avian influenza vaccination provides broader protection against HPAI viruses. Secondly, although clinical protection is the primary goal of human vaccines, poultry vaccination must also stop transmission to achieve efficient control of the disease. This paper addresses these differences by reviewing the current and future influenza vaccines and vaccination strategies in birds.