Hand, foot, and mouth disease (HFMD) is a highly contagious disease with no specific treatment. Since it is common in immunocompromised children under the age of 5, there is a need to develop a safe vaccine. Virus-like particles (VLPs) are similar structures to viruses with the lack of genetic material which makes them impossible to replicate and infect, and therefore have a high level of biological safety and are considered to have high value as vaccines. In this study, the insect virus expression system that is widely used for vaccine and drug production due to its high post-translational modification efficiency, was used to produce VLPs for Coxsackievirus type A6 and A10, which are recently reported to be the main causes of HFMD. For this purpose, the selection of promoters that can control the timing and intensity of expression of 3CD protein, which is essential for VLPs assembly but has been reported to be cytotoxic, was conducted to construct an optimal expression form for HFMD-VLP.
Foot-and-mouth disease (FMD), which affects cloven-hoofed animals, is economically important because of its highly contagious nature. FMD virus (FMDV), the causative agent of FMD, involves seven serotypes (O, A, Asia1, C, and SAT 1-3). Serotype Asia1 is unique to the Asian territory and is subdivided into nine genetic groups (G-I-IX) based on nucleotide variations in the VP1 sequence. Asia1 Shamir, the most representative Asia1 vaccine, is not highly protective against the Asia1/MOG/05 (G-V) lineage found in North Korea in 2007. Therefore, we investigated whether a chimeric virus strain (Asia1/MOG/Shamir), in which the VP4, VP2, and VP3 sequences of Asia1/MOG/05 were combined with the VP1 sequence of Asia1 Shamir, can simultaneously protect against both viruses. We determined the optimal viral growth conditions for the commercial utilization of this chimeric virus strain. Of the three types of cell culture media, the Cellvento medium resulted in the highest amount of antigen in the samples. The chimeric strain was proliferated in a small bioreactor to produce a test vaccine, and its immunogenicity was evaluated in pigs. The virus neutralization (VN) titer against the Asia1 Shamir virus was > 1/100 after the second immunization with the chimeric vaccine in pigs. In addition, a single dose of the test vaccine resulted in a VN titer of > 1/100 against the Asia1/MOG/05 strain. Taken together, our chimeric vaccine strain provided sufficient protection against the Asia1/MOG/05 and Asia1 Shamir viruses, suggesting its potential as a novel vaccine for both these strains.
The Republic of Korea has implemented an obligatory vaccination on major livestock breeds to prevent and eradicate nationwide foot and mouth disease (FMD) since the end of 2010. The government has executed massive serological survey to check the immune level of various herds after vaccination, and seropositive rates against FMD virus (FMDV) structural proteins can be measured to assess FMD immunity level. The purpose of this study is to investigate the FMDV serological level of every cattle breeding farm in the country and to determine whether there is a significant difference between groups classified by time, age, and management authority. A total of 5,781 serum samples was collected in 18 cattle breeding farms from 2020 to 2021, and the seropositive rates were measured using PrioCHECK FMDV Type O ELISA kit. Firstly, the cattle breeding farms were classified by which they are managed: the central government, the local government, and the private agency. Every management authority had a seropositive rate of 99.5% or higher. Secondly, the samples were divided into 6 to 12 months old, 12 to 24 months old, and 24 months or more. The 6 to 12 months old group in 2020 showed a significantly low seropositive rate of 98.1%, but it was improved by implementing the enhanced vaccination policy from 2021 to 100%. In summary, there are considerably high seropositive rates including all groups with time, age, and by which they are managed, which means the FMD vaccination in cattle breeding farms is well-managed.
Foot-and-mouth disease (FMD) is a highly contagious vesicular disease that affects cloven-hoofed animals, causing substantial economic losses to the livestock industry. The causative FMD virus (FMDV) comprises four structural proteins (VP1, VP2, VP3, and VP4) and several non-structural proteins. Among the capsid proteins, VP4 is the most conserved, making it an attractive target as a diagnostic and vaccine antigen, regardless of FMDV serotype. In this study, we attempted to express the VP4 protein N-terminally fused to a glutathione S-transferase (GST) tag in Escherichia coli. Whereas VP0 and VP2 proteins were expressed in the soluble fraction, we failed to detect VP4, even in the insoluble fraction. To investigate the effect of VP4 C-terminal amino acid residues on protein expression, we constructed three VP4 mutants fused to GST, among which the mutant in which the C-terminal 15 amino acid residues had been deleted showed the highest level of protein expression. Furthermore, protein expression was observed even in the mutant in which three amino acid residues (DKK) had been fused to the C terminus. However, unlike the other two mutants, the wild-type VP4 mutant was poorly expressed, thereby indicating that the C-terminal amino acid residues could play a pivotal role in determining expression of the VP4 protein in E. coli.
In the case of foot-and-mouth disease (FMD), there is a great deal of impact on the national economy due to the disposal of diseases, the cost of disease control such as vaccination, reduction of productivity, and restriction of international trade of livestock products. Therefore, appropriate diagnostic methods for sensitive, accurate and rapid identification of virus serotypes are continuously required in terms of early prevention of FMD. This study was conducted to confirm the feasibility of immuno-PCR diagnostic method for the more sensitive detection of Korean FMD virus (FMDV). We synthesized a partial FMD type A viral gene. Protein antigen, monoclonal and polyclonal antibodies of FMDV were cloned, expressed and purified and then magnetic particles were attached to polyclonal antibodies and and oligomers to monoclonal antibodies for the immnuno-PCR. We confirmed the antigen-antibody and oligomer reaction using ELISA, Western blot, and real-time PCR. These results show that Korean FMDV can be detected by using difference of Ct values between positive group and negative group using immuno-PCR.. The results of this study also suggest that this technique will be the basis of the diagnosis method to detect Korean FMDV more sensitively in the future.
A cell line of bovine origin was immortalized to isolate foot-and-mouth disease virus (FMDV). The immortalization was performed by infection of bovine primary epithelial cells with a recombinant retrovirus that overexpressed the human telomerase (hTERT), after primary culture of fetal bovine kidney tissue and removal of fibroblasts. After cloning the immor- talized cell line into single cells, the cloned cell lines were named JNUBK-1, JNUBK-2, JNUBK-3 and JNUBK-4, according to their characteristics. To confirm the epithelial phenotype of the cell lines JNUBK-3 and JNUBK-4, which showed stable proliferation capability over 35 generations after immortalization, the expression of cytokeratin and fibronectin was measured. Finally, the FMDV titer in the JNUBK-3 and JNUBK-4 cell lines was measured and was 800∼2,000 times higher than that of the currently used cell line IRBS-2. In conclusion, more sensitive isolation and production of FMDV became possible through the use of the immortalized JNUBK-3 and JNUBK-4 cell lines.
구제역 바이러스는 소나 돼지 등의 우제류 가축에 경구 또는 흡입 노출 등의 다양한 노출 경로를 통해 심각한 감염성을 유발하는 생물학적 유해인자로 국내 축산업 운영에 있어 막대한 경제적 손실을 초래하는 병원균들 중 하나다. 구제역 발병에 대한 사전적 예방 관리를 도모할 수 있는 조치 방안 중 하나는 공기를 통해 전파되는 구제역 바이러스 확산에 대해 수학적 모델 적용을 통해 예측된 분석 결과에 따라 설정될 수 있다. 대기 확산 모델들은 일반적으로 구제역 바이러스의 주요 전파 경로인 경구 및 접촉 감염의 노출 시나리오를 예측할 수는 없으나, 상대적으로 예측도가 높은 대기 확산 모델의 경우 공기 전파에 대한 구제역 바이러스의 위해성 관리 방안을 결정하는 데 주요 역할을 담당할 수 있는 유용가능한 수단으로 여러 나라에서 활용되어 왔다. 구제역 바이러스 전파를 억제하기 위한 엄격한 관리 방안 중 다량의 가축 살처분 방법은 축산 농가들에게 심각한 경제적 손실을 초래하여 심각한 경우 새로운 자립의 여건마저도 상실시킬 수도 있다. 반면 낮은 수준의 대응 방안은 향후 구제역 발생에 따른 추가적 피해를 근원적으로 억제할 수 없는 역학적 한계에 봉착할 수 있다. 본 연구는 구제역 바이러스의 기본 특성 및 발생 모델 적용에 따른 공기 중 전파의 감염 위험성 평가 관련 선행 문헌들을 고찰하였다. 또한 1970년대 이후 구제역 바이러스의 공기 전파 경로를 예측하기 위해 여러 나라에서 실제 활용된 다양한 대기 확산 모델들의 적용 사례 및 장/단점을 비교 분석하였다.